Chiba University, Japan
This is my first research internship, and I had the wonderful opportunity to spend two months (Aug 1–Sep 26, 2025) at NIBB in the Cellular Dynamics Laboratory.
From the moment I arrived, I was warmly welcomed by Prof. Ueda, Dr. Kanazawa, Ms. Okubo, and all lab members. The lab environment is highly stimulating, with access to advanced equipment and supportive colleagues. Even it was a short period, I had improvement in my technical skills and deeper understanding of experimental reasoning.
During my internship, I carried out oil body observation in Marchantia polymorpha seedlings using confocal microscopy, where I was taught how to get clearly visualize structures and capture images for analysis. I took part on experiments, including protein extraction, quantification, SDS-PAGE, transfer, and antibody detection, which provided me with confidence in handling the entire process. In addition, I performed Co-immunoprecipitation (Co-IP) combined with immunoblotting to explore potential interactors of GFP-MpSYP12B under estradiol and paclitaxel (taxol) induction of MpERF13. While the Co-IP results varied in signal strength and background, adjustments to buffer concentration and sample preparation improved outcomes, and the experiments provided important insight into optimization. Overall, these experiments enhanced my technical skills, validated the protocols for further refinement, and taught me the value of careful preparation, troubleshooting, and persistence in research.
The skills, mindset, and exposure I gained will influence my future research significantly. I sincerely hope there will be an opportunity to collaborate with NIBB or return as a visiting researcher in the future. I would like to express my deepest gratitude to Prof. Takashi Ueda and Dr. Takehiko Kanazawa, who gave this opportunity and for experimental guidance, respectively. At the same time, thank you so much to all lab staff and colleagues who warmly welcomed me and supported my work.
Figure. Practicing of M. polymorpha transfer prior to further experiment.
University Würzburg, Germany
My name is Anh, and I am currently a second-year master’s student majoring in Biosciences at the University Würzburg in Germany. I had the privilege to join the NIBB Academic Research Experience Program 2025 where I spent two months as an intern in the laboratory of Professor Junichi Nakayama. It was an honor to be a part of this program, which not only expand my scope of scientific knowledge but also gave me an opportunity to experience with Japanese culture and connect with the people.
Nakayama Sensei’s lab focuses on the study of epigenetic phenomena, particularly how chromatin modifications can affect gene expression. Methylation, especially histone methylation, is one of the key modifications that can alter the “on” and “off” states of the genes. During my stay in Okazaki, I joined a collaborative research project investigating the histone modification function and the target(s) of a novel jmjC-domain containing protein in plants. Firstly, I amplified the corresponding cDNA by PCR and introduced it into a plasmid for a baculovirus expression system. The recombinant protein was expressed in insect cells to test for potential histone demethylation activity. The western blotting results showed that the novel jmjC-domain containing protein may play a role in histone demethylation against specific histone methylation, though further experiments are needed for a final conclusion. I also performed an identical parallel experiment for IBM1, another jmjC-domain containing protein involved in histone H3K9 demethylation in plant, but due to time constraints, I was not able to purify and test it.
My previous research skills mainly centered around genomic molecular techniques such as PCR, CRISPR-Cas9 gene editing, and sequencing, so this opportunity is a valuable chance for me to deepen my understanding of protein expression and acquired advanced skills like Western Blotting and affinity tag purification.
Beyond the lab work, integrating into Japanese culture and tradition was a memorable experience during my stay. Visiting Okazaki and other cities, joining local festivals and culture, and enjoying Japan’s cuisine were absolutely unforgettable.
I want to express my gratefulness to Prof. Nakayama and all the lab members. Nakayma Sensei was extremely nice, and patient and I am deeply grateful for the opportunity. I also want to sincerely acknowledge Nishimoto and Yoshimura for guiding and instructing me with the experiments, as well as Dr. Hayashi, who is so considerate and patiently troubleshoots the problems with me. All the other members were super friendly and kind. This internship has been an enriching experience, and look forward to bringing the knowledge I have gained in my professional scientific career.
Mediterranean Agronomic Institute of Chania, Greece
I’m Azoui Mouad, a Plant Genetics and Biotechnology Master’s student at the Mediterranean Agronomic Institute of Chania, Greece. I had the opportunity to be part of the NIBB Academic Research Experience Program, joining the Division of Plant Environmental Response as a research intern, under the supervision of Professor Morita. During my 1 month stay, I got the chance to work on different aspects of the gravitropic signaling in plants, using Arabidopsis thaliana, microscopy, genotyping and other laboratory techniques.
LZY4, an important protein in the gravitropic signaling cascade, was visualized using a confocal microscope. Three transgenic lines were used to study its subcellular localization, translocation, and the role of its domains. The non-modified LZY4 fused to mScarlet localized to the amyloplasts in the columella cells of A. thaliana lateral roots, where the amyloplasts functioned normally and sedimented under gravistimulation. The LZY4 fluorescent signal was also detected at the plasma membrane as a ring-shaped pattern following the initial gravity vector, which gradually disappeared and reappeared at the lower side of the root, in the direction of the new gravity vector.
The Δ1 LZY4 protein, lacking domain 1, was observed at the plasma membrane and in the cytosol of the columella cells, but no signal was detected at the amyloplasts. In contrast, the B6Q mutant, with six amino acid substitutions at the B site, showed fluorescence only at the amyloplasts and not at the plasma membrane, with the amyloplasts appearing less dynamic than in the non-modified LZY4 line. Subsequently, we can say that LZY4 translocates from the amyloplasts to the plasma membrane in response to gravistimulation, and domain I and site B are crucial for its correct subcellular localization and functional activity.
Figure 1: a: LZY4-mScarlet/lzy4, Ring-shaped signal at the start (A) and end (B) of a 15min timelapse.
b: LZY4-Δ1-mScar /lzy2;3;4 Subcellular localization. c: LZY4-B6Q-mScar /lzy2;3;4 Subcellular localization.
The rld1;2;3 triple mutant, of the RLD protein family that functions downstream of LZY proteins in the gravitropic signaling pathway, was analyzed through genotyping and phenotyping to examine segregation and Mendelian inheritance. Progeny from the rld1;rld2(+/-);rld3 cross were used for PCR genotyping (Figure 2). A Chi² test confirmed that the segregation followed a 1:3 Mendelian ratio, consistent with a single recessive locus (RLD2).
Phenotypic analysis revealed clear differences between the homozygous triple mutant rld1;rld2(-/-);rld3 and the heterozygous or non-mutated rld2 lines (rld1;rld2(+/-);rld3 and rld1;rld2(+/+);rld3), particularly in root length (Figure 3) and overall seedling development (Figure 4). The triple homozygous mutant had a severe reduction in root length and seedling development overall. This is due to the involvement of RLD proteins in the auxin transport and organ development, through the PIN auxin efflux proteins.
Figure 2: PCR results for 32 samples of the rld1;rld2(+/-);rld3 progeny.
L: DNA ladder.
Figure 3 (Left): Root length by rld1rld2(+/-)rld3 genotype.
Figure 4 (Right): Phenotypes of the 3 different progeny genotypes.
RLD1p:RLD3-ΔPH-GFP/rld1-4 T2 T1 and T2 seed selection was also conducted using hygromycin MS mediums, and the selection was evaluated using fluorescent microscopy to confirm the insertion.
To further investigate the role of LZY proteins and amyloplasts in gravitropism, the lzy2;3;4 and pgm mutants were analyzed (Figure 5). Wild-type roots grew downward along the gravity vector, showing normal gravitropism. The pgm mutant displayed a similar downward growth but with slightly more variation (Figure 7), reflecting weakened gravity perception due to the lack of starch-filled amyloplasts. In contrast, the lzy2;3;4 mutant exhibited a different phenotype, with roots growing upward and showing an anti-gravitropic response (Figure 6) caused by the disruption of LZY-mediated gravity signaling. When the pgm mutation was introduced into the lzy2;3;4 background, the anti- gravitropic response was alleviated (Figure 7). This indicates that sedimentation of starch-filled amyloplasts is essential for both normal gravitropic and anti-gravitropic responses in roots.
Figure 5 (Top Left): A. thaliana seedling of wild type (A), pgm (B), lzy2;3;4 (C) and lzy2;3;4 pgm (D) mutants
Figure 6 (Top Right): Normalized root trajectories and growth angles in A. thaliana lzy2;3;4 mutants
Figure 7 (Bottom): Radial histogram of the distribution of root growth angle in A. thaliana
To examine the Gravitropic Persistent Response (GPS), a phenomenon observed at room temperatures where the floral stem stores the gravistimulation signal at low temperatures and delays the response, Arabidopsis thaliana Col wild type and the arl2 mutant were used. The wild type displayed a clear GPS response, bending in accordance with the initial gravistimulation vector after being returned to room temperature (Figure 8). In contrast, the arl2 mutant showed no bending response under the same conditions. This aligns with previous findings identifying ARL2 as the GPS4 mutant, which lacks the ability to exhibit a GPS response.
Figure 8: Composite image of the Gravitropic persistent signal response in Wild type A. thaliana
g: Initial gravity vector
This internship gave me valuable hands-on experience in a state-of-the-art research facility and, most importantly, allowed me to further explore my main scientific interest, plant development and gravitropism. I am grateful to Morita-Sensei for this opportunity and to all the lab members, from assistant professors and technicians to graduate students, for welcoming me into their lab, teaching me patiently, and helping me throughout my stay in Japan.
Beyond the academic experience, this internship was also a wonderful chance to visit Japan, to discover the beautiful culture and history of Okazaki, and to get to know people from a different background and culture. It was an incredible experience in every aspect and left a lasting impression on me regarding Japan and its hospitable and considerate people. I sincerely thank all the lab members, the NIBB administration, and the AREP staff for making this opportunity possible, and I wish the scientists at NIBB great success in their research. I also hope I can visit again in the future.
Can Tho University, Vietnam
I recently graduated with a Degree in Veterinarian from Can Tho University, the largest university in the Mekong Delta region of Vietnam. During my studies, I developed a strong interest in symbiosis. This year, I had the privilege to participate in the NIBB AREP 2025. Joining Shigenobu sensei’s Laboratory of Evolutionary Genomics was a valuable opportunity to deepen my understanding of symbiotic relationships, particularly the ancient, obligatory symbiosis between the pea aphid Acyrthosiphon pisum and its endosymbionts.
In Shigenobu sensei's lab, my project focused on testing various protocols of mass spectrometry imaging (MSI) to investigate the spatial distribution of amino acids in biological samples. The main goal was to determine the amino acid distribution within the pea aphid tissues to gain a deeper insight into the nutritional mutualism between the aphid host and its obligate endosymbiont, Buchnera aphidicola. This project introduced me to several new areas, such as culturing pea aphids, aphid tissue sectioning and working closely with the state-of-the-art MSI. Fortunately, thanks to the dedicated and frequent guidance from Dr. Dolma Michellod and thoughtful feedback from Shigenobu sensei, my work proceeded smoothly and effectively.
My initial step was to establish the amino acid profiles in aphids to gain a general understanding of the abundance of each amino acid within their bodies. I achieved this by extracting metabolites from young aphids aged 7-10 days and subsequently utilizing Liquid Chromatography-Mass Spectrometry (LC-MS) to analyze the composition of amino acids in the samples.
The other parts of the project involved the utilization of MSI to detect and localize amino acids within aphid tissue sections. The first phase involved testing the capability of amino acid detection following derivatization with 2,4,6-triphenylpyrylium tetrafluoroborate (TPP), utilizing the conventional matrices 2,5-Dihydroxybenzoic acid (DHB) and 1,5-Diaminonaphthalene (DAN). Furthermore, various techniques for applying TPP solution onto the amino acid samples, including manual spraying with an airbrush gun and incubation within a dark chamber, were evaluated. The results indicated that DAN was the more suitable matrix for amino acid detection. Additionally, although TPP boosted the signal intensity of the amino acids, it concurrently generated significant background noise, which affected the quality of the measurement. The final step involved applying the most effective protocol utilizing DAN without the derivatization reagent TPP to tissue sections of young pea aphids. The observation of amino acid signals on the aphid tissue sections was a truly satisfying and rewarding result to me. While further optimization is required to establish a reliable MSI protocol for the localization of amino acids in aphid bacteriocytes, these achievements serve as a crucial preliminary step for a novel approach to investigate the nutritional exchange in symbiotic models.
Figure 1. Asparagine distribution in a 9-day-old aphid section
Beyond my lab work, I had an incredible opportunity to immerse myself in Japan's rich culture. My time was spent exploring Okazaki's various historical and cultural sites, savouring the local cuisine, and even attending an exciting volleyball match in the renowned SV League. Despite the cool autumn breeze, the warmth and welcome I received from the friendly lab members and NIBB staff made me feel incredibly supported.
Throughout this research experience program, my research capabilities were profoundly refined, extending beyond theoretical understanding. I was allowed to actively engage in all phases of the research project. This has significantly honed my research skills in experiment planning, time management, data analysis, logical and critical thinking. I was also inspired by the passionate and serious attitude of other lab members towards their research. The NIBB AREP was truly a wonderful experience to reinforce my passion for science, in particular symbiosis. I can’t wait to apply the skills and knowledge I gained during this program to my future research.
Hanoi University of Science, Vietnam
My name is Linh, an undergraduate student from Hanoi University of Science, and I am currently working at the Vietnam Academy of Science and Technology. I had a wonderful opportunity to join the NIBB Internship Program in 2024 and spent my internship in the lab of Professor Morita. The one-month internship provided me with invaluable experiences—not only did I acquire new knowledge and lab skills, but I also learned a great deal about work organization, data analysis, and scientific methodology. Moreover, it was truly an eye-opening experience. The working style, mindset, and passionate attitude of everyone in the lab showed me what real scientific research is like. It made me realize that I still have a great passion for research and aspire to pursue it in the future, especially in an international research environment. Moreover, what impressed me the most was the warm welcome from all the lab members. They were always willing to help me, not only with my work but also in my daily life throughout my time in Japan.
This was truly a wonderful research opportunity where I had the chance to explore and gain valuable insights into gravitropism, an intriguing response mechanism in plants that I had previously only learned about in school. For the first time, I delved into the molecular mechanisms, potential protein families, and their possible interactions that drive plant responses to gravity. Under the guidance of Prof. Morita, Dr. Nishimura, Dr. Kawamoto, Dr. Shikata, and Dr. Mori, I acquired many new skills and knowledge. From this experience, I developed a comprehensive understanding of the basic step-by-step procedures and learned how to conduct research on gravitropism using Arabidopsis as the model organism. I participated in four main research objectives. First, I learned how to evaluate the differences in phenotypes of some mutant lines and compare them to the wild-type (Col) to assess the impact of these mutations on plant morphology and predict the roles of the corresponding genes. Through this, I became familiar with key steps such as medium preparation, sowing seeds, growing plants in soil, and phenotype evaluation methods such as starch staining and measuring root tip angles. The three mutant lines I analyzed were pgm (a mutation in the gene encoding PGM-Phosphoglucomutase), lzy234 (a LAZY1-LIKE (LZY) triple mutant), and lzy234pgm. The evaluation results showed that the lzy234 mutation severely impairs the gravitropic response in roots (causing most roots to grow upward instead of following the gravitational direction), but it does not affect starch synthesis or sedimentation (as indicated by the iodine staining results at the root tips of these mutants). This suggests that the LZY protein could play a vital role in auxin polarization or other downstream signaling processes. Although PGM is involved in the interconversion of glucose-1-phosphate (G1P) and glucose-6-phosphate (G6P), a crucial step in starch synthesis, the gravitropic deficiency of pgm and lzy234pgm was less severe than that caused by lzy234. This finding suggests that mechanisms beyond the starch-statolith hypothesis might contribute to gravity sensing and root orientation, and that pgm could interfere with the impact of lzy234. Second, I learned how to select the desired transgenic lines (homozygous in transgenic genes) by analyzing the segregation ratio, which truly impressed me because it was the first time I had the chance to apply Mendel’s Law in such a clear and practical way. Additionally, I learned how to select homozygous mutants in RCC1-like domain (RLD) proteins—specifically the rld triple mutant—using genotyping. The main objective was to obtain rld123 expressing PIN2-GFP and ARF1-GFP to observe the localization of these two proteins in the triple mutants. Lastly, I performed the Yeast Two-Hybrid System for the first time to detect the interaction between the LZY1 protein and the RLD1 protein, particularly between the CCL domain (LZY) and the BRX domain (RLD). The process of conducting the experiment and evaluating potential sources of error allowed me to develop deeper problem-analysis skills and draw valuable lessons for myself. Moreover, I had the opportunity to learn how to use various types of microscopes, ranging from basic to highly advanced ones—some of which I had never seen before. All of them were fascinating and incredibly exciting. Using these microscopes, I was able to observe asymmetric root growth following the gravity direction, as well as the relocalization of LZY4-mScarlet when altering the gravity direction.
Fig1. The root growth direction of mutants is upward (opposite to the direction of gravity) compared to the wild type (Col).
Fig2. The root tip angles of wild type (Col) and mutant lines
Fig3. Starch staining of wild type (Col) and mutants
Fig4. The relocolization of LZY4-mscarlett when changing gravity direction. (A shows the distribution of LZY4-mscarlett mainly concentrated on the upper side of the cell. After changing the gravity direction (as indicated by the blue arrow both the statoliths and LZY4-mScarlet relocated to the bottom side of the cell (B))
In conclusion, this has truly been the most incredible experience I have ever had. Beyond gaining new knowledge and skills, I also learned so much about how scientific research is conducted. The lab presentations provided me with deeper insights into gravitropism as well as the work of other lab members, giving me a broader perspective on the responsibilities of PhD students in the lab. Everyone in the lab was incredibly friendly and patient with me, always willing to teach me step by step. They even introduced me to Japanese specialties and shared fascinating aspects of Japanese culture with me. Additionally, I had so many exciting experiences and trips in Japan—ones I never thought I would have the chance to see and try. I am truly grateful to Professor Morita, all the lab members, and the NIBB Committee for giving me this amazing opportunity. This is an experience I will never forget.
Heidelberg University, Germany
My name is Tiago Cordeiro da Trindade, and I am currently a first-year master student majoring in Developmental and Stem Cell Biology at Heidelberg University. I was honored to spend two months as an intern at the National Institute for Basic Biology (NIBB) in Okazaki, Japan, working under the guidance of Professor Kamei and Nonaka and their dedicated teams. This internship was a transformative experience that deepened my understanding of developmental biology and molecular regulation.
During my time at NIBB, I engaged in several projects that allowed me to explore techniques in gene editing and embryonic development. One of the primary projects involved inducing somite-specific gene editing in medaka embryos using the Gaudí toolkit and the IR-LEGO system. The objective was to achieve targeted heat shocks to induce Cre-LoxP recombination, enabling visualization of gene expression changes in specific somites.
I began by creating an F1 line of medaka embryos from a Cre driver line under a heat-shock promoter and a reporter line containing fluorescent proteins flanked by LoxP sites. This setup allowed for the observation of Cre-mediated recombination through the switching of fluorescent markers upon activation. Using the IR-LEGO system, I performed localized heat shocks on specific somites, which successfully resulted in targeted fluorescent signals (Fig. 1). Observing the precise expression of fluorescent markers in the somite cells was incredibly rewarding and affirmed the effectiveness of the techniques employed.
Figure 1. A: Negative control, no heat shock. B: Positive control, whole-body heat shock showing widespread fluorescent signal. C: Localized somite heat shock showing targeted fluorescent signal in somites 2 and 4 (Scale Bars: 500µm).
Another significant project was replicating the flow chamber experiment by Nonaka et al. to investigate left-right asymmetry in mouse embryos. The goal was to apply artificial rightward flow to pre-somite stage mouse embryos to see if it would reverse the usual left-right patterning. We prepared wild-type ICR mouse embryos and placed them in a flow chamber with the correct node orientation. Despite the meticulous setup and execution, the embryos developed with normal situs solitus. This outcome suggested that left-right determination might have occurred before the experiment, providing insights for future research.
Figure 2. Embryos from rotation culture(A) and artificial flow(B), showing situs solitus in both.
Additionally, I worked on transfecting NIH3T3 cells with the B-gTemp thermosensor to evaluate the effects of different promoters on protein expression and cell behavior.
Figure 3. A: Negative control (no plasmid). B: Positive control (CMV promoter), showing strong expression. C–E: Expression of bgTemp in Clones 1, 2, and 3, with Clone 1 showing the highest expression (Scale Bars: 50µm).
Throughout these projects, I gained hands-on experience with advanced laboratory techniques such as microinjections, flow chamber setups, transfection methods, and fluorescence microscopy. I also developed critical thinking and problem-solving skills, especially when troubleshooting experimental challenges. Collaborating with the talented researchers and fellow interns at NIBB fostered a supportive learning environment that enhanced my scientific knowledge and interpersonal skills.
Beyond the laboratory, immersing myself in Japanese culture was an enriching aspect of my internship. Exploring Okazaki, participating in local traditions, and building friendships with lab members made my stay memorable. The hospitality and kindness I encountered left a lasting impression on me.
I am profoundly grateful to Professor Kamei, Nonaka, Yoke, Hayashi, Okada and all the members of the laboratory for their mentorship and encouragement. Their expertise and patience were invaluable to my growth as an aspiring scientist. I also want to extend my sincere appreciation to the NIBB Internship Program for providing this opportunity.
The knowledge and experiences I gained at NIBB have inspired me to pursue further studies and contribute to the scientific community. I look forward to building upon this foundation in my future academic and professional endeavors.
Guru Gobind Singh Indraprastha University, New Delhi, India
My name is Tanya, and I am a postgraduate from Guru Gobind Singh Indraprastha University, New Delhi, India. Through the NIBB internship program, I had the privilege of joining Nakayama Sensei’s lab as an intern from September to December 2024. This experience was transformative, offering invaluable insights into advanced molecular biology techniques and experimental approaches.
During my internship, I focused on purifying subunit 2 of RNA Polymerase II, gaining hands-on expertise in techniques such as Tandem Affinity Purification and Western Blotting. These tasks honed my skills in sample preparation, protocol execution, and data analysis. The opportunity to work closely on protein purification and expression studies not only enhanced my technical proficiency but also deepened my understanding of the intricate processes involved in gene cloning.
Beyond the lab, the internship allowed me to immerse myself in Japan’s rich cultural tapestry. I had the chance to explore traditional customs, savor diverse cuisines, and visit various regions, all while experiencing the warmth and hospitality that made me feel like part of the community. This cultural exposure beautifully complemented my academic pursuits, enriching my overall experience.
The supportive and collaborative environment in Nakayama Sensei’s lab played a crucial role in making this journey memorable and impactful. I am profoundly grateful to Nakayama Sensei and the entire lab team for their guidance and encouragement, as well as to the NIBB program for facilitating this incredible opportunity. This internship not only broadened my academic and cultural horizons but also reshaped my approach to research and strengthened my passion for scientific discovery.
Fulbright University Vietnam Vietnam
Currently, I am a fourth-year undergraduate student majoring in Biological and Health Sciences, with a minor in Linguistic Anthropology at a small liberal arts college in Vietnam. In summer 2023, I had a great honor to join Prof. AOKI’s lab as an internship student for the NIBB internship program 2023. During my 10-week stay, I had one of the best times for learning, living and truly doing science, for which I was able to grow not just as an aspiring scientist, but also as a person. Everyone in AOKI lab never stopped amazed me with their warm and kind hospitality, their great passion and excellence in doing research. This experience had taught me some invaluable lessons, and fostered some meaningful connections that I would like to cherish!
What I learn and appreciate the most is the true scientific spirit that everyone in the lab cultivates. Under the guidance of Dr. SUGIYAMA, Dr. GOTO, and Prof. AOKI, I was trained to ask critical questions for my research, with a healthy and respectful skepticism. I have learnt to pay great attention and care to my works. They taught me from the smallest details, such as efficient pipetting skills or labeling my samples, to the most general concepts about science, academia, and life. I learnt the most basic to the cutting-edged techniques in biology, from simple gel electrophoresis or Western blot to complicated cell imaging processes. This internship has shown me great reflections about my research career. I think I have made the right choice, and now I am more than ready to pursue a scientific career ahead.
One of the main projects in which I was involved was to study the effects of nutrient conditions on division and morphology of S.pombe. To do that, I conducted time-lapse experiments of fission yeasts, whose CDK activity (i.e., phosphatase and dephosphatase activity) was measured by a newly developed FRET, in 02 nutrient conditions (i.e., rich vs poor in Carbon and Nitrogen source). I experimented on 2 yeast strains that were previously designed by Dr. SUGIYAMA. Fission yeasts in each of these strains were tagged with other phase-marker proteins (i.e., PCNA, Pom1, and Cdc13). This system allowed me to quantify the temporal dynamics between CDK activity and other related proteins involved in the governance of cell division cycle.
The results we found provided preliminary evidence that nutrient conditions did modulate changes in the cell cycle progression, reflected via (1) cell cycle duration, (2) CDK peak activity, (3) S-phase duration (Figure 1: Top, Middle, Bottom, respectively). In that, although the overall cell cycle duration of fission yeast in Poor condition was longer than that of Rich condition, the duration of S-phase was significantly extended in the latter condition. It was possible that S-phase in Poor condition was shorter due to the lack of nutrients, so that G1-phase, possibly G2-phase, was elongated as a compensating mechanism in Poor condition. In addition, we also found some complex changes in the morphology of Pom1-deleted cells, in response to the nutrient conditions. The clear relationships and mechanisms await further in-depth studies and analyses.
Figure 1. Cell cycle progression (Cell cycle duration – top; CDK peak – middle; S-phase and G1-phase timing - bottom), compared between Poor and Rich condition.
To see the cells progress through their cycle, with different protein expressions tagged with many fluorescent proteins, and to be able to analyze the results are truly fascinating to me. Additionally, I also learnt to clone DNA and design plasmid to create a new S.pombe strain whose heterochromatin assembly can be optogenetically manipulated. I was also able to participate in one small optogenetic experiment where the ZO-1 protein of HeLa cells was turned on and off by different lights. I literally wow-ed when the protein translocated to the cell’s membrane immediately after the light was shined onto the HeLa cells. The response was just in a blink of an eye. It was just WOW!
Once again, I am truly grateful for the opportunity. I would like to express my sincere thanks to everyone in AOKI lab. My time there was great, and I am missing you all already!
Kyushu University, Japan
My name is Trang Thu Dao and I’m a final-year student at the Department of Bioresource and Bioenvironment, Faculty of Agriculture, Kyushu University, Japan. I am very grateful to have the opportunity to do a two-month internship in the Division of Chromatin Regulation (Professor Jun-ichi Nakayama’s laboratory). As an NIBB intern, I joined part of Dr. Aki Hayashi’s project on histone modification. It was an immensely enriching experience that allowed me to delve into the fascinating realm of epigenetic inheritance and CRISPR-Cas9 gene editing.
In the scope of epigenetic changes, environmental stress leads to modifications to the DNA molecule or associated proteins that can be passed on to subsequent generations without altering the underlying DNA sequence. One of the major epigenetic information carriers is heterochromatin. In the case of heterochromatin components, the methylation status of H3K9 is crucial in epigenetics, regulating gene expression to cope with environmental stress. If a gene is turned off through histone modifications in a parent cell, its offspring cells might inherit the same gene-silencing pattern. Therefore, the big question is “How do histone modifications cause epigenetic inheritance for the next generation?”
Analysis of a mechanism of transgenerational epigenetic inheritance is the key to answering this question. During my time at the Division of Chromatin Regulation, I joined a project to integrate a tethering system into the genome of fission yeast Schizosaccharomyces pombe (S. pombe) by CRISPR/Cas9 system. The tethering system regulating genomic activity will be expressed during meiosis to study the mechanism of transgenerational epigenetic inheritance.
I constructed the pCas9-gRNA plasmid to create a DNA double-strand break for insertion by homologous recombination. After preparing donor DNA fragments for integration, I performed genome editing on S. pombe to insert the gene encoding the tethering system. I also analyzed the localization of telomeres in fission yeast spores under the microscope. As Taz1 is a subunit of the telomeric complex, it is used to fuse the GFP protein for localization. The telomeres were observed inside the fission yeast spores.
Localization of telomeres in fission yeast spores using the Fluorescence Microscope
Along with learning new experimental techniques, I had opportunities to attend Joint Progress seminars, in which I could learn more new concepts and insights about histone modifications. The final presentation equipped me with the experience of professional scientific communication. I am very grateful for all the invaluable experiences in NIBB.
After all, I would like to express my appreciation to Prof. Nakayama, Dr. Hayashi, and all the members of the Division of Chromatin Regulation for their warm welcome and wholehearted support to me during my internship. I would like to extend my special thanks to Dr. Kiyoshi Tatematsu for his support to prepare administration documents. Last but not least, I also want to express my gratitude to the NIBB Internship Program for giving me this chance to experience research and life in Okazaki City.
Hanoi University of Science, Vietnam National University, Vietnam
This three-month internship at Prof. Uchiyama's Lab has been a pivotal experience, marking a significant upgrade in my research skills. As an experimental biology student, I had not been familiar with programming. Working in this lab was the first time I had learnt about the UNIX environment. I also had a chance to practice my coding skills with R and Python language and how to apply them in genome analyzing, a skill set I hadn't explored before. My entry into Uchiyama Labs was driven by the challenges encountered during my bachelor's thesis, particularly my inquiries into genes associated with sulfur oxidation pathways. Prof. Uchiyama's warm reception and enthusiastic support were instrumental in shaping the direction of my work. Using various bioinformatics approaches, I explored the co-occurrence and other strong relationships between FCC and SOX systems (Fig. 1–3), which are crucial genes involved in sulfur oxidation. Another significant result is my insights into the characterized functions of distinct subgroups within the sulfide-quinone reductase gene, through MBGD data, HMM classification, phylogenetic tree, phylogenetic profile and neighborhood genes analysis. Despite my novice status in the "dry lab," Prof. Uchiyama's persistent guidance and kindness helped me become more confident. Besides that, I am very thankful to all members of the Genome Informatics Laboratory and The Data Integration and Analysis Facility. They were incredibly enthusiastic in supporting me to resolve my research issues without any hesitation. Also, recognizing my interest in gacha items, they kindly presented me with numerous ones as gifts before my departure to Vietnam.
Figure 1. Correlation coefficients among phylogenetic profiles (a) and map comparison (b) show the relationship between FCC and Sox system
Figure 1. Correlation coefficients among phylogenetic profiles (a) and map comparison (b) show the relationship between FCC and Sox system
Gyeongsang National University, Korea
My name is Yeongcheon Yang. I graduated from Division of Life Science of Gyeongsang National University in Korea. Like many students, I was also worried whether my career path (direction of research) would be the same or different as I thought. However, the NIBB internship program not only was an opportunity to relieve this anxiety, but also an incredible experience that was priceless. Through this program, I got a chance to take part in Professor Morita lab and study their current experiment as a research student. I was able to learn about the gravitropism and how this mechanism works in plants.
It is true that my research in NIBB internship program mainly about the gravitropism, but I was possible to learn fundamental biological experiments from the selection process of mutants, which are the basis for research, to the interpretation of data. During the 3 weeks internship, I focused on three major things. First, selection of mutant plants. Second, relationship of LZY and Amyloplast. LZY is a key in gravity response of plant, and LZY stick to amyloplast’s outside surface and during the gravity stimulus move to plasma membrane from there. Lastly, the function of Amyloplast and LZY. Each LZY2, LZY3, LZY4 and PGM function was inferred from the phenotypes of single and double, triple, and quadruple variants.
To select specific mutant plants for research, I used antibiotic resistance selection (Kanamycin, Hygromycin B) and fluorescence methods. In this process, I was very surprised to find that Mendel's genetic laws, which were only seen in books, were very useful.
To identify phenomena occurring in columella cells during gravitational reactions, LZY4 protein containing mScarlet fluorescent protein was constructed (LZY4p:LZY4-mScarlet/lzy4). Plus, in order to compare the polar movements of LZY between normal and abnormal amyloplast, (LZY4p:LZY4-mScarlet/lzy4·pgm) was constructed. pgm, a key enzyme for starch synthesis, is essential for Amyloplast sedimentation and redistribution. Consequently, the pgm variant showed a large difference compared to plants with successful amyloplast precipitation and distribution of LZY4.
Figure 1. Left LZY4p:LZY4-mScarlet/lzy4. Right LZY4p:LZY4-mScarlet/lzy4·pgm. LZY4 colored magenta. In both plants, the direction of gravity is downward. Interestingly, Amyloplast in columella cells LZY4p:LZY4-mScarlet/lzy4·pgm does not show proper sedimentation. Moreover, LZY4 are not present in polarity.
Figure 2. The different root phenotype of Col-0 and lzy2·3·4. Unlike Col-0, the root of lzy2·3·4 extending upward as a whole.
Through the function of LZY and the indirect loss of amyloplast sedimentation of pgm mutants, I could guess the function of LZY affects to downward root growth. Furthermore, I could understand LZY plays a pivotal role in gravitropism more than I thought.
Figure 3. Starch stained with 4% Idoine at Col-0 and lzy2·3·4·pgm. In contrast to Col-0, lzy2·3·4·pgm is not stained because Amyloplast does not have starch.
Figure 4. The main root angle difference of each variant with Col-0. Col-0 and pgm variant shows downward root extension. Conversely, lzy2·3·4 roots toward upside. Interestingly, lzy2·3·4·pgm show most variety main root angles.
Overall, NIBB internship program not just helped me to learn a new concept, also to understand how valuable basic knowledge and methods is. I appreciate to all members of Prof. Dr. Morita laboratory, other technicians, and responsible people of this program. I am very grateful to them for providing to me precious opportunity to be in this program in Japan. Of course, I hope to back to Japan in the future, as well.
Hanoi University of Science and Technology, Vietnam
Hello everyone! I’m Nguyen Yen Chi, a recent graduate from Hanoi University of Science and Technology (HUST). I had the wonderful opportunity to participate in a 3-month internship at the Evolutionary Genomics Laboratory, under the guidance of Professor Shuji Shigenobu. Before this program, my expertise mainly centered around wet lab. Knowing that Shigenobu Lab provides training on both experimental and dry lab skills, I was immediately intrigued to expand my knowledge in bioinformatics and genomics techniques. This invaluable experience was an essential period that shapes my future.
My project centered on Regiella insecticola, a facultative endosymbiont of aphids. Among the three common secondary symbionts in pea aphids – Hamiltonella defensa, Serratia symbiotica, and Regiella insecticola – the last one has been less extensively studied. Hence, my objective was to broaden the understanding of R. insecticola, particularly concerning its localization, transmission, and interactions with both the primary symbiont, Buchnera aphidicola, and the host insect. This was the first time I have ever worked with insects, offering me numerous novel experiences.
Initially, I detected the presence of this bacterium in several pea aphid strains in our lab through diagnostic PCR. Using fluorescent staining and FISH techniques, I observed numerous cells and organs in both viviparous and oviparous female individuals. The resulting images helped me localize the secondary symbiont and uncovered several findings regarding the transmission of R. insecticola throughout sexual and asexual generations. There were both novel observations and results that aligned with previous descriptions about other secondary symbionts. In the strain of pea aphid I studied, Regiella resides not only within the sheath cells but also outside of these cells and the bacteriocytes. Additionally, they were observed surrounding the fat cells and present inside some fat cells.
Fig 1. Bacteriome structure illustrated by Nozaki & Shigenobu (left) and FISH image showing a part of bacteriome of pea aphid strain 8 (right). Buchnera is red; Regiella is yellow-green.
In my exploration of Regiella's characteristics and interactions, I familiarized myself with numerous command-line tools such as seqkit and local BLAST, conducted KEGG analysis, and utilized IGV for genome visualization. My skill set was expanded beyond the tools learnt in my bachelor's program. Additionally, I had the chance to take a Bioinformatics course in November, which provided an enlightening overview of the field through hands-on experiences. For me, this subject was new and challenging, and I received a lot of help from my lab members. As a result of new bioinformatics knowledge, I could discover new characteristics regarding the affection of Regiella to pea aphid and Buchnera. For example, it appears that Regiella does not fully compensate for Buchnera in many amino acid biosyntheses. In the specific strain that I studied, Regiella seemed to have positive affection on the growth speed of host aphids.
Throughout my internship at Shigenobu Lab, I was fortunate to receive invaluable guidance and support from all lab members. I have admired and learned a great deal from their work ethics, their knowledge, and their broad perspectives. I owe a debt of gratitude to Dr. Tomonari Nozaki, our lab's assistant professor, for his guidance in my lab work as well as his assistance in my social life. Beyond a mentor, he and his family are precious friends of mine. Overall, I genuinely feel blessed to have crossed paths with such exceptional people.
For all these priceless experiences, I am incredibly grateful to Professor Shuji Shigenobu and NIBB Internship Committee for accepting me to the Internship Program 2023. This experience allowed me to work and learn under Professor Shigenobu's guidance in his laboratory, granting me a broad range of knowledge and practical skills in an entirely new field of study. Furthermore, I've had the honor of meeting intelligent, kind-hearted people and building lasting friendships. This time become an invaluable journey filled with learning, personal growth, and unforgettable memories that will always be dear to me.
University of Heidelberg, Germany
My name is Bal, and I am a master`s student of developmental and stem cell biology at the University of Heidelberg in Germany. Through the NIBB internship program, I got to join Kamei sensei`s lab and worked at the optics and imaging facility. This enriching experience spanned from December 2022 to February 2024 and provided me with invaluable insights into various microscopy techniques and experimental design. During my stay, I immersed myself in learning state-of-the-art microscopy techniques such as Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM), micro-CT, Selective Plane Illumination Microscopy (SPIM), etc. The hands-on experience allowed me to gain proficiency in sample preparation, imaging, and data analysis associated with each technique. In addition to microscopy techniques, I further refined my skills in tissue culture and microinjections.
My research primarily focused on utilizing Medaka and HeLa cells as models for studying different biological processes. For example, I utilized scanning electron microscopy (SEM) to visualize the micropyle in Medaka embryos. Additionally, I conducted experiments using the Gaudi (Hsp-Cre) * TG861(bactin-loxp-DsRed2-loxp-GFP) Medaka line. In response to IR LEGO, the region originally expressing DsRed2 turned green upon Cre induction, illustrating the dynamic changes in gene expression.
Beyond the laboratory, my internship experience extended to exploring the rich cultural tapestry of Japan. I had the opportunity to immerse myself in different cultural traditions, savor diverse cuisines, and explore other regions. The warmth and hospitality I experienced made me feel like part of the family. The supportive lab environment and cultural exposure facilitated a well-rounded learning experience.
In conclusion, my internship at Kamei Sensei's lab within the Optics and Imaging Facility was an enriching journey that reshaped my approach to science and broadened my horizons both academically and personally. I am immensely grateful for the opportunity and the invaluable lessons learned, which will undoubtedly guide my future research endeavors. I am thankful to Kamei Sensei and the entire lab team for their guidance and support throughout my internship and to the NIBB program for facilitating the internship.
University of Chinese Academy of Sciences, China
I am Kang Lyu. As a master graduate from the University of Chinese Academy of Sciences, I am passionate and interested in NIBB's internship program. Through several weeks’ internship in the institute, I further felt the charm of scientific exploration at Professor Jun Minagawa’s laboratory of Division of Environmental Photobiology. It is my honor to have both Professor Minagawa to be my supervisor and Dr. Kamada to be my mentor. Prof. Minagawa is an internationally renowned expert in the field of microalgae photosynthesis and enjoys a high reputation. From him, I saw his enthusiasm for research and advanced scientific thought. Dr. Kamada has made many original explorations in the field of photoprotection regulation and has achieved outstanding achievements. I saw her rigorous and realistic attitude towards scientific research. I benefited a lot from communicating with them in the field of photosynthesis research.
During my internship, I was mainly engaged in exploring the regulatory mechanism of Chlamydomonas reinhardtii photoprotection. An earlier work in the Minagawa lab showed that Algal photoprotection is regulated by the E3 ligase CUL4–DDB1DET1. The putative E3 ubiquitin ligase CUL4–DDB1DET1 in unicellular photosynthetic organisms may mediate blue-light signals to influence photoprotective gene LHCSR1 and LHCSR3 expression, thus exercising the role of photoprotection regulation. Besides, another result revealed CONSTANS flowering complex in the green alga Chlamydomonas has an important function in controlling the response of photoprotection. On the basis of the above studies, we look for other possible mechanisms that regulate the response of photoprotection.
Last but not least, I would like to express my sincere thanks to Professor Minagawa and Dr. Kamada and Dr.Makio and Dr. Kim and Ms. Yuka and other members of the laboratory. I enjoyed the time in NIBB very much. Although the time was not long, I felt the ample care and concern in such a harmonious family and felt the charm of scientific research. I would also like to thank Dr. Tatematsu and the NIBB Office for their thoughtful assistance in visa processing and during the internship. I hope I will have another opportunity to pursue scientific exploration here in the future.
University of Michigan, USA
Even though it was a time of uncertainty due to the COVID-19 Pandemic, I was warmly welcomed into the Aoki Lab for 5 weeks in July and August 2022. As a participant in the NIBB Internship Program, I worked closely with Dr. Ryosuke Tany and Dr. Yuhei Goto to help develop a new biosensor system to visualize and quantify intracellular GPCR signals via creating a new human cell line. Currently, I am a rising senior undergraduate at the University of Michigan College of Literature, Science, and Arts with a focus on Biological Sciences. As a biology major and having a previous interest in Japanese culture, I was intrigued by the prospects of living and conducting research abroad in Japan. I was able to grow not only as a researcher, but also as a person by exposing myself to a new environment and forming connections with a new culture and people.
Having worked with the Aoki Lab for five weeks, I was able to become proficient in wet lab techniques including plasmid construction, transfection with LentiX-293T, infection of HeLa cells, and live cell imaging. Thanks to the exemplary teaching by Dr. Tany and Dr. Goto, I was able to successfully create fluorescent biosensors for Ca2+, RhoA, KTR, and cAMP. In previous experiments, a biosensor system for these intracellular GPCR ligand binding activation signals was created; Ca2+ by R-GECO1 (Red Fluorescence), cAMP by EPAC-FRET (Cyan-Yellow Fluorescence), ERK by ERK-KTR (Near Infrared), and RhoA by DORA-RhoA (Cyan-Yellow Fluorescence); however, there was a problem with utilizing the same Cyan-Yellow fluorescence for cAMP and RhoA.
Together with the Aoki Lab members, we developed a new system for visualizing these intracellular signals that contains no overlapping colors; Ca2+ by B-GECO1 (Blue Fluorescence), cAMP by G-Flamp (Green Fluorescence), ERK by ERK-KTR (Near Infrared), and RhoA by dT-2xGBD (Red Fluorescence).
Fig 1. Treatment of Cocultured R-GECO and B-GECO with 10uM Histaminecells at 5 minutes for a total of 30 minutes
First, we wanted to confirm that B-GECO could be used as a Ca2+ sensor similar to R-GECO, and this was confirmed by the data analysis of co-cultured B-GECO and R-GECO cells. We found that B-GECO and R-GECO were similar as they both showed an initial peak for the influx of calcium (Fig 1.), and showed similar pulsing of cells. Next, we wanted to compare ERK-KTR-miRFP703 and ERK-KTR-iRFP, and the results were quite similar (Fig 2&3.); however, ERK-KTR-iRFP live cell imaging showed better differentiation of the nucleus and cytoplasm, so it was chosen as the better biosensor to be used in the bulk cell line.
Fig 2&3. Comparison of ERK-KTR-miRFP703 and ERK-KTR-iRFP treated with 10ng/mL EGF at 5 minutes for a total of 30 minutes
Lastly, when developing a RhoA sensor, we decided to use dT-2xGBD as our biosensor of choice, as it was able to detect cytoplasm RhoA localization to the plasma membrane.
However, this was only possible in cells transfected and expressing the GPCR BDKRB2, a bradykinin receptor. The quantification of the live cell imaging of RhoA activation was interesting as it was able to show slivers of RhoA activation in the plasma membrane.
Fig 4. dT-2xGBD expression in BDKRB2 expressing cells treated with Bradykinin at 5 minutes for a total of 30 minutes
While the research was important, I also have fond memories of discussing a variety of old and new anime and manga with the other members of the lab. Every Thursday, I would go out to dinner with a few lab members and play Badminton with other research members afterwards. Although I was never very good at Badminton, I was able to meet new people and make new friends. I also was able to play soccer and explore Okazaki City and its numerous cultural and historical landmarks, and got better at Japanese due to conversing with the Aoki Lab members who were also able to help me study and become better at my Japanese grammar and increase my vocabulary.
I loved sampling the different Japanese, Chinese, and even Korean cuisine in the city of Okazaki; one of my recommendations would be the Japanese maze-soba or Nene Fried Chicken near the Higashi Okazaki Station. During the weekends, I enjoyed taking the train or Skinkansen and exploring the vast cities of Nagoya and Tokyo, which I highly recommend. Having a farewell celebration at the end of my time in the lab, I was able to present all my research throughout the internship, and am forever grateful for the opportunity to research with the Aoki Lab and would like to thank the NIBB Internship Program and Aoki Lab.
Vietnam Research Station, Nagasaki University ‘s Project, Vietnam
My name is Pham Hong Quynh Anh. I have just finished my Master course at The University of Tokyo last April 2022. I am grateful to have an opportunity to participate in NIBB 2022 internship program. It is very great to participate in experience activity in the Lab of Genomic Informatics, under Uchiyama sensei’s supervision. My intended field of study in the host lab is to select specific tools for study on microbial comparative and evolutionary genomics. It may give me a new aspect in my current concern, how to derive gene ortholog-based sequence features for protein variants and essential mobile genetic elements and/or have sequence features common to known genomic islands for bacteria survival in different environmental conditions.
During one month in Okazaki, I did learn how to effectively access bioinformatic tools and manage my database via VPN and BIAS system; and analyze gene ortholog-based sequence features via Microbial Genome Database (MBGD) which has been developing by Professor Uchiyama’s laboratory, by using my AMR samples collected from wastewater in Vietnam.
Orthologous clustering of all Aeromonas strains identified 7971 orthologous groups (OGs) defining the pan-genome. Histogram shows 2438 OGs which are conserved in all 21 strains, and represent the universal core among them. The remaining 5533 OGs correspond to the accessory genome, in which 173 OGs are unique in only NUITM-VA1. Compared to the reference, NUITM-VA1 strain lacks 7 proteins, which related to transport & binding of protein and transcription functions (Fig1).
Fig1. MBGD
Ortholog Cluster Overview.
Two NUTM-VA1 and NUTM-VA2 strains share 9 unique genes, which are absent in all Aeromonas strains in MBGD database. Besides, these strains were collected from urban drainage which located near some center hospitals in Hanoi, this clue suggests that these unique genes have some potential function to support bacteria survival in extreme environmental conditions (Table 1).
Table 1. Unique genes found in NUTM-VAs strains
Next, I conducted an experiment to identify horizontal gene transfer (HGT) acquired genes encoding proteins that function under particular conditions, such as genomic islands (GIs).
The results found 440 blocks related to mobility classes among 50 Aeromonas strains, partly shown in Fig2.
Fig2. Overview of FindIsland output.
A black line represents a direct adjacency, a green line represents a non- adjacent neighborhood (i.e., there are insertions), and a red line represents an inversion.
The analysis conducted during this internship will be continued for the further analysis, to comprehensively answer my own question: “Which HGT and mechanism mainly affect the survival of bacteria in wastewater and resistant to antibiotics?”
In conclusion, NIBB Internship Program 2022 is an invaluable experience for me to consider entering the Ph.D. course in my next step career path. Despite the lack of time, I would like to express my sincere gratitude to Professor Ikuo Uchiyama, who accepted me to join his laboratory during my internship course. I wholeheartedly thank his patience, motivation and immense knowledge. Furthermore, I am thankful to Jiang-Kun, Kotani-san and her husband, for their kindly welcome to me to discover Okazaki sightseeing with historical views and yummy traditional foods. Special thanks to Prof. Kiyoshi Tatematsu for helping to prepare all documents. I also would like to send best regards to all staff in Uchiyama’s Laboratory and Mishima Lodge, who supported me during my one-month internship. Last but not least, many thanks are sent to my supervisors in Vietnam, my boyfriend in Tokyo and friends - alumni in NIBB, Okazaki, Aichi, Japan.
Osaka University, Japan
I am Piyusha Mongia, a graduate student from Osaka University, Japan. I am grateful that I could take part in this internship program and be apart of Prof. Jun-Ichi Nakayama’s laboratory. The lab works on epigenetics- how it regulates gene expression, the establishment of epigenetic markers and so on.
The bigger question was to understand how epigenetic information is inherited and what are the factors involved. Epigenetics refers to heritable changes- not coded by DNA. Eukaryotic chromosomes can be divided into 2 broad categories: euchromatin and heterochromatin. Euchromatin refers to the regions on chromosomes which are transcriptionally active. Heterochromatin, on the hand, is generally transcriptionally silent, present at repetitive sequences or transposons. Methylation of lysine 9 of histone H3 (H3K9me) is a hallmark of heterochromatin. I created and assessed the involvement of four candidate gene deletions in heterochromatin inheritance in fission yeast spores. Fission yeast is a simple eukaryote having three chromosomes, making its study relatively easier. I first replaced the 4 genes with nourseothricin by PCR and then visualized swi6-egfp localization by fluorescence microscopy. Swi6 is a heterochromatin protein that is required for its assembly. One of the candidate genes seems to be important for the inheritance of heterochromatin but more studies need to be done to conclude so. In a parallel project, I tried to express three of Tetrahymena proteins using a bacterial expression system. These proteins, HPL4, HPL5, and JUB5 are involved in heterochromatin assembly. Producing recombinant proteins allows us to purify and biochemically analyze individual proteins activities and interactions in-vitro. In the end, I managed to express all, but purify one of the proteins due to time constraints. I learned many biochemical techniques, like Gel Filtration Chromatography and Ion Exchange Chromatography. I wish I could have had a longer time period to do more fun experiments.
Every day I was faced with challenges, and learnt something new. All the members were very patient with me and taught me something or the other. I’m extremely grateful to Dr. Hayashi and Dr. Kataoka, who guided me throughout the project. Prof. Nakayama was very supportive and encouraging. The research atmosphere in the lab is very exciting and it was truly an enriching experience for me.
Kyushu University, Japan
My name is Geofanny Bernadette Yohanes and I am 3rd year undergraduate from Kyushu University. I came to NIBB on February 2022, after many delays due to COVID-19, and spent two weeks with Prof. Kiyoshi Naruse in the Laboratory of Bioresources.
Although I did not participate in ongoing research, I was able to do three basic experiments. I did most experiments with the laboratory staff. The first was a knock-out of SLC45a, a gene controlling melanin synthesis, into medaka (Oryzias latipes). The second, also with medaka, was a knock-in experiment of mNeonGreen to the myosin heavy chain, creating a fluorescent fusion protein. Through both of these experiments, I learned and practiced techniques such as microinjection, DNA extraction, and PCR. At the end of each day, I was given a lecture by Prof. Naruse to prepare for the next step. This allowed me to gain a deeper understanding of knowledge I have learned in school, such as the mechanism of CRISPR/Cas9, differences between gene editing methods and when to apply them, and factors to consider in primer design. It was fun to watch the embryos grow and display the results of these experiments, although some must be used for genotyping to confirm the success. The third experiment was cryopreservation of medaka sperm and artificial insemination using the preserved sperm.
Result of knock-out experiment. Melanin formation in the eyes was affected.
In addition to learning knowledge and skills specific to each experiment, this internship gave me an insight on how to be a good researcher. I learned how important it is to keep an organized journal, to clean up and take care of your tools, and what discussion around research is like.
I would like to thank Prof. Naruse and the lab staff for hosting me during this internship. It has been an invaluable experience. The skills I learned will no doubt be useful for me as I plan to work in research related to fish in the future.
Kyushu University, Japan
In my second year of undergraduate school at Kyushu University, I got accepted into NIBB internship program under the supervision of Professor Kazuhiro Aoki (Division of Quantitative Biology). This is my first time doing my own project, from designing an experiment to making a final presentation, which equips me with many useful experimental technics and introduces me to what the life of a researcher is like.
The purpose of my project is to verify the mitogen-activated protein kinase (MAPK) pathway by using two systems: optogenetics Cryptochrome-2 (CRY2) and Kinase Translocation Reporter (KTR). Upon photoexcitation (blue light), CRY2 proteins simultaneously oligomerize9 and bind to CIB1, which form clusters by promoting interconnection among CIB1-conjugated MPs (Figure 1).
Figure 1. CDK1 kinase encoded with CRY-2 formed cluster when exposed to blue light.
When being activated, KTR will move out of the nucleus, resulting in a decrease in nucleus intensity (Figure 2).
Figure 2. Nucleus intensity decreases as a result of KTR activity.
The growth factor can activate ERKKTR whose cytoplasmic relocation will make the nucleus darker. As shown in the left graph, a significant intensity drop was captured when EGF was added at 10 min.
We first selected eight kinase plasmids: RAD1, MEK, KSR1, JNK1, ASK1, TAK1, p38a, and CDK1. Then I introduced those plasmids into the CRY-2 template by LR gateway reaction. The integrated plasmids will be transfected into Hela cells that have four types of KTR inside, ERK, AKT, JNK, and p38a, respectively. To visualize kinases behavior under a microscope, eight plasmids were encoded with mScarlet, a red fluorescent protein, and four types of Hela cells with mKO, an orange fluorescent protein. If MAPKK or MAPKKK can activate MAPK, the droplet will form and there will be a dark change of nucleus; if there is no relationship between two types of kinases, only cluster formation will be captured. For example, MEK1 cluster was captured and at the same time, a change of JNKKTR nucleus had been found, which identifies that MEK1 can activate JNK (Figure 3).
Figure 3, RFP and hmKO record when exposed to blue light. Cluster formation indicates that MEK1 was activated and the change of Hela cells nucleus with JNKKTR implies JNK was activated.
Apart from the scientific project, I want to say thanks to all lab members and students in Aoki sensei’s lab. Even when back to Kyushu, I still dreamt about the day we ate Nagoya fried chicken and talked until 10:30 p.m., the restaurant we enjoyed donuts together, the whiteboard had many Hokkaido maps on it, and lunch boxes that stored our short talk every day. Many memories pop up in my mind when thinking of Okazaki. I learned a lot from their attitudes and determination towards science and become firmer to be a person like them in one day. The internship here is not one sentence on my resume, but something that can push me to move forward, and I will cherish it for the rest of my life.
Imperial College London , UK
I immersed myself in the NIBB internship program as an undergraduate student who had just completed her first year of Medical Biosciences degree at Imperial College London. Although I had limited experience in conducting experiments at a professional laboratory, I was motivated to intern at Professor Aoki`s laboratory specializing in quantitative biology upon reading one of his publications. Among the other host labs, Professor Aoki lab`s unique approach in presenting scientific results through microscopy visualizations and precisely quantified data appealed to me as an opportunity to obtain skills in both practical research and scientific communication. Throughout the course of the next month and a half, I was able to get a deep insight on using advanced techniques and equipment to visualize and model various intracellular mechanisms.
My research primarily focused on examining the proteins involved in controlling the cell cycle of Schizosaccharmyces pombe, otherwise known as fission yeast. The progression of biochemical events from G1 to M phase in its cell cycle is known to be regulated through a single CDK-cyclin complex composed of Cdc2 and Cdc13, alongside the activation of additional protein substrates at distinct timepoints.
To examine such properties, I first integrated GFP, a green fluorescence protein, at the 3’ end of a chromosomal open reading frame so that the resulting gene construct will fluoresce in varying intensities depending on the protein concentration. Confocal fluorescence imaging was subsequently conducted to examine the oscillations in fluorescence intensity over the duration of 16 hours. Moreover, I examined the expression of genes tagged with mNeonGreen, a fluorescence marker that is known to be up to five times brighter than GFP. This enabled me to visualize the expression patterns of genes such as cdc25 that fluoresce at much lower intensities.
Fig 1. Changes in the expression level of fluorescence-tagged genes throughout the progression of cell cycle were examined. mNeonGreen localized at the nucleus for cdc13, cdc2, and cdc25, while localization at the cytoplasm was observed for the reference gene, adh15.
The resulting data obtained was quantified using the ImageJ software and graphed to present the time-specific changes in protein concentrations. This allowed deductions to be made on the roles and properties of individual proteins in regulating the cell cycle. The research can be further developed by attaining additional measurements such as protein synthesis and dissociation rates which are involved in an ordinary differential equation that models the S. pombe’s cell cycle. This will enable the representation of the molecular mechanisms involved in the cell cycle of S. pombe through a mathematical model with numerically quantified parameters.
Fig 2. Time-dependent oscillations in the concentration of proteins involved in a S.pombe`s cell cycle were graphed. The red markers indicate the point of nuclear division at the end of mitosis. The Cdc2-Cdc13 MPF complex controls the progression through the different stages of a cell cycle, while Cdc25 dephosphorylates and thereby activates MPF at late G2 phase to promote entrance into the M phase. Data was obtained through live cell imaging on the Onix2 system and subsequently graphed on Python.
Fig 3. Two full cell cycles completed over the duration of approximately four hours were modelled.
The point at which the cell begins its cell cycle, shows maximum fluorescence, and undergoes nuclear division were measured and averaged to be graphed on Excel. A decrease in concentration in the following order of proteins can be deduced from the graph: Cdc2, Cdc13, Cig2, Cdc10, Cdc25, and Wih5.
Whilst being engaged in using various imaging tools, I was also introduced to basic laboratory techniques such as western blotting, transforming plasmids into cell lines, designing PCR primers, and conducting swift but precise DNA work. Additionally, I learned about the importance of performing detailed experimental planning, careful observation, interactive discussions with other researchers, reliable data analysis, and effective troubleshooting to develop as a well-established researcher capable of presenting influential scientific discoveries.
Most importantly, I want to thank all members of Professor Aoki’s lab for allowing me to be immersed in the internship program in a warm, welcoming, and ever so supportive environment. My questions were never responded with a sigh or ignorance, but instead with detailed explanations and demonstrations.
Undoubtedly, the internship program provided me with an opportunity to further explore Japanese culture by encountering people with different values, interests, and lifestyles. Interestingly, these differences were especially evident in our lunch menus. In contrast to my lunch box which was composed of Mac n cheese, oven-grilled chicken and a clubhouse sandwich, the people at the laboratory packed their “bentos” with “yakisoba” or stir-fried noodles, “karaage” or Japanese fried chicken, and “onigiri” topped with “furikake” rice seasoning. Although there were differences in our diets, the conversations held over the lunch table allowed me to broaden my perspective on cultural diversity.
Waking up in the morning to be greeted by the sounds of birds chirping, taking a refreshing 7 minute walk from the Mishima Lodge to the laboratory, occupying myself with scientific wonders alongside other researchers, and going back to the lodge at the end of the day to try out different Japanese recipes for dinner… such routine filled my days at Okazaki with joy and excitement. I want to thank the NIBB for providing me with such invaluable opportunity, and the people at Professor Aoki’s laboratory for their unconditional support throughout the internship. I am confidently returning home with newly acquired skills and attributes that have equipped me to continue pursuing my passion in research.
University of Heidelberg, Germany
Bettina Welz
My name is Bettina Welz and I am studying Molecular Biosciences with the major Developmental and Stem Cell Biology at the University of Heidelberg in Germany. Within the NIBB internship program, I had the chance to stay 2-3 months at Prof. Kiyoshi Naruse’s research group. The laboratory focuses on the evolution of the sex determination system with the Japanese rice fish medaka (Oryzias Latipes) as a model organism. Medaka has deep historical roots in Japan, if you pay attention you can find medaka in rice fields, in indoor water gardens or small ponds in front of houses just by walking down the streets. Beside its worldwide popularity among aquarists, it carries many beneficial characteristics for research, like a small genome size, high fecundity, and an ex utero development - making it a good model organism for developmental biology and genetics.
During my time at the NIBB in Okazaki I was introduced to new laboratory techniques and the handling of medaka in general. Moreover, I had the possibility to discuss and exchange opinions and experiences with leading researchers in the medaka research field. Besides performing a linkage mapping experiment, in which the relative position of genes to each other was analyzed based on recombination frequency, I performed cryopreservation of sperm and artificial insemination. After the dissection of medaka testis, the sperms can be stored in liquid nitrogen for long periods and used for breeding purposes. In addition, I could get insights into a germ cell transplantation technique for medaka. Further on, I got the chance to join Prof. Naruse and Assistant Prof. Ansai for a memorable fieldtrip, in which we were able to catch wild medaka within different habitats. It was such a great and unique experience for me to see medaka in their natural environment.
Besides getting new exciting scientific insights and further develop my practical skills in the lab, I got to know the fascinating Japanese culture. Experiencing the cheerful atmosphere at several summer festivals with impressive fireworks and traditional Japanese dances and music, visiting breath-taking shrines, castles and temples and falling in love with the extraordinary and tasteful food was stunning. Getting in touch with all of these amazed me more than once.
All in all, I had a great time here in Japan. Both from the research point of view, as well as from a personal perspective, I can take home a lot. I am really thankful for the opportunity to work and life in Japan and highly recommend the NIBB internship program. My personal thanks go to Prof. Naruse who welcomed me warmly in his lab and never got tired in teaching me inside or outside the lab. Moreover, I would like to thank Assistant Prof. Ansai for the inspiring discussions and his support for experimental set ups. Last but not least my sincere thanks go to the rest of the staff, especially to Naoko Torii, Ikuyo Hara, Azusa Yangawa and Toko Yamazaki for the always friendly and helpful manner. Arigatou gozaimasu!
Waseda University, Japan
My name is Chang Yong Park, a 4th year student at Waseda University. During the summer of 2019, I had the privilege to be a part of stem cell laboratory at NIBB as an intern under the supervision of Prof. Tsubouchi. Even though I could be there only for a week, the work experience I got from Prof. Tsubouchi and her lab members was far more than I imagined.
In her stem cell laboratory, I had five major experiments I was involved in : Investigating duration of cell cycle and S-phase length(with Dox and without Dox), E.coli competent cell preparation/check, Sera Lot check (ES handling), ES cloning, and Lymphocyte cloning. On the First day, I was very worried I would not be able to follow the procedures of the experiments, for I never had any lab experiences before. However, thanks to careful supervision and kind guidance of Prof. Tsubouchi and her lab members, I quickly got used to it.
Despite the shortage of time, I was really happy that I could actually get meaningful results from what I did in Prof. Tsubouchi’s lab. This was the first time I was in a big project in science and I am grateful that Prof. Tsubouchi and her lab members provided me a very pleasant first unforgettable experience in a laboratory.
The lodge I was in very pretty wide and clean, and the city Okazaki was peaceful. I did not have any major difficulties while I was there.
University of Washington, USA
My name is Eichi Toyoizumi, a recent Molecular Biology graduate from University of Washington, Seattle and a current intern at Fred Hutchinson Cancer Research Center. I took part in NIBB internship program under the supervision of Dr. Watanabe from Laboratory for Neurophysiology, where he and his colleagues pursue research on animal behavior and human vision using VR and deep neural network.
I was very much intrigued by their research on visual system using deep neural network, however as I have mentioned I was a biology major with limited experience in coding. So my intention was to be exposed in the field of artificial neural network research and to translate the experiences into my future research. Due to some scheduling difficulties my internship duration was little over a week, but it was more than enough to start the research.
I was able to run/program deep learning algorithms to read the sequence of images that I created and make some observations on how they are perceived within the network. But really what amazed me the most is the fact that there is a web-based platform that enables those research to be done remotely allowing for long distance collaboration without one’s presence.
I would like to thank NIBB internship program for the support and Watanabe Lab members: Dr. Watanabe, Dr. Nishiumi and Dr. Kobayashi not only for assistance in research but for all the interesting and valuable discussions. My experience here at NIBB was very positive and will help me through my future research efforts. I look forward to our future, remote for now, collaborations.
To prospective students, I can whole heartedly endorse NIBB internship program to any of you (who are interested in life sciences of course). One piece of advise is that I encourage you to contact the faculty of your interest even if they are not listed as a lab accepting students (my lab was not listed). NIBB has great facilities that equals top research institutions in U.S. such as my alma mater, and so you are in for a treat should you be select to intern.
Vietnam National University HCMC, Vietnam
My name is Nguyen Hoang Yen Nhi. I graduated from the Biotechnology department of International University – Vietnam National University HCMC. Thanks to these two months of NIBB internship at Professor Jun Minagawa’s laboratory of Division of Environmental Photobiology, I found that I was very lucky to have both Professor Minagawa to be my supervisor and Konomi-san to be my mentor. Despite my good Biotechnology background, there were some new techniques for me to learn: sequencing, Western blot, and electroporation. I sincerely thank Konomi-San for her help and guidance. Regarding my project, I used the in-vitro mutagenesis and CRISPR-Cas9 system to create a mutant strains of Chlamydomonas reinhardtii.
During these two months of stay, not only have I just gained more scientific experience but also the chance to visit many places in Japan every weekend to see how beautiful and fascinating Japan is. I am very grateful to NIBB for granting me this opportunity. This invaluable research experience, once again, confirmed my determination to pursue a Ph.D. in algal biotechnology. After this trip, I have become a more mature person with strengthened knowledge and inspiration. In addition to this, I met some great friends that treated me like a family member that I would never forget. I really appreciate their kindness. NIBB internship program is a great opportunity for any research-oriented student to find the path that leads to important career opportunities. The experience and relationships I had during my stay are very precious to me and will be the step stones for me into the scientific world.
Vietnam National University - University of Science, Vietnam
I am Le Thu Hang, graduated student from Vietnam National University - University of Science. Thanks for NIBB internship program, I have got a chance to work at Division of Chromatin Regulation in my three-month internship. During my internship, I had learned a lot through three small project.
The NIBB internship program is a precious opportunity to me. Thanks for the program, I was able to experience a professional research environment as well as Japan culture. Three months spending in Japan is a great time that I will not forget. Friendship and memories come to me unexpectedly, which I bet I will miss a lot when leaving.
I was led to dig into new concepts, to broaden my scientific view in a professional and supportive environment. I would like to send my sincere thank to all members in Nakayama Lab, to my dear friends and mentors for your kind, friendly help, for giving me a warm welcome since the very first moment I came. Please let me express my gratefulness to Professor Nakayama, who has always been patient with my mistakes and taking care of me during my stay.
The internship brings me motivation and confidence to my future plan. Thank you a lot for the opportunity and I hope that in the near future, I could visit NIBB again.
Kasetsart University, Thailand
My name is Lueacha Tabtimmai. I am a Ph.D. student in biochemistry at Kasetsart University, Thailand. I am passionate about scientific research. I think that a little change in science will contribute to a better world. I have a dream about being a part of world-class research group. Finally, I accomplished my goal by being an internship student in NIBB program for 3 months. It is such a fabulous opportunity to challenge myself for getting an amazing experience here.
Under the supervision of Professor Aoki, he is an open-minded person. We first shared the works that we have done so far for getting an idea. I got a chance to create my own project whilst he always shaped the idea and gave me a comment.
Without Aoki-sensei and lab members, I would not have amazing learning. I gain so many valuable experiences during staying in Aoki-lab. I am thankful for all you guys that always give me a ton of warm heart moment. We shared so many cultural things together every lunchtime. Besides from hard-working culture, there are so many activities here that make me really enjoy every day. Onada-san, is a lab secretary who always gives me a hand when I got in trouble. I really appreciate all help from everyone that never denies any inquires. Overall, NIBB gives me the best opportunity for being a part of Aoki lab. I definitely recommend NIBB program for all students who would like to get a good experience in Japan.
Mahidol University, Thailand
My name is Neen Phan-udom. I am a third-year undergraduate student from Mahidol University Thailand, majoring in Bioresources and Environmental Biology (International Program). I had the opportunity to do a 2-week internship at Professor Ueda’s lab- the Division of Cellular Dynamics- which works on plant membrane trafficking proteins in Arabidopsis thaliana and Marchantia polymorpha.
I was under the supervision of Asst. Prof. Ebine during my internship. My work mainly consisted of performing transient assays for Arabidopsis thaliana leaves and visualizing the localization of the targeted proteins via confocal microscopy. I learned to find and take pictures of the cells expressing GFP/RFP-tagged proteins to later measure their activities and possible interactions. I also got to attend a 2-day summer practical course on oil bodies in liverwort which included mini fieldwork to collect liverwort samples and visualize their oil bodies under the microscope.
In addition to gaining more knowledge and laboratory skills, this internship was one of the most wonderful experiences I’ve ever had. I received a very warm welcome from all the lab members as well as other staff at NIBB. I was pleasantly surprised when the lab organized a welcoming party for me and my lab partner on our first day (which has gotten me really into soumen). Everyone was so nice and friendly which made me feel very welcomed and look forward to my tasks every day. I was on the verge of crying on my last day here when I had to say goodbye. I also had many wonderful moments learning about Japanese culture both inside and outside of the lab, not to mention having the privilege to live in Okazaki city and do some traveling on the weekends. Japan really is a beautiful country.
I will never forget my two weeks here. If anything, I wish I could have stayed longer and learned more about membrane trafficking in plants. The internship has shown me the beauty of basic research of exploring the unknowns and left me wanting for more. Really, I can’t thank everyone enough for this amazing experience. I am forever grateful for this internship opportunity.
Visualization of GFP and RFP of targeted membrane trafficking proteins
Welcome party on my first day at the lab
Me and Okubo-san, the lab secretary, who always helped me in everything
Me and Khing performing experiments under Ebine-san supervision
Kyushu University, Japan
NIBB internship program was introduced to me by my teacher, and little did I know, the internship was far more memorable and useful than I expected. Through the internship, I got a chance to take part in current research as well as to experience the life of a research student.
The laboratory that I joined is the Stem Cell Laboratory. Under the instruction of professor Tsubouchi, I participated in two on-going pieces of research: Embryonic stem cell S-phase length measurement and dNTP-recognize complementary protein development. From the experiment, I received many opportunities to practice basic cellular techniques, such as complement protein, cell cultivation, colony identification, etc., that I learned previously.
Another memorable aspect of the internship was experiencing research student life. Every day, I went to the lab at a set time. Initially, professor Tsubouchi described the tasks we tried to accomplish on that day and lectured about the ground understanding of the subject matter, then we proceeded with the job, and after we finished all the experiment on that day, we had small seminars with other lab members. During the small lectures and the seminars, I felt intrigued by the discussion, questions, and answers, and these were the times when I studied new things about stem cells. On some occasions, I needed to be in the lab at night to prepare for the long, time-sensitive experiment. On certain days, the task was only analyzing live-imaging videos of cells, which is far more demanding than it sounds. I must say the life of a researcher is not an easy one. It is difficult and arduous, yet, the moment when I received the expected result, it was very rewarding and worth the effort.
From this experience, I have learned many new things. I have studied the technical knowledge, practiced the skills related to cellular biology, experienced the life of a researcher as I mentioned. More than that, I consider my professor to be a role model of a researcher, being someone who is well organized, dedicate, understands the study thoroughly, and passionate about science. After the internship, I can confidently say that I want to pursue an academic career, and I believe that I can do so.
I want to express my gratitude to professor Tsubouchi and other lab members, the staff members of the dormitory and the institute. Everyone contributes to my highly enjoyable experience at NIBB.
Kyushu University, Japan
Report on Internship at Division of Cellular Dynamics (21 August – 13 September 2019)
This internship was an invaluable and memorable experience. I learned and practiced in several experiments on protein detection using fluorescence microscope and immunoblotting. It was the first time to observe my own plant protoplast samples and scanning with fluorescence microscope under supervision of experts.
Since the first day of arrival, I received a warm welcome from everyone in the laboratory including the supportive staff. It was my honoured to attend Professor Ueda’s lecture on current research focus—genes that responsible for Golgi network transportation in Arabidopsis and oil bodies in liverwort. Afterwards, I was surprised about welcome party and hospitality in the conversation.
I learned a lot of experimental skills from the very essential thing like keeping the protoplast alive, protein extraction, SDS-PAGE, immunoblotting and how to use fluorescence microscope. More than that, the first hand experience from being trained by Assistant Professors of the laboratory was something I could not find from somewhere else. I would like to give special gratitude to Professor Ueda who accepted my application and Assistant Professor Ebine who was my supervisor to most of my experiment and was skillful at troubleshooting the problems. I felt thankful to meet Assistant Professor Kanazawa for his summer school program, Mrs. Okubo for her kindness as a secretary of the laboratory, Mr. Minamino who possess fluent English skill and humour, Mr. Hatchinoda and Mr. Norizuki for his hospitality. I also wanted to thank other few laboratory members that I hardly talk to them for their kind gestures. Last but not least, thank to Mr. Glen, the supporter staff, I received guidance about NIBB and some advice how to adapt myself to Japan.
Szent Istvan University, Hungary
I am Sarah Shaqiri, from Kosovo. I graduated this year Masters of Agricultural Biotechnology in Hungary. As a student I was fascinated with the genome sequencing which we’ve studied in theory quite much, but I was lacking the practice. Fortunately I found NIBB internship and applied in laboratory of Functional Genomics Facility, under the supervision of Prof. Shigenobu Shuji which helped me answer my many questions.
The Shigenobu Sensei’s lab has been studied the symbiosis between aphid and the endosymbiont Buchnera aphidicola. They’ve published several papers on transcriptome of aphid Buchnera and being part of it for a period of almost two months I had to compare the transcriptome between different aphid morphs. It was quite interesting topic yet challenging experience for me because previously I worked on plants and there I was going to work on aphids particularly doing the Next Generation Sequencing for the first time. The project included both computational analysis (bioinformatics) as well as molecular work (bench work).
During the molecular work I was working together with Dr. Nozaki a post doc of the lab. Here I was able to conduct a range molecular experiments like RNA extraction, RNA quality evaluation, RNA library preparation, Real-Time Quantitative Reverse Transcription PCR, tagmentation and generating sequence library using Nextera XT DNA library kit [Illumina], from which we successfully passed this part of experiment. Heading further into computational analysis I faced difficulties but I would want to thank Shunta Yorimoto PhD Fellowship in our lab which helped me giving the best explanations so that I can work further on in bioinformatics in life. I also want to express my graditute to Shigenobu sensei which guided me to be familiar with Unix commands, R commands, DGE-analysis and more. Despite the fact that Sensei had a busy schedule he always found time to help me and give advices to me.
Taken together I had a wonderful experience at the NIBB and in Japan in general. Apart from being welcomed from people in the lab I also made new friends and adjusted myself in the whole different and unique culture of Japan. I am glad I came to Japan and I believe there is much more from it that I could explore. In the end I hope a lot of scientists would have the opportunity to gain experiences like this in the future. It was worth every second.
University of Hawaii at Manoa, USA
Olá! My name is Vânia and I am a 4th year PhD student in the Developmental and Reproductive Biology Graduate Program, Department of Anatomy, Biochemistry and Physiology at the University of Hawaii School of Medicine. Over July 2019, I had the opportunity to do a NIBB Internship under supervision of Professor Toshihiko Fujimori (Division of Embryology).
Over the period of two weeks at NIBB, I was able to enhance my knowledge regarding histological techniques complemented by software analysis that will be used to reconstruct the Astyanax brain in 3D.
Professor Fujimori and Oka Sanae’s expertise and experience alongside with everyone in the lab were crucial for the progress of this project. All lab members were very receptive and helpful which created a very friendly and inviting but also prolific atmosphere. This experience contributed immensely to my project and my personal growth as a scientist, making me more motivated to learn and explore other perspectives.
Overall, NIBB Internship allowed me to experience a new scientific environment and open my mind to future perspectives in Japan and I will always be grateful for the amazing opportunity.
University of Santo Tomas, Philippines
My name is Sheena Josol and I am a Biochemistry graduate from the University of Santo Tomas, Philippines. As someone who had just earned her Bachelors degree, I admit to having limited knowledge which I believed is meant to be enhanced. Thus, it is incredibly fulfilling to be selected on this Research internship program hosted by one of the prestigious research facilities in Japan. I was specifically granted a chance to spend my internship in the Division of Quantitative Biology under the supervision of Professor Aoki.
In my 2 months stay, I was given two research projects which independently aimed to (1) reconstitute tumorigenesis and oncogene addiction in a manner dependent to Trimethoprim and (2) dissect molecular mechanisms of stochastic ERK activation, both of which involved the use of wildtype MCF10A cell lines. In the first project, the oncogene of interest that we intended to induce are the ones involved in ERK signaling pathway:the G protein KRas and the protein kinase BRaf specifically its KRasG12V and BRafV600E mutant version respectively which are widely implicated in several human cancers.
The first step in inducing oncogene addiction was to establish the needed cell lines which in this case is a Dihydrofolate reductase (DHFR)-expressing MCF10A cell line. As we all know, the best way to ascertain the function of one biomolecule is to remove it from the system and see what happens next. The TMP-DHFR system makes it all possible. This model system works in such a way that it requires the protein of interest to be fused to a destabilizing domain like DHFR which targets it for degradation. The protein is then rescued by the addition of TMP that binds the DHFR and inactivates it which in turn enables it to regulate the stability of protein in a rapid, reversible and tunable manner.
Through imaging, western blotting and cell proliferation assay which I spontaneously did during the course of my internship, I was able to analyze the stabilizing effect induced by TMP, quantify the expression levels of induced oncogenes and examine whether our selected cell line acquire cancer like phenotypes after oncogene induction respectively. As for my second project, due to limited time, I was not able to achieve the main goal of studying the stochastic ERK activations.
I am beyond grateful to every member of the lab, who humbly imparted their knowledge to me especially to Aoki-sensei and to my mentor Reina-san who patiently taught and guided me with everything. I would also like to express my gratitude to Onoda-san, the secretary in our lab, whose unfailing efforts made my first ever Autumn experience the best. I really appreciate the good laugh I’ve shared with everyone during lunch time and all the parties they have prepared just for me.
Overall, this experience has not only filled me with significant learnings in the field of science but also let me gain great memories, learn meaningful life wisdom, establish lasting friendship with people from different parts of the world and lastly brought me to wonderful places that made me appreciate the beauty of Japan more. Indeed, Japan is a beautiful country but more than everything else, I think the culture of hardworking, disciplined and kind people makes it more special.
Mahidol University, Thailand
My name is Watcharin Unwet, a 4th year undergraduate student from the Department of Biology, Faculty of Science, Mahidol University, Thailand. I had an opportunity to join the laboratory of plant development and physiology led by Associate Professor KAWADE, Kensuke. This laboratory studies how metabolic systems are coordinately modulated with developmental progression on plants. The main focus for this particular internship project is to study how to regulate cell proliferation and expansion process of Arabidopsis thaliana.
I had a wonderful time during internship program period. In 15 days, I learned a lot of skills including image analysis using FIJI (A distribution of ImageJ software program which includes many useful plugins contributed by the community) which is a very useful tool for biologist and learned some essential laboratory skills for plant biology such as molecular biology technique, using confocal microscopy to study cellular morphology of plant, using Katikati2 software program for counting the plant epidermal cell and palisade cell and so on. These techniques can be applied to my senior research project in my university.
As a biologist who loves to study plant science, I felt really enjoy working in this lab. I would like to thank all Kawade’s laboratory members: Kawade, Tomoi, Nozaki, and Yamaguchi for everything. I also enjoy my daily life in Japan, I had learned some basic Japanese sentences and their cultures. Moreover, I had met with so many foreign friends there and had a great time together.
I would like to give a big thank to NIBB internship program for making this memorable experiences possible and I would like to encourage those students who interested to study biological science and learn some Japanese culture. For me, this internship program is one of the most valuable moment ever.
Kyushu University, Japan
I am 4th year undergraduate student in Kyushu University, majoring in marine biochemistry. I currently study immune system of fish, focusing on their innate immunity. The reason why applied this course was because I wanted to enhance my understandings in genetics and also learn related experimental techniques in order to apply such knowledge in my future research. I believe understanding genetic background of the target organism is important to take the research to the next depth. Also, knowing genetics approach to tackle the research problem could be powerful in my future research. A week visit in Prof. Naruse’s laboratory of bioresources was very exciting, and I was able to learn basics and applications of genetics research using medaka (Oryzias latipes) as a model.
Medaka is a small freshwater teleost fish that are now developed as a model organism for vertebrate. Given that its genome sequence is now available on database, medaka is ideal for genetics research since they lay eggs in very frequent pace. It is my unforgettable memory to see a breeding sector full of different strains and generations of healthy medaka. Variations in phenotype of different medaka strains were very interesting to compare.
During this week visit in Prof. Naruse’s laboratory, I experienced a set of experiments that aimed to observe two body color mutant phenotypes of medaka to understand genetic linkage and polymorphism by measuring genetic distance based on loci of certain genetic markers. The set of experiment included the sorting of F2 embryos, fin clipping of F1 parents, genomic DNA extraction, PCR and restriction enzyme treatment of PCR product. In addition to these experiments, I also had a chance to experience a brief training on microinjection technique using CRISPR-cas9 method. These sequence of experiments taught me the logics of constructing a convincing analysis.
Moreover, the program was not limited to the experiment. Additional lectures on background knowledge of the experiment, efficient use of database, and on primer designing method are some of the things I learned during this internship. Those practical knowledge I gained from the lectures are now supporting me in my laboratory.
In this extremely short period of visit, I was able to learn countless amount of knowledge from very packed content of the internship. I really appreciate Prof. Naruse, Assistant Prof. Ansai, all laboratory members, and internship staffs for preparing such a wonderful program. It is my honor to say that I have completed an internship at NIBB, especially in this laboratory. Even from this short visit, I can proudly say that I have learned so many valuable things from every experience I had during the internship.
Kyushu University, Japan
My name is Do Thuy Linh and I am a 3rd-year student of International Undergraduate Program, Department of Bioresource and Bioenvironment, Faculty of Agriculture, Kyushu University. During August and September 2018, I was selected to do the internship in the Laboratory of Functional Genomics Facility, under the supervisor of Assoc. Prof. Shigenobu Shuji. During 2 months of internship, despite I am the youngest intern ever (just finished 2nd-year undergrad), he still assigned me a challenging project which is not just a training or a practice the results have been known, but new project nobodies have explored yet. As he said, "so that you can experience the truly scientific project". My project is to identify and analyze the spatial expression of BCR3 and LSZ-i genes in bacteriocytes of the Ceratovacuna japonica. BCR3 and LSZ-i have already identified as specifically expressed in bacteriocytes of the pea aphid, Acyrthosiphon pisum. These genes play important roles to meditate the symbiosis between pea aphid and Buchnera. The project included both computational biology (bioinformatics) work as well as molecular biology (bench work).
From the document provided by Shigenobu sensei and guided by Mr. Shunta Yorimoto, I have learned about UNIX commands and BLAST. After successfully identification of BCR3 and LSZ-i homologs in C.japonica transcriptome, with the enthusiastic and conscientious support from Dr. Chen-yo, who is doing Postdoctoral Fellowships in our lab, I was able to conduct a wide range of molecular experiments from RNA extraction, RNA quality evaluation, Real-Time Quantitative Reverse Transcription PCR, in situ hybridization, using the mRNA transcriptome sequence of bacteriocytes. The result indicated that both BCR3 and LSZ-i show the gene conservation in C.japonica.
There were a lot of other things about this program that were worth being excited about. First of all, the sheer idea of spending my summer in NIBB thrilled me: it’s the centre of research, of state of art technologies, of professional experts and of friendly people. Shigenobu sensei is so supportive and took care of me very much. Although my professor is really busy with so many business trips every week, he still asked me: "Apart from science, would you like to visit some sightseeings in Okazaki?". Then he and his wife took me and Mr. Chenyo to go visit both traditional and modern sightseeing in Nagoya, and also drink coffee and have many talks about life and science careers. Moreover, I also met a lot of interesting people in my program: Miyuzu-san, Bino-san, Ichikawa san and other staffs in the Common Facilities. They were and are still my inspiration.
After this program, I feel contented that I have got the chance to get to know scientific research in many aspects. Each of the techniques and knowledge acquired in the program will serve as a door to an educational pathway that I may choose to pursue later on during my study at Kyushu University or beyond. In the end, I felt so honored to be selected for the internship in NIBB and Assoc. Prof. Shigenobu Shuji’s lab and I was in the best research environment ever.
Peking University, China
I am Peng Chen, an undergraduate student from School of Life Sciences in Peking University. In the last summer vacation in my college life, I was fortune to take part in the NIBB Internship. I felt honored to work in Professor Hasebe’s Lab and really enjoyed an impressive life in those two months.
During my stay, I did some researches on auxin biosynthesis in the moss Physcomitrella patens (Physcomitrella). The experimental material was a mutant, showing defects in periclinal cell divisions, which is one of the most significant developmental process for land plants.
I must express the depth of my gratitude to Professor Mitsuyasu Hasebe for giving me the opportunity to learn and work in his lab. He is an erudite scholar and a kind supervisor for students. And I should show my appreciation to Dr. Tsuyoshi Aoyama who guided my researches in those two months. He answered every question I asked patiently and taught me a lot about Physcomitrella and its experiments. Also, many thanks to all the members in Professor Hasebe’s Lab. You were glad to give me a favor every day and showed me an outstanding, efficient and optimistic scientific research team. Specially, I would like to say thank you to Mr. Cowan in international cooperation office, for his responsible attitude towards all the procedures during my intern and his concerns for my daily life.
This was my first time being aboard, but I felt at home in Okazaki. I love this quiet and peaceful city and I think it is one of the best places for scholars to study their scientific topics. I spent an unforgettable time in NIBB and I hope that I can continue my graduate education in Professor Hasebe’s Lab.
Atma Jaya Yogyakarta University, Indonesia
Halo ! My name is Astrid Valerie Putri Kusuma. I am a biotechnology student for undergraduate study in Atma Jaya Yogyakarta University, Indonesia. I had the privilege to do a brief internship for 4 weeks at Division of Chromatin Regulation which was led by Nakayama Sensei.
I worked on the project for characterization of HP-1 like protein in Tetrahymena. HP1 is a conserved chromosomal protein that forming higher order of chromatin structures and binds to H3k9me (lysine 9-methylated histone H3). HP1 proteins have an amino terminal CD and a carboxyl-terminal CSD. There are more than 20 known heterochromatin protein but we still do not know how they interact with each other.
Therefore, the tasks that was given to me is to construct plasmids for expressing 12 CDs in Tetrahymena HP-1 like proteins. These will be used to obtain 12 CDs in Tetrahymena HP-1 like proteins and the final goal is to analyze chromodomain’s affinity for H3K9me3 or H3K27me3 by using isothermal titration calorimetry.
My internship so far is extraordinary. I am truly honored and blessed to work under Nakayama Sensei. He is such a great person and always willing to help and assist me in doing my project. This kind of humble treatment is one in a million and is not famous in my country. The other staff members are also really kind. Thanks a lot to Ken, Aki, Asai, Yuriko, and Kyoko for being supportive and patient with me. All of them were really helpful in helping me working in the lab and for any other administration.
I also feel grateful for my accomodations. Although my arrival was planned in such a short time, but they manage to deal with all of it. Special thanks to Glen who took care with the administrations regarding my arrival and accomodations. The people here are so nice and it’s really easy to make friends.
Last but not least, this is one of the great experience that I will always remember. Though it sounds exaggerating but when treatment with kindness is almost in every part of the process, this will make someone always remember. I hope that I will be given the chance to be back here someday. Terima kasih banyak !
Hanoi University of Science and Technology, Viet Nam
My name is Nguyen Ha Ngoc Anh, I am a fresh graduate from Hanoi University of Science and Technology, major in Bioengineering. During the spring of 2019, I was lucky enough to take part in the NIBB Internship under the Division of Environmental Photobiology, with supervision from Prof. Jun Minagawa and mentor Ryoichi Satou. It was a remarkable experience in regards of both academic and life style-wise.
My research revolves around lutein production in microalgae. Lutein is a valuable biocompound, playing a major role in eye protection and hold a promising future in the food supplement industry. This is a long on-going research and I took multiple approaches towards boosting lutein accumulation in Chlorella sorokiniana, regarding from strain selection, media optimization to mechanism verification.
Despite the lack of time, this journey has plenty take away lessons that I hold at heart. First and foremost, I would like to express my gratitude to Minagawa-sensei, my mentor Satou-san and all lab members for a wonderful 3 months. Not only did they welcomed me with the warmest hearts, they also introduced me to a nurturing science community, where lab members can deliberately discuss and groom each other to success. Even though I am still young and rather inexperienced, all lab members always come to my aid, give me advices to help me freely test out my vision, provide me the opportunity to take multiple approaches in the matter. Furthermore, during this trip, I was able to make invaluable friendship with people from all around the world, who all had inspired me in every way possible. The spring of 2019, filled with sakura, pot-luck parties and laughter will always be the memory that I treasure.
In conclusion, the NIBB Internship is a remarkable, high-quality opportunity for all biology students who wishes to embark on a scientific career. I would like to say thank you to everyone at NIBB who had made this journey possible for me and I wish everyone the best of luck in the future.
University of Belgrade, Serbia
My name is Olivera Valentirović and I am an undergraduate student at Faculty of Biology at the University of Belgrade, Serbia. Thanks to prof. Dr Nakayama who suggested the NIBB internship program to me, I was able to experience working at his laboratory for one month. It was a great adventure and opportunity for me, since I had a chance to work in the field of epigenetics and to travel to a breath-taking far away country, that was something I only dreamt about before .
The research I performed during my internship at NIBB was on the ciliate Tetrahymena thermophila. This was my first time working in a laboratory as well as with Tetrahymena. I found its life cycle and the extreme processes happening on its genome during its sexual reproduction process called conjugation extremely fascinating. Those peculiarities make it a really favorable and attractive model organism.
My project was to characterize the proteins involved in the formation of heterochromatin and DNA elimination during genome rearrangement within the macronucleus. In one experiment I generated several transgenic cell lines that express proteins tagged with green fluorescent protein to analyze localizations of heterochromatin protein candidates. It was a great experience seeing gene gun transformation for the first time, and then learning how to use it myself. I also learned a lot about culturing, storing and transferring Tetrahymena and how to perform a selection of the transformed cells and phenotype assortments using different concentrations of selective substances in the medium. In a parallel experiment, I aimed to reveal the sufficiency of few proteins for DNA elimination by a tethering assay. I faced some difficulties during this experiment, but I found it a good experience for my future work, because that I may face and need to solve on a daily basis. Every day I was presented with interesting challenges and new chances to find a way to untangle some problem. In the end, I managed to go through all the intended steps and successfully finish the experiments with some of the samples. I wish I could have had a longer period of time to do more experiments with different conditions, and finish all of the initially planned experiments.
Accommodation at Myodaiji Lodge was extremely comfortable which made my stay very pleasant. I didn’t lack anything and everything was professionally organized, thanks to the International Corporation Office.
All of the lab members of the Division of Chromatin Regulation were always willing to help me with my work, as well as with my stay. They gave me a lot of precious advices for which I am immensely grateful. It is a great honor for me to have worked with such polite, kind and hard-working people. I would especially like to thank prof. Dr Nakayama who gave me this opportunity to join his lab and offered me an unforgettable experience of Japanese culture, its delicious cuisine and its people. I am eternally thankful to Dr Kataoka from whom I’ve learned many valuable things, who patiently and carefully watched and guided me through every step and who showed me how wonderful and incredibly fun working in the laboratory can be. I really appreciate friendly and welcoming atmosphere during my stay.
This was a life-changing adventure for me. In addition to the exciting bench work in the actual lab, I’ve gained many unforgettable memories, visited amazing places, felt one unique culture, met remarkable people and made lasting friendships. The whole experience inspired me to pursue my career in science even more than before.
Mulawarman University, Indonesia
My name is Anisa Fitri Rahayu from Indonesia. Recently, I have graduated from Biology Department, Mulawarman University. I was doing internship about 1 month at Prof. Nakayama's Laboratory. I am so curious about the regulation of gene expression, especially how cells up- or down-regulate genes without changing the DNA sequence. This was the reasons why I applied to this NIBB internship program. During my internship, I worked with fission yeast Schizosaccharomyces pombe (S. pombe), and studied the factors that play an important role in heterochromatin stabilization and silencing.
In fission yeast, such as S. pombe, there are two HP1 family proteins (Swi6 and Chp2) and play as main factor for maintenance gene silencing at heterochromatin regions. These HP1 proteins recruit other binding partners in order to maintain the repressive structure of heterochromatin. The exact means by which such HP1 family proteins recruit distinct set of partners, however, remain incompletely understood. My study in this internship program consisted of disturbing the function of one of Swi6-interacting proteins and characterizing mutant strains by silencing assay. The effect on heterochromatin integrity and stabilization can be check by growing the cells in medium containing low level of adenine, because on this study I used the S. pombe strain carrying ade6+ marker gene in the heterochromatin region on centromere 1. The isolated cells showed variegated phenotype on the medium resulting in white and pink colonies and indicated that silent chromatin at centromere is less stable in the mutant strains.
I feel lucky got the opportunity to join the program because I got some precious experiences not only about lab work but also about Japanese culture. I had chance to attend the laboratories join meeting such as paper and progress research presentation also watched seminar from other laboratories. All the knowledge I got there was new and could answer my curiosity about heterochromatin also made me understand some concepts that I had not understood before. I hope the experience and knowledge I got from the program can help me to pursue my career plan as researcher in the future. I got nice atmosphere too in the lab and I really enjoy my time there, I would like to thank for all laboratory members: Machika, Naoko, Kyoko, Yuriko and Ken for helped me in the lab and being friendly, especially for Nakayama sensei for the guidance in lab work also helped me understand all about epigenetics, heterochromatin as well as helped me to prepare my entrance examination to enter NIBB graduate program (SOKENDAI). I hope to have opportunity in near future to come back and study at NIBB.
Pontificia Universidad Javeriana in Bogotá, Colombia
Phylogenomic studies showed that in Colombia a new subpopulation of H. pylori has evolved. During this training a computational pipeline to inferring the ancestry of this subpopulation was developed. Next, a brief description of the pipeline steps is presented.
1- MBGD: orthologous detection.
MBGD is a database system developed to compare different features of bacterial genomes, such as: gene order, motif identification and orthologous/paralogous detection. I have received training about how to use not only MBGD but also the RECOG tool to extract the core genome orthologous alignment from 141 H. pylori genomes.
2- SNP calling and the vcf format
The full genome and the concatenated core genome sequences were used to create vcf files for each strain using the snippy tool. Finally, the vcf files were merged independently using vcftools.
3- Imputation and fineStructure analysis
Each merged vcf file was imputed using BEAGLE and the fineStructure software was used to obtain the ancestry matrix for this subpopulation.
4- Positive selection analysis
Finally a whole genome positive selection analysis was conducted using PhaME.
5- Scripting
Professor Uchiyama teaches me how to write scripts in order to automate all the analysis using bash shell.
About my experience in Japan
Japan is a beautiful country full of kind, humble and educated people. I am just amazed about how good is this precious country. Respect to Okazaki, the city is quiet and extremely safe. Its mood is friendly, sweet and everything is close to you. So, it is possible to enjoy a delightful walk seeing amazing houses, trees and landscapes just going to the grocery. The food in this city is fantastic, is the best food I have ever tasted. Finally, I was hosted at the Michima lodge. This place is cozy, silent and friendly. I have felt like in home. I want to thanks Abe-san for this.
My internship was directed by Uchiyama sensei. He is not only one of the best bioinformatician in the world, but also one of the best human beings I have ever met. From him, I have learned extensively not only about genomic but also how to be a better person. I also want to thanks the Office of International Cooperation specifically Ritsue Takahashi because she was essential before, during and after the internship program. In general, the NIBB is the best academic place I've ever known in my entire life. Here, you can learn how to do the best science. I hope I can return one day to this incredible institute.
Kyushu University, Japan
My name is Thi and I am currently a 4th year undergraduate student majoring in Applied Biosciences at Kyushu University. During the two-week internship at the Laboratory of Stem Cell Biology headed by Prof. Dr. Tomomi Tsubouchi, I was very lucky to assist all lab members in their ongoing projects revolving around genome stability mechanisms of embryonic stem cells (ESCs) and their roles in reprogramming and differentiation.
ESCs have a unique cell cycle of shortened gap phases compared to other cell types. Analyzing the differences in DNA replication (S phase) and mitosis (M phase) of ESCs and differentiated cells prove a useful approach to elucidate how and why ESCs proliferate with such unique cell cycle. In one experiment, I assisted in sorting ESCs and differentiated fibroblast cells at different S phases, from which samples will be used for further analysis in their replication process.
Despite the short time, thanks to the kind instruction of all the lab members, I was able to learn a wide scope of laboratory techniques such as cell sorting, FACS analysis, confocal microscopy, live imaging analysis, tissue culture handling of various cell lines.
I would like to express my greatest gratitude for Tsubouchi-sensei and all the members of Laboratory of Stem Cell Biology for their very kind guidance, delightful conversations on Kansai culture, scientist life stories and the wonderful misokatsu cuisine. Apart of lab work, the wonderful accommodation arrangement at Mishima lodge has made my stay very comfortable and helped me make unforgettable friends.
At last, I am eternally grateful for NIBB to give me the opportunities to work alongside with such hardworking, kind and talented scientists in the Laboratory of Stem Cell Biology. The research atmosphere here is truly inspiring and exciting and I really hope to come back here in the future.
Universiti Sains, Malaysia
I am Kathrine Tan Xin Yee, a graduate student from Universiti Sains Malaysia. I am truly grateful that I could take part in this internship program. I was assigned under the supervision of Assoc. Prof Shuji Shigenobu and I am impressed by his passion towards research study. I feel honored to be able to work with him and also his lab members.
I involved in the project entitled ‘ Identification and Quantification of Symbiotic Bacteria in a Social Aphid, Ceratovacuna japonica by metagenomic approach’. C. japonica is a species of eusocial aphid. One of the distinctive features of this aphid is the evolvement of soldier caste in the colony. This unique feature has distinguished C. japonica from the model species, Acyrthosiphon pisum and it is possible that the symbiotic bacteria presence in the soldier caste and reproductive caste of C. japonica as well as the A. pisum are different.
Throughout the three weeks of internship, I tried to identify and quantify the secondary symbionts that present in the reproductive caste of C. japonica. The first week of my internship was spent in conducting laboratory works such as extracting genomic DNA from the reproductive caste of C. japonica. Preparation of 16S metagenomic sequencing library was conducted in the second week followed by sequencing of metagenomic library using Illumina Miseq sequencer. I would like to thank Ms. Miyuzu Suzuki and Ms. Asaka Akita for assisting me in the experiment. While waiting for the results of sequencing, I spent most of my time in preparing the workflow for the analysis of metagenomics data. As I am not familiar with writing scripts and UNIX commands, therefore I spent quite a considerable time in learning how to operate data analysis pipelines like QIIME. However, with the help of my dearest lab member, Mr. Shunta Yorimoto, I was able to perform simple data analysis and certain data processing commands. I am glad that the analysis demonstrated the possibility of finding novel symbiotic bacteria in C. japonica.
Apart from that, I am thankful to the members of NIBB Core Research Facilities, Functional Genomics Facility Department. Without them, I do not think that I could adapt to this new environment in such a short time.
Terima kasih andありがとうございました.
Heidelberg University, Germany
I am currently enrolled in the master program “Molecular Biosciences” at the University of Heidelberg in Germany. Studying in the major program “Developmental and Stem Cell Biology “, I am mostly interested in processes that determine animal development from a simple egg to a highly complex organism.
Already during my bachelor thesis in Heidelberg, I worked with the Japanese rice fish also known as medaka. Proven as a highly suitable model for transgenesis experiments, I wanted to expand my knowledge about this sweet water fish and improve my experimental techniques in the laboratory. Therefore, I applied for an internship in the laboratory of Prof. Kiyoshi Naruse studying the genetics underlying pigment cell and neural development in medaka.
During my 2-months stay at the NIBB in Okazaki, I focused on the development of a moto neuron reporter line by using CRISPR-Cas9 mediated knock-in of a green fluorescent protein. Therefore, I designed sgRNAs targeting the 5’UTR of neural genes that are exclusively expressed in the brain. These sgRNAs were conjected with Cas9 mRNA, a donor plasmid containing a Tbait sequence and a Tbait sgRNA to open the donor vector. Unfortunately, the chosen genes seemed to have an essential role in embryo development, so most of the injected embryos did not survive until hatching. I optimized the system by targeting more sequences 600-200bp upstream of the transcription start side and by adjusting donor plasmid and sgRNA concentrations. Luckily, promising neuronal GFP expression could be found at last. Additionally, I used a donor vector with a DsRed flanked by LoxP sites followed by GFP. Upon Cre recombination i.e. by crossing to a Cre expressing line, DsRed will be excised leading to GFP expression. The injected fish with mosaic fluorophore insertion are now growing and will be tested for exclusive moto neuron expression in the next generation. Eventually, they will become founder of an inducible moto neuron transporter line.
Facing obstacles, I was obliged to think about how to design and conduct experiments to prove hypotheses and optimize the experimental procedure. Working independently on my project helped me to gain self-confidence and comprehension of scientific research. Hence, I am very grateful that Prof. Kiyoshi Naruse gave me the opportunity to work in his laboratory. I would also like to thank Satoshi Ansai who was always there to answer my questions and all the lab members who helped me to find my way in the lab. Special thanks to Chieko-san for explaining me about Japanese language and culture during lunch breaks. Finally, I would like to thank the NIBB internship program for its financial support.
My stay in Okazaki was both enriching and inspiring thanks to all the people I met on the way. Coming to Asia for the first time in my life, certainly changed my thinking and understanding. I am very glad that I came to Japan and hope that a lot of students will benefit from this program in the future.
Heidelberg University, Germany
I am a developmental biology MSc student from Heidelberg University and in fall 2017, I had the opportunity to join the group of Prof. Kiyoshi Naruse under the NIBB internship program. During my stay, I was involved in a research project on animal pigment cell and body colour evolution.
Prof. Naruse’s laboratory of Bioresources provides access to numerous medaka (Oryzias latipes) strains as well as a plethora of other Oryzias subspecies. While closely related, these fish have, for instance, not only exhibit disparate mechanisms of sex determination but also distinct pigment cell (chromatophore) type occurrence. Thus, they constitute suitable model systems for studying evolutionary processes.
Whereas higher mammals feature only one chromatophore type (melanocytes), several are found in teleosts (e.g. black melanophores, yellow xanthophores, blue cyanophores). Another variant, the orange-white leucophores, are present only in a few fish species. They are found in medaka, however not in O. woworae, an indonesian relative. Yet, intriguingly, the genes underlying leucophore development and specification in O. latipes are conserved in the latter. This hence raises the question of how these genes have functionally evolved in O. woworae.
To tackle the aforementioned conundrum, we employed the CRISPR/Cas9 system to knock-out and label two of those genes, pax7a and slc2a15b, in both species. Pax7a is of particular interest, as it is also crucial for xanthophore development in medaka. Ultimately, several transgenic O. woworae and O. latipes lines were generated for each of the genes listed above. These either simply lack functional target genes or harbour a GFP coarsely inserted into the endogenous loci, resulting in a disruption of their respective ORF, i.e. a knock-out via fluorescent reporter knock-in, which, in turn, further enables the study of target protein localization and gene expression dynamics through life-imaging.
As to my everyday life outside of science, everything provided by the administrative office was very convenient and I felt comfortable living in Okazaki. On the weekends, I was able to explore other cities and parts of Japan which also left me with many fun and memorable impressions.
Taken together, I had a wonderful, instructive experience at the NIBB and in Japan in general. Accordingly, I highly recommend the NIBB internship program. Every member of the research group (and all other staff I met) were extremely welcoming and friendly. Hence I would like to thank each one, as well as specifically Dr. Satoshi Ansai and those who assisted me during my project. I would further like to express my gratitude to Prof. Naruse for being a very kind and hospitable host who generously invited me to join a national scientific meeting and warmly introduced me to several aspects of Japanese culture.
Dartmouth College, USA
I am a third-year undergraduate chemistry student at Dartmouth College in the United States, and had the opportunity to spend three months in Professor Aoki’s lab in the Division of Quantitative Biology at NIBB as a research intern.
My research project involved studying various signaling proteins by quantitatively manipulating their intracellular abundancies. This was achieved by using a new experimental tool that involves genetically fusing engineered dihydrofolate reductase (DHFR) to a protein of interest, then introducing a small-molecule DHFR inhibitor called trimethoprim (TMP). In the presence of TMP, the fusion protein is stabilized, but in its absence, it is degraded. This allows for the fine-tuning of the intracellular concentrations of proteins of interest. Using this so-called DHFR-TMP protein stabilization system, I studied the phenotypic effect of three signaling proteins, Rac1 (involved in cell motility), MEK1 (involved in cell proliferation), and BAD (involved in apoptosis induction), by genetically fusing them to DHFR and a green fluorescent protein and tracking their intracellular abundancies by fluorescence microscopy.
As a chemistry major and moreover merely an undergraduate, I was completely unknowledgeable and unfamiliar with the field of life sciences when I first arrived at NIBB. However, through this internship program, I was able to learn about and experience biological research first-hand, and even had the opportunity to present my research in a poster session at the annual retreat of the Okazaki Institute of Integrative Bioscience. This research experience also confirmed my interest in attending graduate school to pursue a PhD. I would like to thank Professor Aoki for giving me the opportunity to join his lab through this internship program and also Miura-san, a graduate student in the lab, for her help and guidance in my research project.
My internship experience in the Aoki lab was not only academically enriching, but also (to put it simply) incredibly fun. I felt welcomed in the lab from day one thanks to Professor Aoki’s humorous personality and the warmth and kindness of all of the lab members who helped me all along the way. We would eat lunch together as a lab every day over hilarious conversation, make tacos and spam musubis together for parties with other labs, go out to eat at Cannery Row for the buffet…I am eternally grateful to everyone in the Aoki lab for making my three months here, however brief, so memorable and so invaluable.
Middle East Technical University, Turkey
My name is Emre M. Ipekoglu. I am an undergraduate student in the Department of Molecular Biology and Genetics,Middle East Technical University, Turkey. I was quite lucky since I have met with Prof. Minagawa at a conference who had given this internship opportunity to me. During my internship period, I had not focused onto a particular subject, rather I tried to learn research subjects of the every member in the laboratory, Division of Environmental Photobiology. It was a fulfilling process for an undergraduate student due to the high number of researchers and various top techniques that are being used in the laboratory. Then, I had the chance of performing experiments related to non-photochemical quenching and genetic screening of the Chlamydomonas reinhardtii. Every discussion with members of the laboratory was a gain for my ability to keep up with the scientific process.
I believe that I soaked up the atmosphere not only for academic aspects, but also for cultural domain which was another important gain for my intellectual development. Also, I tried to introduce my culture, it was a perfect sharing occasion, thanks to this friendly and supporting environment in the laboratory.
The organisation of the internship was very professional thanks to the international cooperation office. Accommodation in the Myodaiji Lodge was comfortable and neat, everything was well-organised in order to meet the every needs of the visitors.
I would like to acknowledge Minagawa-sensei and dear members of his laboratory.
Vietnam National University – University of Science, Vietnam
I am Tran TH Nguyen, from Hanoi, Vietnam. I have just completed my Bachelor’s in Microbiology from Vietnam National University, University of Science. I was so glad to be an NIBB internship student under the guidance of Professor. Takada from 10th Oct to 28th Dec, 2016. During my internship in the molecular and developmental laboratory, I focused on the expression of EGFP fluorescently tagged Wnt3a protein during the embryogenesis of Xenopus embryos.
In my research, I planned to visualize Wnt proteins by the addition of fluorescent tags because it is known to be quite difficult to generate anti-Wnt antibodies available for immunohistochemistry. Prior to visualization of tagged-Wnt proteins in embryos, in the beginning, I focused on the optimization of linker length to minimize the effect of tags on the activities of Wnt proteins. According to some recent research, it was shown that activities of Wnt proteins are frequently damaged by the addition of fluorescent tags. Therefore, I tried to optimize the design of tagged proteins by changing the length of the liker peptide connecting Wnt to EGFP tag. Specifically, I generated 5 constructs in which the lengths varied from 9 to 55 amino acids. These constructs were expressed in culture cells and in Xenopus embryos and examined to what extent Wnt3a activity and EGFP fluorescence were retained.
During my three months of stay in Okazaki, I had lots of unforgettable memories with labmates and other friends here. I was able to learn some interesting research topics in my internship under the valuable guidance and inspiration of Prof. Takada, my mentors – Ritsuko san, Mii san, and the warm hearts of Nobata-san, Utsumi-san, Takashiro-san and other labmates who encouraged me to successfully complete my internship here. During my first visit in Japan, my flight was delayed by 2 hours and I arrived in Nagoya at midnight. It was so touching that Prof.Takada and his wife came to receive me from the airport so late at night.
Furthermore, I can never forget to mention about my stay in Okazaki would never be meaningful without the taking care of my kind-hearted host family. I would like to express my sincere thanks to Fukada-san, Makiko-san and my host brother - Shortaro-kun who always take care about my needs, be with me in many short trips to explore Okazaki, Nagoya… Thank you so much for treating me as your daughter!
Last but not least, many thanks go to my beloved parents, my “someone” for unconditional love and continuous supports me when I went abroad.
The NIBB internship program is really meaningful for international students who want an experience with one of the highest reputations in education as well as the Japanese culture. Thanks all, for everything I experienced here!
University of Delhi, India
I am Preeti Khandelwal from Department of Zoology, University of Delhi, India. I have submitted my PhD thesis in fish endocrinology.
Fish have a variety of pigment cells or chromatophores including melanophores, xanthophores and iridophores. Medaka has a unique chromatophore type called leucophores in addition to above three. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/ CRISPR-associated protein-9 nuclease (Cas) system is the most extensively used genome editing tool, which is essential in adaptive immunity in bacteria and archaea, enabling the organisms to respond to and eliminate invading genetic material. Three types of CRISPR mechanisms have been identified, of which type II, is the most studied. In type II CRISPR system, invading DNA from viruses or plasmids is cut into small fragments and incorporated into a CRISPR locus amidst a series of short repeats (around 20bp). Protospacer adjacent motifs (PAM) 5'-NGG-3' are DNA sequence immediately following the DNA sequence targeted by the Cas9 nuclease in the CRISPR bacterial adaptive immune system. The loci are transcribed, and transcripts are then processed to generate small RNAs (crRNA – CRISPR RNA), which are used to guide effector endonucleases that target invading DNA based on sequence complementarity. The type II CRISPR mechanism is unique because only one Cas protein (Cas9) is required for gene silencing.
The topic for my internship project was “ CRISPR/ Cas mediated gene knockout to generate see-through medaka”. To generate the knockouts we used the CRISPR/Cas based RNA-guided endonuclease in cab strain of medaka. Five causative genes for pigmentation in medaka were selected namely pnp4b, dnajb14, bnc2, pisp1 and snapc3. First step is to prepare cas9 nuclease and engineered single-guide RNAs (sgRNAs). Capped RNA encoding for Cas9 nuclease is transcribed from an expression vector pCS2+hSPCas9, constructed for fish. Specific sgRNA for each above mentioned genes are engineered by cloning a pair of custom-ordered oligonucleotides in vector pDR274 and then in vitro transcribed using T7 RNA polymerase. I successfully completed the key step of my project.
Every morning freshly released fertilized eggs are collected and microinjected the one-celled eggs with the Cas9 nuclease (200ng/µl) and each sg RNA (50ng/µl) mix, using a thin glass needle held in a micromanipulator. The mRNA used is very important in deciding the role in pigmentation in medaka. After incubation of fertilized eggs at 28 ℃ for 3-5 days, genomic DNA is extracted from each egg. The target sequence are amplified by PCR using specific primers for each gene. PCR amplicons are applied to gel electrophoresis on the latest equipment, MUltiNA, which made the gel electrophoresis very simple, fast and easy to analyse. The electrophoretic results are analysed to estimate the targeted genome modifications and identification of mutants by Heteroduplex mobility assay (HMA). In case of discrimination among wild type heterozygous and homozygous, wild type and homozygous mutants show a single band with small difference in their mobility whereas heterozygous mutants exhibits multiple bands. Out of five sg RNAs , four sgRNAs induced mutations.
I really wished to perform further experiments to get the end results of my project but I had only one month which was very short duration to carry out all the experiments. But everyday was a learning day for me. As a fish endocrinology background these experiments were absolutely new to me but I am really grateful to assistant prof. Yusuke Takehana, who really helped me at every step of my experiments and taught me to perform microinjection easily in the medaka eggs, which I was finding very difficult initially. He also provided me the bicycle to commute to the lab from the lodge and to explore the Okazaki. Cycling everyday helped me to stay healthy. I truly appreciate the interactive and friendly atmosphere in NIBB and prof. Naruse’s Lab. All the lab members and staff are very helpful and outstandingly supportive.
I enjoyed my stay at Myodaiji lodge which is very well equipped, organised and comfortable. Its location is so apt which made my transition very smooth. I also got the chance to visit some beautiful tourist places in such as Takeshima island, Okazaki castle, Arashiyama and kiyomizudera temple in Kyoto to understand the Japanese culture. I enjoyed every day in this incredibly beautiful, safe and fascinating country. This internship aid in my overall scientific development and also from cooking my own vegetarian food to exploring rich cultural values of Japan.
I hope to have another opportunity to come back to NIBB in near future and work under the supervision of Prof. Kiyoshi Naruse and Assistant Prof. Yusuke Takehana to gain as much as knowledge from them.
Heidelberg University, Germany
I am currently a M.Sc. student in Germany, Heidelberg in the field of Molecular Biosciences. I had opportunity to experience NIBB internship for nearly 3 months with the help of one of my professors in my department. In Heidelberg, my professor is working on eye development in medaka fish, a Japanese rice fish. My field of study in NIBB was the sex determination in O. hubbsi fish which is a close relative of medaka, O. latipes. Thanks to the collaborative studies between Heidelberg and Okazaki, I learned about the NIBB internship program.
There are several advantages of this internship for my academic career. First of all, O. hubbsi was a new model organism for me to apply genetics and molecular biology methods. Also, sex determination field was a new concept for me to study. In this 3-month internship we were trying to identify a new gene which determines either male or female sex in O. hubbsi. For this purpose I have done genotyping as well as histological studies. Finally, I have applied CRISPR/Cas9 gene knock-out system for the first time by myself with the help of my supervisor. This helped me a lot to understand the method and its applications.
Apart from the experiments, lab members and the administrative office was always very helpful and friendly to me. The very first week, NINS Office set a meeting for foreigners to inform us about the life in Japan and official works needed to be done by us. I found this meeting very informative and helpful. In this meeting, I have also met international students from different labs and we became very good friends. Later, we had chance to hang out all together and discover Okazaki. In my opinion, there might be even more meetings for foreign students to help them find friends during their stay. During my stay, several conferences, parties, and events took place and I have found this very helpful to get to know Japanese culture. Thanks to the localization of Okazaki in Japan, I travelled to Tokyo and Osaka to learn about the Japanese history, people, culture, and especially cuisine.
Overall, my lab work helped me to learn a new concept, apply new methods, and gain self-confidence, and more. I wish, I could even stay longer if I did not have to turn back to Germany for my thesis project. I need to thank to Prof. Dr. Kiyoshi Naruse, Dr. Yusuke Takehana, Dr. Saori Yokoi, Ms. Akiko Nishimura, and all other technicians and responsible people of the program. I am very grateful to them for giving me this opportunity to be in Japan. I hope, I can come back to Japan in the future, as well.
Mahidol University, Thailand
It was the first time that I visited Japan and NIBB. I am very grateful to attend NIBB internship program. This program extended my vision and gave me many new experiences.
I joined Dr. Kawade Kensuke’s laboratory which focused on plant development and physiology. This laboratory has been studying metabolic systems that coordinate with developmental progression in plants, especially in Arabidopsis. We are interested in ANGUSTIFOLIA3 (AN3) which is a signaling molecule involved in the control of proliferation of epidermis and mesophyll in leaves.
I had an opportunity to do some experiments including root growth kinetic comparison between AN3 wild type and an3 mutant, expression pattern of AN3 encoded GFP and AN3 promoter-ß-glucuronidase (GUS) reporter line in order to prove and illustrate expression of AN3 in root. The results from these experiments were consistent with the previous studies. I also had a chance to do root confocal analysis, leaf area and cell size experiment.
The Arabidopsis plants used in these experiments were grown in our laboratory. Root length and expression of AN3-GUS and AN3-GFP was observed by using confocal microscopy. The photos were captured in order to measure cell size by using Image J program. The area of Arabidopsis leaves were also measured by using Image J program and leaf palisade mesophyll cell number was calculated from cell size and leaf area.
This internship program is very good for students who are interested in Biology Science and would like to enjoy Japanese culture including working style, life style and so on. There are many new experiences you can receive here. Especially, for students who would like to be a master or Ph.D. student in Japan, This internship would be a great choice.
Okayama University Office, Viet Nam
My name is Nguyen Thi Hong Dung and I come from Vietnam. Currently, I am first year student of Okayama-Hue International Master’s program in sustainability of rural and Environmental systems. I was very happy when I was selected for NIBB internship program 2015. I was intern student at laboratory of Molecular Genetics for Reproduction from 5th November 2015 to 5th December 2015. During this internship, I focused two main experiments.
I had great internship. In one month, I studied a lot of skills and knowledge. Surely, this program was a rare chance for anyone who wishes to experience the professional study and research environment at NIBB. Through the internship program, with the guidance from professors and lab mates, I practiced research skills, including: Techniques, experimental operation, data recording and analysis, methods of data reporting, etc. Besides specialized knowledge and skills in scientific research, I learned the work ethic of scientists in one of the fore-running scientific labs of Japan – one of the leading countries in terms of discipline, enthusiasm, and hard work. During my internship, my lab mates not only helped me in my research but also in learning Japanese. They were very friendly and kind. I would like to thank all laboratory members: Naomi, Watakabe, Sakae, Furimoji, Nishimura, Kinoshita, and Otake. I would like to thanks Ms.Mariko Kikuchi, she was very thoughtful and kind, she helped me a lot in research work as well as daily living. With her helping, I was always very happy when I stayed in Okazaki city. Especially, Dr.Tanaka was very kind with me, he gave me a feeling of comfort. I will keep these great memories in my heart.
These experience would be very valuable to me, will help me adapt to and successfully complete my one year of master’s program at Okayama University as well as leading me to become good researcher in the future. I would like to thank Dr. Tanaka Minoru and NIBB for granting me this opportunity
Hacettepe University, Turkey
I have been working on photovoltaics for all my career. I applied to NIBB ınternship program with the aim of gaining an understanding in photosynthesis machinery. My project in Professor Minagawa lab included fluorescence measurements of Chlamydomonas Reinhardtii mutants in order to understand the relationship between different photo-protection mechanisms.
As a physics engineer, working in a biology laboratory environment was an amazing experience for me. I felt really comfortable in this international environment, with the kind assistance of all students, academicians and support office as well. I had the opportunity to discuss my scientific point of view with everyone, attend to weekly meetings and also an international conference. The atmosphere in the whole institute was very open and supportive, and besides the scientific point of view, there were several social events where I could experience the hospitality and friendship of people while enjoying Japanese culture and cuisine.
The accommodation at the Mishima Lodge was really comfortable with a good stuff. Everything was clean, safe and well organised. The lodge was really close to Institute, and also to the city center.
My time in this research internship was really efficient in scientific and also social point of view. Besides learning new methods and getting acquainted with the point of view of world class biology scientists, I had the opportunity to enjoy the well protected nature. Ultimately the time I spend in Okazaki was rewarding and valuable for me.
Universidad Nacional del Santa, Nuevo Chimbote, Peru
My name is José Alexander Carranza Luna from the Universidad Nacional del Santa, Nuevo Chimbote – Peru. I am graduated from the school of Biology in Aquaculture of the Faculty of Science.
I was an intern in the Laboratory of Molecular Genetics for Reproduction, which is led by PhD. Tanaka, Minoru from 4th December 2015 to 1st March 2016. I came to the NIBB after obtain a scholarship from the Peruvian government, specifically for the National and International mobilization program in science, technology and innovation from the Consejo Nacional de Ciencia, Tecnología e Innovación FONDECYT-CONCYTEC (Peru), in order to do a: “Training in techniques for sex determination and differentiation in fish” and contribute to the project: Sex Determination and differentiation in Arapaima gigas “Paiche” (the second freshwater fish largest in the world), which is executing in the Laboratory of Genetics, Physiology and Reproduction in Peru, which I take part.
During my stay at the NIBB I could learn various techniques such as: in situ hybridization, immunohistochemistry, confocal microscopy, genome editing by CRISPRcas9, probes designing, bioinformatics software and others, by using the Japanese fish Oryzias latipes “medaka”. I studied about the germ cells biology, how the gonad is form and what genes are important by analyzing several mutants strain. Since the germ cells are a set of very important cells because they are responsible to transmit the genetic information to the next generation and also have the ability to form a whole individual, I am very interesting for continue doing research in this fascinating field.
I would like to thank PhD. Tanaka, who gave me the opportunity to join the lab and for his suggestion in the experimental process in the Paiche project in Peru. Likewise, I would like to thank to the lab members: Shiraishi, Sakae, Fujimori, Kikuchi, Otake, Watakabe, Suzuki and Kinoshita for helped me with the experiments, for their kindness and their friendship. Specially, I would like to thanks to Nishimura-san, who was my mentor along this 3 months, thanks for his time, dedication, for the knowledge that shared with me every time, for introduce me to the Japanese culture and for his sincere friendship. Also, I would like to thank to the members of Miyanari's Lab who were friendly and kind.
Along this 3 months, I could visit some touristic places as Okazaki Castle, Takeshima Shrine, etc., which helped me to know and understand a more details the Japanese culture. Also, I could taste several typical food (Yakiniku, was one of the best) and enjoy incredible moments in this beautiful and fascinating country.
Eternally grateful.
VNU University of Science, Viet Nam
I continued my journey of science by applying to NIBB internship program, immediately after completing bachelor’s degree. As a newbie in this field, I expected this program would be opportunity to raise characters of scientist inside me: If I faced a problem, how could I handle it? How to solve a scientific question by designing plan or experiment?
Then I chose laboratory of Assoc. Prof. Kamei, because of his using medaka fish model and heat-shock system, as my undergraduate laboratory. Besides, I wanted to have a chance at IR-LEGO, the technique that using laser to induce expression of heat shock promoter-driving gene in desired cells. Nearly 3 months working in Kamei-lab, I was enthusiastically supported by members of this laboratory and also Bioresource Laboratory. Thank to that help, my experiments (evaluation of heat-shock response and molecular experiments) could turned fluently and be finished with some expected results. I think the obtained results were not significant to me than what I have learned: techniques, experiment manipulation and even working habit. And my questions at beginning also were answered somewhat.
To say something about this program, in unscientific aspect, I feel how lucky I was to join this program: coming to my dreaming country at the first time going abroad, being helped by surrounding people, immersing well in Japanese culture. Thank you for giving me precious experience and memories.
Heidelberg University, Germany
I am a developmental biology master’s student from Heidelberg University in Germany and I had the wonderful opportunity to join the lab of Assoc. Prof. Minoru Tanaka on Molecular Genetics for Reproduction. During my internship I focused on two topics, analyzing mutant lines and establishing a subcutaneous transplantation system in Medaka.
One female medaka mutant shows a defect in follicle formation, and some germ cells express the spermatocyte marker Shippo1. in situ hybridization turned up positive for Shippo1 for the mutant line, indicating spermatogenesis is occurring in the mutant ovary. Interestingly, ovaries from one mutant line were also able to fertilize WT ovaries. There is an implication that oocytes might regulate type I germ cell proliferation if the mutants show less oocytes and more type I germ cells. Therefore this was tested through BrdU incorporation of hatchlings and subsequent immunohistochemistry and type I germ cell counting. Unfortunately homozygous mutants were not obtained so this theory could not be tested.
For the other experiment, the purpose was to establish a subcutaneous transplantation system using olvas-eGFP/sox9b-DsRed donor gonadal cells in WT and host and observation of the donor cells after transplantation: how long do the donor cells remain in the host before rejection? As opposed to Zebrafish, using PBS in the water to sustain the transplanted fish does not work well for Medaka. We figured out the optimum conditions to keep the fish alive as long as possible; RO water with streptomycin/ampicillin that is changed every other day and an air bubbler in each tank. Retention of the donor cells depended on the transplantation technique so it was variable, however after the first week most of the GFP signal was gone and the DsRed was retained for longer while also decreasing.
After a lot of direct sequencing and three months later, I can safely say that coming to NIBB and Japan has been one of the most defining experiences of my life. Other than the new techniques I learned in the lab and exciting research I participated in, I was made to feel very welcome by everyone and there wasn’t anything I needed or was lacking. On the weekends I was able to experience what Japan has to offer from beautiful landscapes, mouth-watering foods and fascinating culture. I highly recommend this internship program.
Shanghai Jiao Tong University School of medicine, China
I am a medical student in Shanghai Jiao Tong University School of Medicine, China. I was an intern student at Professor Minoru Tanaka’s lab at The Department of Basic Biology, Laboratory of Molecular Genetics for Reproduction from 4th of August 2014 to 15th of August 2014. The main area of study was about Gonadal sex differentiation in Medaka by observing Germ cells.
Germ cells are a type of biological cells that involves in reproduction. In many animals; the germ cells originate in the primitive streak (structure that forms in the blastula during the embryonic development) and migrate via the gut of an embryo to the developing gonads. There, they undergo cell division of two types, mitosis and meiosis, followed by cellular differentiation into mature gametes, either eggs or sperm.
Germ cells play an essential role in Sex differentiation in Medaka. It is required for ovarian formation and also Germ cell deficient gonads develop a testis-like structure.
In vertebrates there are two modes for the determination of sex
1. Genotypic Sex Determination (GSD)
2. Environmental Sex determination (ESD)
• Mammals and Birds follow GSD.
In medaka sex-determining gene dmY/dmrt1y has been identified. My studies were mainly focused on GSD, two important genes.
At the beginning of the experiment I had to dissect Medaka lava to extract gonads which was a very interesting activity. I enjoyed collecting fish eggs at Myodaiji. Performing micro surgery was where I used my skills as a medical student who had assisted surgery at hospital. It was an opportunity to exchange knowledge with the PhD students at the lab. Therefore, my overall experience was invaluable. I certainly had a very useful time with Professor Tanaka and his team.
Bangalore University, India
My Name is Shashank Kumar from New Delhi, India. I have completed my Masters from Bangalore University in Biotechnology, I am very happy by the selection in NIBB internship program 2014 at National Institute for Basic Biology, Okazaki, Japan. I feel very lucky myself to have this opportunity and working in a high standard research laboratory. I was assigned at Division of Molecular and development biology under the supervision of Sir Prof. Shinji Takada. I sincerely thanks to Sir Prof. Shinji Takada for giving me this opportunity to work in his laboratory. I sincerely want to represent my heartily gratitude to all of the lab members who support me in every way.
I successfully completed the project and learned many advance techniques, which are going to be very helpful for my future research work. I am sincerely thankful to Mii San who has taught me different techniques, which are used in recent molecular and developmental biological research.
My primary objective was to experience basic DNA construction, which is obtained by double digestion of two restriction enzymes. The DNA were isolated and purified by gel purification method. We have used two specific type of strain (mutated and non-mutated). After the digestion we have done the gel purification and isolated the specific gene. This specific gene was then transformed to the competent cells of E.coli strain. After the incubation period the transformed E.Coli cells were observed and colonies were picked for mini culture, for the specific transformation we have done the colony PCR that indicate the successful transformation of the cell. All samples were successfully transformed and the result was obtained successfully. Apart from this Mii San has taught me about the Confocal Microscopy and Microinjection of XWnt mRNA in Xenopus laevis.
As a Biotechnology background, these experiments were totally new for me but I sincerely thanks to Mii San for guiding me in each and every step. He explained me all the basic principle for all the experiments and taught me the minor specification in the experiments, which is very useful for my future research. Even I am sincerely thankful to Sir Prof. Shinji Takada, Mii San and all the lab members, which has helped me a lot for my presentation. During my stay, Sir Prof. Shinji Takada and all lab members has given me a very warm welcome dinner party at an Indian Restaurant for that I am really thankful to them. As it is known to the world Japan is very rich in culture, I had a dream before to visit Japan and the dream came true due to this internship program. Even my all lab members were very keen to know about the Indian culture and I have told them about the Indian culture.
At last I would like to thank to Ritsue Takahashi sir, for guiding me from India to Japan. I would also like to sincere thanks to Ukai San for the administrative work and helping me a lot during the stay in Japan. I would like to sincere thanks to NIBB and Sir Prof. Shinji Takada for granting me this opportunity. I would also like to thank my all lab members for caring me a lot. I am also delighted for spending my whole tenure at Mishima Lodge; the facilities were very comfortable for making my stay delightful.
This Internship was very important for me to become a good researcher and a good human being for my future. I sincerely admire this country with most honest, hardworking and helpful people around there. I really want to continue my further research work and I hope I will get the chance again to carry out my research in this lab. Thank you very much.
University of Pécs, Hungary
My name is Gergo Palfalvi and I am from Hungary. I am a graduate biology student and my topic is the development of the pitcher shaped leaf of the Australian pitcher plant Cephalotus follicularis. This plant is a carnivorous plant that has a special leaf of pitcher shape dedicated to carnivore syndrome but also has a simple, non-carnivorous flat leaf. To decide which one is grown, the plant screens its environment. Kenji Fukushima from Prof. Hasebe’s laboratory found the temperature the main factor in this change. In January I visited Prof. Hasebe’s laboratory and I found that on cold temperature the nutrient deprivation cause higher pitcher development rate. This founding is the basis of my master dissertation’s topic.
In this time the reason of my visit was to investigate some in silico data from C. follicularis’s genome which is connected to the nutrient acquisition and to the carnivore syndrome, too. For this, first I learned the basic use of UNIX and shell scripts on NIBB’s super computer. After that I learned how to make a phylogenetic tree for homologous genes starting with BLAST search and finishing with the gene tree. I examined the plant’s messenger RNA (mRNA) transcriptome data and found differentially expressed nutrient transporters which can be good candidates for genes involved in carnivore syndrome. Now I can use advanced bioinformatic tools and I can start my own project at Hungary with microRNA (miRNA) transcriptomics which is highly connected to the project of Professor Hasebe and Mr. Fukushima.
There was a second aspect to my visit this time. In December I will take the examination for SOKENDAI and I would like to investigate the pitcher development in the future. To start this project I optimized in situ hybridization protocol for C. follicularis including tissue fixation, embedding, serial section making and in situ hybridization. This time I have succeeded with Histone H4 gene which is a good indicator for actively dividing cells. Now I could describe some part of the developing pitcher’s division areas but my data is not enough for conclusions yet. To describe the development phases of the pitcher leaf, I made serial sections from shoot apex grown on pitcher inducing- and also on flat leaf inducing environments and stained them with Toluidine blue. Now I am processing the data, but I can say it looks like the determination of leaf type occurs in a very early developmental stage of leaf primordia – which is opposite with previous thoughts and the development is quiet different from other non-related carnivorous plant’s pitcher leaves, such as Sarracenia purpurea (Mr. Fukushima’s project was the development of the pitcher leaf of S. purpurea).
On the extracurricular programs, I am wondering about the Japanese old and rich culture and it was a great opportunity to get to know with it. The atmosphere in the laboratory was always great and I have had opportunity to discuss with and get inspired by magnificent researchers.
After this trip I can say I am richer with a lot of knowledge, inspiration and great friends, in addition to a lot of experience about international collaborations.
I would like to thank you this opportunity and in the future I would like to join Professor Hasebe’s team to examine the evolutionary development of the pitcher leaf of C. follicularis.
University of Heidelberg, Germany
My name is Julian Stolper and I am a Masters student at the University of Heidelberg, focusing on developmental biology.
During my 3 month stay at NIBB in Prof. Naruses lab, my project consisted of establishing an enzyme linked immunosorbent assay (ELISA) in order to quantify insulin levels in Medaka (Oryzias latipes). Under the guidance of Dr. Shin-ichi Chisada, we managed to measure insulin levels of LepRKO mutants and wildtype Medaka under different feeding conditions.
Insulin levels in blood serum were quantified by performing competition experiments with known concentrations of biotinylated Medaka insulin (fractioned by high performance liquid chromatography). By using specific antibody combinations and horseradish peroxidase (HRP) reactions, the experiments could be analysed.
Working in Prof. Naruses lab was a great opportunity to broaden my horizons not only on a scientific level but also on a cultural level. I experienced great hospitality and a huge effort to bring me closer to Japanese culture, language and cuisine. Several trips during the weekend to different places in Japan with my Co-workers made my stay in Okazaki unforgettable. Acknowledgements go not only to Prof. Kiyoshi Naruse for letting me stay in his lab for 3 months but also to my coworkers and friends Dr. Shin-ichi Chisada, Dr. Yusuke Takehana and Ikuyo Hara and all the staff. Private tea ceremonies and lab get-togethers were very special moments for me in which I could experience great friendliness and interest also in German culture and language.
The NIBB internship program is a very well organized program which not only covered my flight and accommodation expenses but also made it easier to start my internship in Japan by always being there in case any question would arise. For this, I am deeply grateful.
In summary I can say that going to Japan was one of my best decisions so far and I am very honored and thankful for this opportunity which NIBB and its internship program provided me.
National Institute of Science Education and Research (NISER), India
am KhushbuKumari from Jamshedpur ,India.I have completed my Integrated masters in Life Sciences, major in Plant Biology from National Institute Of Science Education and Research,Bhubaneswar. I am very fortunate that I got selected for the NIBB Internship Program in the period from October 7th to November 7th,2014. I got the opportunity to work in Dr.Jun Minagawa lab, where I worked to study the correlation between state transition and Photoinhibition mechanism using state transition an quenching mutant strains of Chylamodomonas reinhardtii.
My internship helped me to gain lot of experiences and skill in Photobiology field as well as being able to communicate with outside culture and society. I am really greatful to Dr.Tokusu Ruytaro and Dr.Shun Takashsi who helped me in every aspect and explained me every query very patiently. During the course period I learned how to grow the Chylamodomona sreinhardtii cells in sterile culture system maintaining the optimized required conditions. I performed the experiments which includes SDS Gradient PAGE,Immunoblotting,Pluse amplitude Modulation Spectroscopy for measuring the Fv/Fmratio. My research basically focuses on to compare the Fv/Fm ratio between wild type,state transition and quenching mutants when exposed to high light illumination.
Right from the selection to arrival in Japan I was guided very carefully by Ms.RituseTakashai .Everyone in the lab was very friendly and of helping nature. I also got the opportunity to attend the Journal club every week where the recent published work of Photobiology and progress report was disused which helped me to broadened my ideas and a better understanding in this field. A special thanks to Yousef yari kamrani,Tirupati, Kishimoto mariko,Kosuge,Hirokato and other lab member who took very good care of me and made me feel more comfortable. My accommodation atMishima lodge was very close to the NIBB campus and easy accessible. During my stay of one month, I visited Nagoya and nearby places in Okazaki like Okazaki castle etc.I enjoyed Japanese culture a lot.
Finally my sincere thanks to NIBB for giving me this wonderful opportunity to develop my scientific skills and experience Japanese culture. Iwish all the best and great success to all the lab members and take this opportunity to thank them for their kind cooperation, hospitality and the great atmosphere. I hope I have another chance to visit NIBB again. Thank you .