Research

Research Description

Osmoregulation and Contractile Vacuole Function Contractile vacuole complexes (CVCs) are widely distributed in protist and are also found in freshwater sponges and zoospores of some fungi. The CVCs are thought to be osmoregulatory organelles as their contraction frequency and the diameter of their contractile vacuoles (CVs) change in response to changes in the extracellular osmolarity. In last two decade, immunological and molecular biological and electrophysiological studies demonstrated abundant proton-translocating V-ATPase in its membrane that could provide the energy, from proton electrochemical gradients, for moving ions into the CV to be followed by water. The detailed contents of our progresses were well documented in following review article. Allen and Naitoh. 2002. Int. Rev. Cytol., 215, 351-394.
Selected Publications

Ishida, M., Aihara, M. S., Allen, R. D., Fok, A. K. 1993. Osmoregulation in Paramecium: the locus of fluid segregation in the contractilevacuole complex. J. Cell. Sci., 106, 693-702.
Fok, A. K., Aihara,M. S., Ishida, M., Nolta, K. V., Steck, T.L., Allen, R. D. 1995. The pegs on the decorated tubules of the contractile vacuole complex of Paramecium are proton pumps. J. Cell Sci., 108, 3163-3170.
Ishida, M., Fok, A. K., Aihara, M. S., Allen, R. D. 1996. Hyperosmotic stress leads to reversible dissociation of the proton pump-bearing tubules from the contractile vacuole complex in Paramecium. J. Cell Sci., 109, 229-237.
Naitoh, Y., Tominaga, T., Ishida, M., Fok, A.K., Aihara, M.S., Allen, R.D. 1997. How does the contractile vacuole of Paramecium multimicronucleatum expel fluid? Modeling the expulsion mechanism. J. Exp. Biol., 200(4), 713-721. w/cover picture
Fok, A.K., Yamauchi, K., Ishihara, A., Aihara, M.S., Ishida, M., Allen, R.D. 2002.　The vacuolar ATPase of Paramecium multimicronucleatum: gene structure of the B subunit and the dynamics of the V-ATPase-rich Osmoregulatory membranes. J. Euk. Microbiol., 49(3):185-196.
Ishida, M., Hori, M. Ooba, Y., Kinoshita, M., Matsutani, T., Naito, M., Hagimoto, T., Miyazaki, K., Uedaa, S., Miura ,K., and Tominaga, T. 2021. A functional aqp1 gene product localizes on the contractile vacuole complex in Paramecium multimicronucleatum. J.Euk. Microbiol., 68(1), e12843. doi:10.1111/jeu.12843 → Pre-Peer reviewed version of Ishida et al 2021 is available for download.

Intracellular Digestion and Membrane Trafficking Cellular membranes are made in a cell's biosynthetic pathway and are composed of similar biochemical constituents. Nevertheless, they become differentiated as membrane components are sorted into different membrane-limited compartments. Paramecium is a multi-polarized cell in the sense that several different pools of membrane-limited compartments are targeted for exocytosis at very specific sites at the cell surface. Paramecium has provided an excellent model for studying the complexities of membrane trafficking in one cell using both morphological and immunocytochemical techniques. This cell also promises to be a useful model for studying aspects of the molecular biology of membrane sorting, retrieval, transport, and fusion. The detailed contents of our progresses were well documented in following review article. Allen and Fok. 2000. Int. Rev. Cytol., 198, 277-318.
Selected Publications

Allen, R.D., Bala, N.P. Ali, R. F., Nishida, D.M., Aihara, M.S., Ishida, M., Fok, A. K. 1995. Rapid bulk replacement of acceptor membrane by donor membrane during phagosome to phagoacidosome transformation in Paramecium. J. Cell Sci., 108, 1263-1274.
Ishida, M., Aihara, M. S., Allen, R. D., Fok, A. K. 1997. Acidification of the young phagosomes of Paramecium is mediated by proton pumps derived from the acidosomes. Protoplasma, 196, 12-20.
Yamauchi, K., Aihara, M.S., Ishida, M., Allen, R.D. and Fok, A.K. 1999. Cloning and sequencing a protein involved in phagosomal membrane-associated protein in Paramecium. Mol. Biol. Cell, 10, 1031-1041.
Ishida, M., Allen, R. D., Fok, A. K. 2001. Phogosome formation in Paramecium: roles of cilia and solid particles. J. Euk. Microbiol., 48(6): 640-646.
Fok, A.K., Aihara, M.S., Ishida, M. and Allen, R.D. 2008. Calmodulin Localization and its effects on endocytic and phagocytic membrane trafficking in Paramecium multimicronucleatum. J. Euk. Microbiol. 55(6), 481-491.
Ishida, M., Aihara, M.S., Allen, R.D., Fok. A.K. 2011. The native structure of cytoplasmic dynein at work translocating vesicles in Paramecium. Eur. J. Cell Biol., 90, 81-92. w/cover picture

Basic Cell Biology of Paramecium By collaboration with other Labs, several basic cell biological issues, for example, nuclear dimorphism, endosymbiosis, ciliary movement, etc. have been worked on in our laboratory. In these issues, we were making mainly the morphological technique (immunocytochemical and ultrastructural) supports and participated in discussions. Furthermore, we keep having interests in introducing the new technology to the experimental system of a Paramecium.
Selected Publications

Ishida, M., Nakajima, Y., Kurokawa, K., Mikami, K. 1999. Nuclear behavior and differentiation in Paramecium caudatum, analyzed by immunofluorescence with anti-tubulin antibody. Zool. Sci. ,16, 915- 926.
Ishida, M., Kouda, A., Okumura, K., Nishiyama, N., Yamauchi, K. 2000. DNA fluorescence in situ hybridization in Paramecium: telomere localization in macronucleus. Zool. Sci., 17, 1289- 1295.
Nishiyama, N., Sawatsubashi, S., Ishida, M., Yamauchi, K. 2001. Organization and expression of the Paramecium caudatum gene encoding nucleosome assembly protein 1. Gene, 280(1-2), 107-114.
Nakajima, Y., Ishida, M., Mikami, K. 2002. Microtubules mediate germ-nuclear behavior after meiosis in conjugation of Paramecium caudatum. J. Euk. Microbiol., 49(1): 74-81.
Omura, G., Ishida, M., Arikawa, M., S. M. Mostafa Kamal Khan, Suetomo, Y., Kakuta, S., Yoshimura, C., Suzaki, T. 2004. A bacteria-free monoxenic culture of Paramecium bursaria: its growth characteristics and the reestablishment of symbiotic relationship with Chlorella in a bacteria-free condition. Jpn. J. Protozool., 37(2), 119-130.
Kutomi O, Hori M, Ishida M, Tominaga T, Kamachi H, Koll F, Cohen J, Yamada N, Noguchi M. 2012. Outer dynein arm light chain 1 is essential for controlling the ciliary response to cAMP in Paramecium tetraurelia. Eukaryot. Cell, 11, 645-653.
Kawano, M., Tominaga, T., Ishida, M. and Hori M. 2020. Roles of Adenylate cyclases in ciliary responses of Paramecium to mechanical stimulation. J. Euk. Microbiol., 67, 532-540. https://doi.org/10.1111/jeu.12800 w/cover picture
Hori, M., Tominaga, T., Ishida, M. and Kawano, M. 2023. RNA interference reveals the escape response mechanism of Paramecium to mechanical stimulation. Biophysics and Physicobiology. doi: 10.2142/biophysico.bppb-v20.0025.

Development of Teaching Materials for Science Education Our Lab has developed several teaching aids for science education, as a part of student’s graduation thesis. With consideration for use in the field of education, these teaching materials were published on HP (sorry for all in Japanese), in which we paid special attention for using rather simple way of expressions, user-friendly style of the format, and the colors. And also, in order to reply the question from children, teachers, or ordinary peoples the bulletin board was installed in these HPs.
Selected Publications

Takami, M. and Ishida, M. 2005. Pictorial Book for Planktons.
Aoki, A., Arai, I., Saito, H, and Ishida, M. 2007. Pictorial Book for Insect and Soil Organisms.
Nishino, M. and Ishida, M. 2012. Pictorial Book for Bugs Fond in Kindergarten.

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