Progress and Presentations

My PRMT-5 worms finally arrived! I also have a schedule for bleaching and synchronizing the worms and will be doing that shortly. Though I thought the hardest part would be the tests themselves it seems the C. elegans are the most troublesome. The delay in receiving them has at least given me a lot of time to work through my methodology and procedures.

C. Elegans culture work

In January, so far I've created more agar plates and poured them, inoculated the plates with E. coli and transferred the current worms I had onto new plates. Now, I'll be mixing the H2O2 (for inducing DNA Damage) and the bleach solution for the worms. I have the AO stain so as soon as I can synchronize and treat the worms with the H2O2 I can begin collecting data!

Semester 2 begins!

My worms are currently nice and alive and although I never found a source to irradiate them, I've decided instead to use DNA alkylating agents or other DNA-damage inducing agents like MNNG, MMS, NaHSO₃. Those are the three I'm currently considering. I hope to run my first tests soon and to get more comfortable with the culture of C. Elegans and the processes for AO staining and the DNA damage treatments. My goal for January is to run preliminary tests and to continue to learn about the culture of the worms. As soon as I receive the materials for DNA Damage I can start my first official test and begin to collect data.

Semester 1 Update

By the end of the semester my materials came and I've been able to start plates with the worms on them. This is super exciting, however, I still need to find somewhere I can irradiate the worms with the correct radiation. I'm looking to use a x-ray machine, and although I've sent emails and some follow ups, I haven't found a place I can go to yet. Since the tests I'm performing are relatively straightforward this is the last challenge I have before I can get started performing actual tests. Hopefully over the break I'll be able to find a source of radiation and be able to start when I return for second semester. Luckily, with all the time I've had, I've been able to write out detailed methods for my tests and processes so I feel very prepared to start working.

Refined Focus

Now, instead of attempting RNAi experiments and HIF-1 western blots, I plan to examine if the inactivation of prmt-5 (in apoptotic pathway) leads to excessive apoptosis in germline following IR treatments, which was found to likely be due to transcription of EGL-1. So, I'll compare PRMT-5 mutants (gk357) with Wild Type and measure germ line apoptosis after IR treatment through Acridine Orange staining. If apoptosis is increased, then I'll then compare PRMT-5 with PRMT-5 and EGL-1 double mutants with the same treatments. Hopefully, the materials I've been waiting on will come and I can start actually working after a while of waiting. I'm excited and sort of dreading seeing these little worms again.

My main challenge going forward will be finding a source of Ionizing Radiation. The studies I've read use 120 Gy, which is very high and pretty hard to find (and not something we have at MA either). Hopefully I'll be able to use an x-ray machine at least to irradiate the worms.

New Goals, Steps, and Project!

Well, I’m back, I’m a senior now, and summer is over. This summer, I thought a LOT about my MARC project and read maybe 30 journal articles (if I’m estimating on the lower end), and luckily, all this reading paid off as I found something I find interesting relating C. elegans, what I studied in the Spring and my other area of interest, cancer. In addition, I attended a two week biomedical research program where I got to do a bunch of labs and learn a lot about molecular biology at level I’d never learned before. I also got to perform a Western Blot lab, which is something I’ll likely need for my project this year so it was really great to successfully complete that. This September, I hope to start on my project and working in the lab, but before that, I’ll need to contact a mentor, either at the Buck Institute or at Dominican with someone who has experience working with C. elegans and can help me refine my focus and give me a starting point or a more streamlined place to start. So, I’ll be sending emails out and also looking into the materials necessary for the lab protocols I want to use.