The Williams lab undergrads are BACK and ready for a whirlwind of a semester. Our fearless lab-tech, Anna Marie, no longer has to hold down the fort alone because Ken Brit, Marcus, and I are officially ONE week into our last semester at Bates. For me, excitement about this final semester and my thesis is coupled with an underlying sense of panic about both life after Bates and somehow completing a thesis in the next 11 weeks. Full steam ahead!
I spent this past summer working with one of Larissa’s colleagues at the University of Massachusetts in Amherst. There I learned zebrafish and toxicology basics that are going to be incredibly useful for my transition into the Williams lab. At the same time, I am realizing that things at Bates are done a bit differently than at UMASS. I have some slightly different protocols and schedules to get used to.
While at UMASS I also began thesis data collection. I did a chronic (120 hr) MEHP exposure with wild type and mutant Nrf2a embryos over the summer. When the fish were about 15 days old, I stained their lipid depots with Oil Red O. Preliminary data indicated a relationship between increased lipid content in the liver and MEHP exposure. Furthermore, we found that this phenotype was exacerbated in the mutant line.
My task for the winter? Figure out WHY we saw this phenotype by looking at the gene targets from my summer samples. I will also repeat this experiment with a Nfe2 knockout line. Right now, I need to figure out what is UP with my cDNA from the summer. There have been some problems with the housekeeping genes. Our plan of action is to repeat qPCR with the cDNA stock and HOPEFULLY get our hands on the RNA from the summer (which is currently hiding somewhere in a freezer at UMASS).
Here’s a detailed list of what I’ve been up to this week:
• Toured the lab, picked a bench for the semester
• MET THE FISH
o I am a little embarrassed to say that I think I actually missed the fish from this past summer
o It is in my wildly unscientific opinion that the more you talk to your fish and give them love, the better their clutches will be and the easier your experiments will be. So. It is my goal to love these fish as much as I possibly can over the next 11 weeks.
• Animal Care Training
o We are officially allowed to hang out with the fish.
• READING READING READING everything I can about MEHP, ROS, Nfe2, hepatic steatosis… and the like
• Practiced pipetting/ qPCR with Anna Marie using one of the house-keeping genes that has been causing us trouble
• Ordering materials for my chronic exposure
o We needed some supplies for my chronic exposure experiment, including the lipid stain (Oil Red O) but also some little nit-picky things like an extremely specific size transfer pipette that I SWEAR by for the staining protocol.
• WRITING WRITING WRITING! So far, I have a few pages of both my methods and my introduction (oh boy oh baby! Here we go!!!)
• We ended the week with our first lab meeting. At lab meeting Larissa gave an overview of the investigative purpose of the lab and each of our projects.
The Nfe2 knockout fish are still pretty little, but they bred for this first-time this past week and it is my hope that I can set up my chronic exposure experiment on Wednesday!
Just keep swimming!