After testing out some of my probes last week and determining that they are lethal at high concentrations, I took a different approach to injecting them in hopes of more informative data. First, I injected the probes at lower concentrations. This experiment produced more informative results; I found that although the oocytes still don’t go through anything resembling meiosis, they don’t explode (as often). Additionally, as expected, oocytes experienced high Rho activation throughout, almost as though I had unplugged something important that was inactivating Rho before… This period of high Rho activation made the cells turn completely white, since the Rho probe lit up all over the cortex. This led to the mutant's current nickname: 'whiteout'. As mentioned above, unfortunately, not all oocytes remained unharmed during the duration of my experiments. After addition of hormone to those oocytes injected with high concentrations of either my PBR or Snowflake-PBR probes, I found that the oocytes would try to contract as if they were trying to start meiosis, but would fail quite dramatically. A video is attached – warning, it’s a little gory! (sorry for the low quality)
Apart from accidentally exploding some oocytes this week, I have made good progress on other aspects of my project. My ‘dog’ (Rock regulatory domain) is almost ready for injection, with the ‘cat’ (Rock kinase domain) not far behind. Also, the pieces (front, middle, and tail) of the S. purpuratus Rock are ready for Gibson assembly: a cloning method that allows for assembly of multiple small fragments into a single cohesive sequence. This will allow us to finally construct a probe specific to an echinoderm Rock. Although I probably will not have a chance to include ‘cat’, ‘dog’, and the S. purpuratus data on my poster I will still try out these probes before I leave in hopes of confirming my preliminary results. Outside of lab, OIMB had its notorious ‘Spineless Soiree’ (otherwise known as the invertebrate ball) today. We all dressed up as our favorite marine invertebrates and strutted down a catwalk in the middle of our dining hall. I went as the purple urchin, S. purpuratus, along with Renee and Megan. We all carefully crafted urchin spines out of purple wrapping paper and huddled together with our spines in all directions. We also brought a little lantern to represent Aristotle's lantern. Although it was hot, messy, and pretty ridiculous, it was still a lot of fun!
0 Comments
Leave a Reply. |
AuthorI am a rising junior at Carleton College, majoring in biology with minors in Neuroscience and Russian. I'm very excited to be working in Dr. George von Dassow's lab this summer, where I will be studying cell biology and embryology of marine invertebrates. Archives
August 2019
Categories |
Proudly powered by Weebly