Running Western Blots, Better Mummichog Cladograms

We're finally running the first test Western blot on brain tissues from scarlet and telescope shiners from the early winter. We're trying to measure the amount of an important subunit of the cellular membrane protein NMDA, a key channel for learning (and sex) for vertebrates like fishes and ourselves. This first run probably won't work too well since the lab group doing it haven't used this exact protocol. But on Saturday they hope to spend the whole day running a better calibrated Western so we can get reliable data. The fish in this experiment include young-of-the-year individuals, and I'm not sure what to expect with their NMDA levels. With any luck this will be the beginning of a lot of work with Westerns. We're going out to Limestone Creek tomorrow for a seasonal collection of scarlet shiners, in part for some NMDA tests. I hope to be able to wade without waders since the temperature is now in the 70's F for several days.

I met with Kris yesterday, and his cladograms of mummichog DNA are even better than before. For creating a neighbor-joining tree he ran 10,000 bootstrap replications, which indicates the confidence you can place in a tree. Several of our nodes are supported at the 100% level (all 10,000 replications showed these nodes), and the weakest one (separating the Charleston & Sapelo Island, GA, fish from the Virginia Beach & Chincoteague, VA, fish) is 63%. It's not really surprising, but I feel even better about what we've found and its interpretation (a strong founder effect for the post-glacial recolonization of the northeast as the ice retreated starting about 14,000 years ago and climate moderated).