Saturday, July 19, 2008

The program is ending is about two weeks. I'm sad because I definitely wish I had more time for research, but that's the way it is. I would be troubleshooting quite a few things, but the next weeks will be quite busy enough because I'll be getting most of my data then. After I get the data, I'll need to make the usual poster and Powerpoint presentation that goes along with such summer programs. I'm not nerve-wracked about presenting yet, but I might be when it comes to the actual day. First, I need those results! I recently got three different lines of flies(it took some time to create the mutant lines with the required markers, although I wasn't the one making them), which all have lacZ labelling the product of puckered (puc), which gives a measure of the Jun kinase (JNK) activity. One line is a wild-type control, and the other two are different alleles of raw, a mutant that affects larval dendrite morphology and the JNK pathway, in addition to some other affects. The raw mutants are homozygous lethal and the embryos die early, so these lines also have GFP, so I can label the heterozygotes and identify both genotypes. I need to stain these lines with certain antibodies labeling the neurons we are interested in, puc, and GFP to label the heterozygotes. I'll be spending some time trying to get good images of the patterns of expression in each line. The microscope for this takes some getting used to, and I'm not there yet. I did get to have a quick look at the control and mutant with my supervisor doing the imaging. It's pretty exciting. Raw mutants seem to have upregulated expression of puc. There is normally just one specific neuron that faintly expresses puc. The mutant is really messed up because raw also affects the development of the cuticle. The epidermis needs to close dorsally for the cuticle to exude from there, but in raw mutants it never does. It's a slow process to get these pictures because there's an ideal range of ages and orientations that I'm looking for, so it's good that I'm relatively patient. Wish me luck in finding some good embryos :)

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