Summing it All Up. What an Amazing Summer!

In summary, my summer research project was highly successful in getting a head start on the Tweety research project. While full characterization of the Tweety genes’ expression is still in progress, the ten summer weeks that I spent on the project allowed me to get a thorough idea of the expression and perturbation patterns of the Tweety homolog 1 gene. In the final weeks of the summer, I began to use lacz staining with red-gal to determine the side that received the injection. This entails a similar process of injection to the GFP where I mixed red-gal mRNA with the Tweety mRNA injection mix. However, I no longer to need to side sort the embryos prior to fixing because re-gal staining is a nuclei stain that will last through MEMFA fixing and even an in situ experiment!

Overall, over expression of the Tweety homolog 1 gene through microinjection of mRNA has resulted in an irregular neural phenotype. This has been determined through in situ hybridizations of NBT, VLGUT, GAD, and Sox2. While NBT, VGLUT, and GAD are all downregulated on the mRNA injected side, Sox 2 is upregulated in the injected side of the embryo. Sox2 is typically in neural stem cells, thus suggesting that the Tweety injected side’s neural cells are not developing at the same rate as the uninjected side’s cells with more cells in a progenitive state.

For future research, it will be interesting to assess expression of PCNA, a proliferation marker, to see if there are any differences in cell proliferation between injected and uninjected sides of the embryo. In addition, throughout the semester, I will be focused on establishing a similar baseline of perturbation for Tweety 2 and Tweety 3 injected embryos. Already, Tweety 2 perturbed embryos appear to be reacting to the injection differently than the Tweety 1 embryos do. In addition, I will further explore the effects of interrupted expression of these three genes through morpholino experiments.

In conclusion, this very productive summer practically flew by as I immersed myself in my research and enjoyed every minute of it. I greatly look forward to continuing my research with Dr. Saha and her lab throughout the semester to come!