The results of my research have been reviewed in detail in my previous posts, but there are two major outcomes of my work so far:

1. The ISHs that I completed this summer confirmed my previous results : perturbing Notch signaling dramatically effects expression of neural genes in early neural tube-tailbud stages, but these differences become less apparent in hatching stages and are not visible in swimming tadpole stages.

2. RNA extraction is difficult to optimize….

The second one is not as exciting as the first, but I have a feeling we’re almost there! Once we optimize the protocol it will be only a matter of days until I am able to have some qPCR results to quantify my qualitative ISH data. I am currently taking photos of my second round of ISH and also collecting expression data. I’ve also recently begun compiling all of my embryo injection data (rates of mortality and abnormal morphology). I am excited to bring these to the summer research showcase!

These results raise several interesting questions. The first is what molecular mechanisms are at work here that could be contributing to the restoration of a normal neuronal population? To answer this we will be injecting more embryos and assaying for genes involved at different points of neural differentiation. We also intend to use BrdU as well as TUNEL assays to stain for cell division and apoptosis, respectively. Finally, how can we test the effects of Notch on neurotransmitter phenotype specification? We have several ideas for how to approach this. The first is to inject constructs that are conjugated to a hormone receptor and would therefore be inducible. We also may try exposing cell cultures to pharmacological Notch agonists and antagonists and analyze gene expression. The exciting news is that thanks to this summer’s work I have many directions in which to go for my thesis!