The research continues!
Now into the second half of our summer at Bates, we have all begun to fall into a standard routine in the Williams Lab.
We began this week with Michelle’s journal club presentation on the Liao et al. 1997 paper titled “The zebrafish gene cloche acts upstream of a flk-1 homologue to regulate endothelial cell differentiation”. Endothelial cells are the inner lining of many organ systems including the blood vessels. Overall, this paper was able to elucidate more of the signaling cascade that regulates differentiation during development.
This week I have been practicing the ChIP protocol again, however I have also been facing some difficulties with the protocol. In order to fragment the DNA so that in can be immunoprecipitated with an antibody I use a sonication machine that creates sound waves to break up the DNA. However, with the past two attempts the DNA has remained in its genomic form and not become fragmented. Hopefully, I will be able to sort this problem out and move forward with that portion of my work. I have also been designing qPCR primers for quantitative analysis of different XREs in the nuclear-related-factor (Nrf) gene family. I am excited for those to come in and see what data we will get from that project.
This week Michelle has continued to build her library of images from the confocal as well as create more composite images of the swim bladder and otoliths. It’s really amazing that she is able to visualize such small structures in the zebrafish embryos. Michelle has also begun learning how to microinject into zebrafish embryos. With this technique she can inject morpholino into the developing embryos and visualize what happens to the two structures in a “loss of function” assay. Nancy has been continuing to dose embryos and collect images for her work looking at the effects of phthalates like MEHP on gene expression. It’s amazing how fast the summer is going and we are all very excited to continue to delve into each of our projects.