Investigating Homologs of the Squid Gene During Zebrafish (Danio rerio) Embryogenesis

Swapnil Patel, Biology (on left in photo)

Faculty Mentor: Dr. Marcia O’Connell

Our research is based on the gene, squid (sqd) found in fruit flies (Drosophila melanogaster). Innovative research conducted by Dr. Amanda Norvell and her colleagues has demonstrated the integral role of sqd and its resulting protein products on fly embryogenesis. The gene sqd is alternatively spliced yielding three different isoforms called SqdA, SqdB, and SqdS. Sqd is a heterogeneous nuclear RNA-binding protein, hnRNP, a class of proteins that bind to RNA to carry out their functions. Specifically, Sqd proteins bind to gurken (grk) mRNA and influences the localization and translation of it. The production of Grk protein, which acts as a ligand to the epidermal growth factor receptor (Egfr) of follicle cells surrounding the oocyte, stimulates the generation of a distinctly dorsal side. In the context of our research, we are investigating whether a gene similar to sqd exists in the zebrafish genome. We are currently investigating potential homologs, their sequence, function, in what tissues they are expressed and their role in vertebrate development.

This summer we examined two zebrafish genes that most closely matched sqd in nucleotide sequence. These gene sequences are called hnRNP A/B and Predicted Protein and we hypothesized that their proteins must function similarly to Sqd. To test our hypothesis and gather evidence to support it, we conducted a series of experiments. First, we extracted and purified mRNA and demonstrated through RT-PCR that both genes are expressed during embryogenesis. We also extracted proteins from the embryos and are currently examining the collected protein via Western Blots. In addition, we are investigating the function of the Predicted Protein and hnRNP A/B proteins via morpholino injection to prevent their synthesis in live embryos.

Personal Statement

              The Summer Undergraduate Research Program (SURP) has been a truly inspiring experience for me as a rising Sophomore Biology student at The College of New Jersey. Just stepping into the lab and becoming adjusted to its atmosphere is advantageous for those students who are likely to pursue graduate school. After obtaining unexpected results and attempting to troubleshoot, I learned how to approach a problem and using my knowledge, find the solution. Of course, the SURP experience is more than just the laboratory work, it is also an opportunity to meet other students who share my interests. The social aspect of the program not only makes the experience more enjoyable but also instills a sense of camaraderie among the participants especially with the relaxing weekly get-together events offering free food. So, for me SURP was intellectually stimulating, fun, and not to mention the personal gratification of contributing to a worthwhile endeavor where my efforts can make a difference.