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

Anjali Mone, Biology (on right in photo)

Faculty mentor: Dr. Marcia O’Connell

              Our lab is interested in early embryogenesis in zebrafish. In particular we are investigating maternal factors because these are responsible for regulating early patterning events before embryonic genes are activated. In order to identify important regulatory factors, one approach that has been successful has been to identify products of genes that are important in invertebrates and find homologs of these genes in vertebrates. One maternal factor that is important in Drosophila is the product of squid gene.  Squid is responsible for forming the dorsal side of the embryo in Drosophila. Since squid has been found to be critical in early patterning events in flies, it is possible that a homolog of squid could play an important role in zebrafish development. We started by attempting to clone a homolog of squid. Subsequently, we began experiments to determine the function of the squid homologs in zebrafish.  Through RT-PCR we have found that this homolog is expressed in fish. Restriction digest was performed in which the gene was cut at a specific point in its sequence. We are also testing to see if squid protein is present in fish through western blots. Currently we are trying to assess the function of squid in zebrafish with morpholinos in which we essentially “knock out” a particular gene. In future experiments we would like to clone the squid homolog and sequence it. We will also determine if squid is strictly expressed during embryogenesis or if it is also present in the adult as well.

Personal Statement

              As I introduced myself to the summer research students for the third summer, I am asked the difficult question of why I keep coming back.  I thought back to my first summer in the summer undergraduate research program and how nervous I had been. Although I had some understanding of PCR and gel electrophoresis, I found that nothing in my genetics book would prepare me for the excitement that came with seeing bands on a gel. I found that as a learning student I was bound to have my fair share of broken beakers and hardened gels, but the few times that I was able to figure something out on my own made the experience worthwhile. What I really enjoyed about the research was not the science aspect of the research but the friends and contacts I made.  I thought back to my birthday and how they had surprised me with a trip to Philly.  I also remembered all of those trips to the beach, canoeing, berry picking, movie nights, barbeques, bowling and even those times we made dinner together.  After a long pause, I smiled and answered the question about what aspect of SURP kept my interest with the short but complete answer, I knew it would be another productive and more importantly, an enjoyable summer.