UGR Blog Post 2

Fertilization is a complex molecular interaction that may require multiprotein complexes on both gametes for sperm-egg fusion to occur. My project aims to identify how potential multiprotein protein complexes are functioning and what their impact is on fertility and sperm-egg fusion. We have previously identified a sperm membrane protein interactome in Caenorhabditis elegans (C. elegans) that provided evidence that sperm membrane proteins interact extensively. My lab team has weekly meetings where we discuss our projects for the following week. I have been maintaining my worms and making sure they are healthy for future experiments. One of the difficulties I have faced is that one of my plates got infected, and this required me to follow better sterilization procedures.

We are analyzing the spe-36 gene, which was originally identified as a sterile strain. Further analysis revealed that spe-36 mutants are unable to fertilize eggs normally, but the cause is unknown. To better understand why spe-36 mutants are sterile, we are analyzing the mutants at different developmental stages. We are investigating three different strains N2 (wild-type), spe-36(as1), and spe-36(as1)asEx96. The spe-36(as1) mutant is the C. elegans without the gene of interest. And the spe-36(as1)asEx96 is the C. elegans with the gene of interest replaced and tagged with green fluorescent protein (GFP). We have conducted a brood size analysis at 16°C to assess fertility rate. We discovered that C.elegans with spe-36 mutants are unable to fertilize. When the gene is knocked in, the fertilization reaches close to the wild-type but not entirely. We are interested in seeing what will happen at 20°C and 25°C. We expect to see a slower fertilization rate at these temperatures since they are not the preferred temperature.

We have also identified two C. elegans strains with mutations in genes known to be necessary for fertilization to analyze the effect that missense mutations in multiple sperm membrane proteins have on sperm-egg fusion. Since I will be examining more than one mutation, we ordered different strains of C. elegans from the Caenorhabditis Genetics Center.The VC40852 strain has missense mutations in fer-1 and spe-10 and the VC20575 has missense mutations in fer-1 and spe-9. We are measuring the effect of these mutations on brood size and sperm-egg fusion. The analysis of both strains will provide insight into the genetic interactions between fer-1, spe-9, and spe-10 and allow us to generate a model of how these genes could be functioning together to mediate fertilization.

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