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Comments on Johnathan W. Edmonds et al. (2007) International Worm Meeting "RNAi screen for oocyte genes that control directional sperm motility." (0)
Overview
Johnathan W. Edmonds, & Michael A. Miller (2007). RNAi screen for oocyte genes that control directional sperm motility presented in International Worm Meeting. Unpublished information; cite only with author permission.
The mechanism(s) by which motile sperm find oocytes in the female reproductive tract is not understood. To investigate this mechanism, we have developed an in vivo assay for tracking the movement of sperm in the uterus of C. elegans hermaphrodites. Previous results support the hypothesis that oocytes generate sperm-recruiting signals derived from polyunsaturated fatty acids (PUFAs). These signals direct sperm migration to the spermatheca, the site of fertilization. In mammals, PUFAs are synthesized into eicosanoids, which direct T-cell migration to sites of inflammation. I am using RNA-mediated interference (RNAi) to identify genes that function in oocytes to control directional sperm motility. Genome-wide DNA microarray (Reinke et al. 2004) and in situ hybridization (Kohara 2001) databases were screened for genes expressed in oocytes. I have identified eight genes that are required in hermaphrodites for the directional movement of wild-type sperm. Several of these genes are related to human genes implicated in eicosanoid synthesis and transport. I am currently focusing on the genes T28F3.1 and C01F6.1, which encode proteins called copines. Copines are conserved calcium-dependent membrane binding proteins with unknown functions. They are characterized by having two Ca(2+)-binding C2-domains at the N-terminal region and an A-domain at the C-terminal region. We hypothesize that C01F6.1 and T28F3.1 function in oocytes to target PUFA-modifying enzymes to the plasma membrane where they can generate sperm-recruiting signals. I am currently conducting phenotypic analysis of loss of function mutations in C01F6.1 and T28F3.1. Results from this analysis and site of action studies will be presented.