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.The long-term goal of the proposed studies is to elucidate the molecular events underlying the regulation of sperm capacitation and fertilization. Such knowledge is essential for developing therapeutic strategies to prevent and treat infertility and dysfunctions of sperm. In mammals, fertilization is triggered by the binding of an egg-derived glycoprotein, ZP3, to a specific receptor (ZP3-R) on the sperm plasma membrane. The resulting crosslinking of the ZP3-R activates a group of signaling proteins (gamete recognition molecules or GREM) that induce a variety of signaling events including increase in sperm hypermotility, binding of the zona pellucida, and acrosome reaction. The molecular components of the ZP3-R-mediated signaling pathway have been largely unknown. Recently, we identified two small peptides, histrelin and histrelin2, which mimic ZP3 in a receptor binding assay. These peptides bind to ZP3-R with high affinity and exhibit similar or higher affinity than ZP3. Interestingly, both histrelin and histrelin2 are present in the mammalian ovary, and can be detected on the ovulated oocyte and/or cumulus cells. The specific hypothesis to be tested in this project is that histrelin and histrelin2 interact with the ZP3-R, thereby initiating the ZP3-R-mediated signaling events. It is hypothesized that histrelin and histrelin2 bind to the ZP3-R and act as priming ligands that induce a conformational change in the ZP3-R, leading to the activation of signaling molecules. This hypothesis will be tested in the following two specific aims: 1) The function of histrelin and histrelin2 in ZP3-R signaling and fertilization will be examined using zona-intact hamster oocytes; 2) The functional domains of ZP3-R will be identified by a novel homology-based cloning strategy and site-directed mut be359ba680
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