NOVEL CONTRACEPTIVES: CONTROL OF OOCYTE MATURATION

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 该项目的目的是探索在灵长类卵巢中存在减数分裂激活和抑制基因的选择性表达的假设，以及针对这些基因将导致基于配子的非同源避孕药的开发。其具体目标是：(1)评估与灵长类卵巢恢复减数分裂有关的基因的表达，并调查选定的候选者的体外功能；(2)确定选定的药物是否可以在不改变猕猴卵巢其他功能的情况下扰乱卵母细胞的适时成熟；以及(3)确定选定的候选者是否可以在群体交配情况下作为避孕剂定期循环猕猴。在对INSL3-RXFP2信号通路在猕猴卵巢中的表达和功能进行深入分析后，我们得出结论：该配体-受体对不是猕猴卵母细胞成熟所必需的。因此，我们专注于一种新发现的卵母细胞特异性减数分裂抑制物，即WEE2，它以前在青蛙和小鼠中进行了研究。WEE2主要在猕猴的卵母细胞和着床前早期胚胎中表达，在睾丸中仅有微弱表达。当卵母细胞内cAMP含量较高时，WEE2的表达下调会导致泄漏减数分裂的恢复，而WEE2的过度表达则会延迟卵母细胞的自发成熟。我们目前正在研究分子中的关键功能区域，希望开发出抑制或模拟WEE2活性的化合物。此外，我们还在猕猴卵母细胞中对WEE2的中和酶CDC25进行功能研究。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The purpose of this project is to explore the hypotheses that selective expression of meiosis activating and inhibiting genes exists in the primate ovary, and that targeting these genes will lead to the development of gamete-based, nonhomornal contraceptive agents. The Specific Aims are: (1) to evaluate the expression of genes involved in the resumption of meiosis in the primate ovary and to investigate the in vitro functions of selected candidates; (2) to determine if selected agents can disrupt timely oocyte maturation without altering other ovarian functions in rhesus macaques; and (3) to determine whether selected candidates can function as contraceptive agents in regularly cycling rhesus monkeys in group-mating situations. After a thorough analysis on the expression and function of INSL3-RXFP2 signaling pathway in the macaque ovary, we concluded that the ligand-receptor pair is not essential for oocyte maturation in rhesus macaques. Thus, we focused on a newly identified oocyte-specific meiotic inhibitor, namely WEE2 that is previously studied in the frog and mouse. WEE2 is predominantly expressed in the oocyte and early preimplantation embryos in rhesus monkeys, and only weakly detectable in the testis. While down-regulation of WEE2 caused leaky meiosis resumption in the presence of high intra-oocyte cAMP, overexpression of WEE2 delayed spontaneous oocyte maturation. We are currently investigating key functional domains in the molecule, in hope of developing compounds that inhibit or mimic the activity of WEE2. In addition, we are also conducting functional studies on a counteracting enzyme of WEE2, CDC25, in the macaque oocytes.