NOVEL CONTRACEPTIVES: CONTROL OF OOCYTE MATURATION

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 本项目的目的是探索灵长类动物卵巢中存在减数分裂激活和抑制基因的选择性表达，以及靶向这些基因将导致开发基于配子的非同源避孕药的假设。 本研究的具体目的是：（1）检测灵长类卵巢减数分裂恢复相关基因的表达，并探讨其与卵巢成熟的关系。 所选候选物的体外功能;（2）确定所选试剂是否可以破坏恒河猴中卵母细胞的适时成熟而不改变其他卵巢功能;和（3）确定所选候选物是否可以作为 避孕药在定期循环恒河猴群交配的情况下。 通过对INSL 3-RXFP 2信号通路在猕猴卵巢中的表达和功能的深入分析，我们得出结论：配体-受体对不是猕猴卵母细胞成熟所必需的。 因此，我们专注于一个新发现的卵母细胞特异性减数分裂抑制剂，即WEE 2，这是以前在青蛙和小鼠的研究。 WEE 2主要在恒河猴的卵母细胞和早期植入前胚胎中表达，仅在睾丸中微弱检测到。 虽然WEE 2的下调导致在高的卵母细胞内cAMP存在下的减数分裂渗漏恢复，但WEE 2的过表达延迟了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结构域，希望开发抑制或模拟WEE 2活性的化合物。 此外，我们还在猕猴卵母细胞中对WEE 2的抵消酶CDC 25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. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.