NOVEL CONTRACEPTIVES: CONTROL OF OOCYTE MATURATION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 这笔赠款的目的是探索这样的假设，即灵长类卵巢中存在有选择性地表达减数分裂激活和抑制基因，以及候选分子可以通过建立的体外和体内实验序列来测试避孕潜力，最终在猕猴繁殖群体中进行避孕试验。其具体目标是：(1)鉴定可能与卵母细胞成熟相关的卵巢表达基因；并研究选定的候选基因(例如INSL3-LGR8途径、WEE1B和卵母细胞特异性的NLRPs)的体外功能；(2)确定选定的候选基因是否可以在体内用作恒河猴的避孕剂；以及(3)确定选定的候选基因(例如INSL3)是否可以在配对交配情况下作为避孕剂定期循环恒河猴。实验设计将包括：鉴定猕猴卵巢中的基因产物(mRNA和蛋白质)、卵母细胞中的体外RNAi以及未成熟卵母细胞和颗粒细胞与减数分裂抑制剂和激活剂的体外孵育(目标1)；在受控卵巢刺激(COS)和自然月经周期期间对猕猴体内注射INSL3或其拮抗剂，以及内分泌和毒性测量(目标2)；以及对繁殖群体中具有生育能力的雌性猕猴长期给予INSL3或其抑制剂以评估避孕效果和长期毒性(目标3)。这一方法有望为未来INSL3在人类身上的试验提供基础；并确定其他卵母细胞特有的新基因产品作为未来潜在的避孕目标。

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 grant is to explore the hypotheses that selective expression of meiosis activating and inhibiting genes exists in the primate ovary, and that candidate molecules can be tested for contraceptive potential using an established sequence of in vitro and in vivo experiments culminating in a contraceptive trial in breeding groups of macaques. The Specific Aims are: (1) to identify ovary-expressed genes that may be related to oocyte maturation; and to investigate the in vitro functions of selected candidates (e.g, INSL3-LGR8 pathway, WEE1B, and oocyte-specific NLRPs), (2) to determine if selected candidates can be used as contraceptive agents in rhesus monkeys in vivo, and (3) to determine whether selected candidates (e.g., INSL3) can function as a contraceptive agents in regularly cycling rhesus monkeys in paired mating situations. Experimental designs will include: characterization of gene products (mRNA and protein) in the macaque ovary, in vitro RNAi in the oocyte and in vitro incubation of immature oocytes and granulosa cells with meiotic inhibitors and activators (Aim 1); in vivo administration of INSL3 or its antagonist to monkeys during controlled ovarian stimulation (COS) and spontaneous menstrual cycles, plus endocrine and toxicity measurements (Aim 2); and chronic administration of INSL3 or its inhibitor to fertile females in a breeding colony to assess contraceptive efficacy and long term toxicity (Aim 3). This approach is expected to provide a foundation for future trials of INSL3 in humans; and to identify other oocyte-specific novel gene products as potential future contraceptive targets.