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。 这项补助金的目的是探索的假设，减数分裂激活和抑制基因的选择性表达存在于灵长类动物卵巢，并候选分子可以测试避孕潜力，使用一个既定的序列，在体外和体内实验最终在一个避孕试验中的猕猴繁殖组。 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;并研究所选候选物（例如，INSL 3-LGR 8途径、WEE 1B和卵母细胞特异性NLRP）的体外功能，（2）确定所选候选物是否可在恒河猴体内用作避孕剂，和（3）确定所选候选物（例如，INSL 3）可以在配对交配情况下在定期循环的恒河猴中作为避孕剂起作用。 实验设计将包括：基因产物表征（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）;在控制性卵巢刺激（COS）和自发月经周期期间向猴体内施用INSL 3或其拮抗剂，加上内分泌和毒性测量（目标2）;以及向繁殖群体中的可育雌性长期施用INSL 3或其抑制剂以评估避孕功效和长期毒性（目的3）。 这种方法有望为将来在人类中进行INSL 3试验提供基础，并确定其他卵母细胞特异性新基因产物作为未来潜在的避孕靶点。

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.