ACTIONS OF BONE MORPHOGENETIC PROTEINS IN THE PRIMATE OVARY

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。本研究的长期目标是了解骨形态发生蛋白（BMPs）在控制卵巢功能中的作用，特别是在灵长类动物中卵泡发育和卵母细胞成熟。来自啮齿类动物的证据表明，bmp，特别是GDF-9和BMP-15，具有重要的局部作用，控制卵泡/颗粒细胞分化和卵母细胞发育，包括排卵前卵丘-卵母细胞复合物的扩张。由贝勒大学（M. Matzuk）和俄勒冈健康与科学大学（R. Stouffer） NICHD生殖研究专业合作中心项目产生的初步数据表明，灵长类动物卵巢中存在并活跃着GDF-9/ bmp -15受体系统。因此，翻译研究计划如下：(1)生产大量纯化的重组人GDF-9和BMP-15，以及针对这些蛋白质的中和抗体，用于灵长类动物的体外和体内研究；(2)研究外源性和内源性GDF-9和BMP-15对非人灵长类动物和女性壁粒细胞和卵母细胞复合物（COCs）的影响。Matzuk博士的团队将在Aim 1进行实验，并在Aim 2进行人体细胞实验。Stouffer博士的团队将在Aim No. 2中使用恒河猴细胞进行实验：(a)将在存在和不存在bmp的情况下培养来自排卵前卵泡的猕猴颗粒细胞，并将RNA制剂运往贝勒大学，通过微阵列分析转录组；(b)猕猴积云卵母细胞复合物将在存在或不存在促性腺激素、BMP和/或BMP抗体的情况下孵育，并通过图像分析定量积云扩增。

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 long-range goal of this research is to understand the role of bone morphogenetic proteins (BMPs) in controlling ovarian function, particularly follicular development and oocyte maturation in primates. Evidence from rodents suggests that BMPs, notably GDF-9 and BMP-15, have important local actions controlling follicular/granulosa cell differentiation and oocyte development, including expansion of the cumulus-oocyte-complex just prior to ovulation. Preliminary data generated by the NICHD's Specialized Cooperative Centers Program in Reproduction Research at Baylor University (M. Matzuk) and Oregon Health & Science University (R. Stouffer) suggest that the GDF-9/BMP-15-receptor systems are present and active in the primate ovary. Therefore, translational studies are planned: (1) To produce large amounts of purified recombinant human GDF-9 and BMP-15, plus neutralizing antibodies to these proteins, for in vitro and in vivo studies in primates; and (2) To examine the effects of exogenous and endogenous GDF-9 and BMP-15 on mural granulosa cells and cumulus-oocyte-complexes (COCs) from nonhuman primates and women. Dr. Matzuk's group will perform experiments in Aim No. 1, as well as experiments with human cells in Aim No. 2. Dr. Stouffer's group will perform experiments using rhesus monkey cells in Aim No. 2: (a) macaque granulosa cells from preovulatory follicles will be cultured in the presence and absence of BMPs, and RNA preparations shipped to Baylor for analysis of the transcriptomes by microarrays; (b) macaque cumulus-oocyte-complexes will be incubated in the presence or absence of gonadotropin hormones, BMPs and/or BMP antibodies, and cumulus expansion quantitated by image analysis.