Anti-Mullerian Hormone Actions to Control Primate Folliculogenesis

项目总结 抗苗勒氏激素(AMH)在卵泡发育过程中起着关键作用。我们已经推进了 我们对AMH的动态表达、直接作用和生理作用的理解 自该项目最初获奖以来，恒河猴的卵泡发生。数据表明，AMH是由 通过发育卵泡异质性，并通过促进腔前表现出阶段依赖的调节效应 卵泡生长和防止有腔卵泡成熟。阶段特异性AMH调节促进卵泡生长 并在体外成熟以产生合格的卵母细胞。因此，进一步的研究是有必要澄清的。 AMH对卵泡发育特定阶段下游分子和细胞事件的调控 发育以及调节AMH作用以改善卵巢功能和女性的可能性 生育能力。我们的初步研究表明：(A)体内应用AMH可促进腔前卵泡生长， 导致卵巢中有腔卵泡数量的增加；(B)原始卵泡的激活和生长 腔前卵泡是通过冷冻保存后的卵巢组织培养获得的；以及(C)腔前卵泡 维持在生理缺氧的状态。AMH有可能通过以下方式调节细胞周期进程 限制缺氧诱导因子的作用，并通过诱导肌酸来控制细胞能量的产生 低氧条件下的生物合成和代谢。因此，将进行研究，以检验阶段- AMH作用的特定调节促进了体内和体外的卵泡发育，进而改善了卵泡的发育 生育结果。目的1将评估AMH预充对促性腺激素卵巢反应的影响 体外受精(体外受精)中的刺激。AMH将在晚期猕猴体内给药 育龄或对控制性卵巢刺激(COS)反应差。然后COS协议将被 进行体外受精效果评估。目的2将检测AMH对体外成熟的影响。 来自冷冻猕猴和人类卵巢组织的卵泡。玻璃化加温的卵巢皮质将被 培养诱导原始卵泡激活和生长至腔前阶段。然后腔前卵泡将被 分离培养，用AMH调节卵母细胞成熟。目标3将确定分子 AMH在卵泡发育过程中调节细胞增殖和能量产生的机制。 猕猴腔前卵泡将在5%O2中添加AMH或击倒培养。因素 将分析调节细胞周期进程和肌酸生物合成/代谢途径的组成部分。 拟议的研究将为AMH的阶段依赖性作用机制提供重要的见解。 在调节卵泡发育和卵巢功能方面，使用非人类灵长类动物模型是一种极好的 了解女性生殖过程的替代者。这些发现将使开发协议成为可能 用于优化试管受精过程，改进女性生育能力保存的选择，并定义AMH介导的 卵泡发生过程中细胞周期和能量代谢的调节。

PROJECT SUMMARY Anti-Müllerian hormone (AMH) plays a key role during ovarian follicular development. We have advanced our understanding of the dynamic expression, direct action, and physiological role of AMH during folliculogenesis in rhesus macaques since the initial award of the project. Data suggest that AMH is produced heterogeneously by developing follicles and exhibits stage-dependent regulatory effects by promoting preantral follicle growth and preventing antral follicle maturation. Stage-specific AMH modulation improves follicle growth and maturation in vitro to generate competent oocytes. Therefore, further research is warranted to elucidate the downstream molecular and cellular events regulated by AMH at the specific stages of follicular development, as well as the potential for modulation of AMH actions to improve ovarian function and female fertility. Our preliminary studies show that (a) AMH administration in vivo promotes preantral follicle growth, resulting in the increased number of antral follicles in the ovary; (b) primordial follicle activation and growth to the preantral stage is achieved by ovarian tissue culture after cryopreservation; and (c) preantral follicles are maintained in a state of physiological hypoxia. AMH has the potential to regulate cell cycle progression by limiting actions of hypoxia-inducible factor, and to control cellular energy production by inducing creatine biosynthesis and metabolism under hypoxia. Thus, studies will be performed to test the hypothesis that stage- specific modulation of AMH actions facilitates follicular development in vivo and in vitro, which in turn improves fertility outcomes. Aim 1 will evaluate the impact of AMH priming on ovarian response to gonadotropin stimulation during IVF (in vitro fertilization). AMH will be administered in vivo in macaques at advanced reproductive age or with poor response to controlled ovarian stimulation (COS). The COS protocol will then be performed to evaluate IVF efficacy. Aim 2 will examine the effect of AMH modulation on in vitro maturation of follicles from cryopreserved macaque and human ovarian tissue. Vitrified-warmed ovarian cortex will be cultured to induce primordial follicle activation and growth to the preantral stage. Preantral follicles will then be isolated and cultured with AMH modulation for oocyte maturation. Aim 3 will determine the molecular mechanisms whereby AMH regulates cell proliferation and energy production during follicular development. Macaque preantral follicles will be cultured at 5% O2 with AMH supplementation or knockdown. Factors regulating cell cycle progression and creatine biosynthesis/metabolic pathway components will be analyzed. The proposed studies will provide important insight into the mechanism of stage-dependent AMH actions in regulating follicular development and ovarian function using a nonhuman primate model that is an excellent surrogate for understanding reproductive processes in women. The findings will allow for developing protocols for optimizing IVF process, advancing options for female fertility preservation, and defining AMH-mediated regulation of cell cycle and energy metabolism during folliculogenesis.