Fertility and TAF4b: Transcriptional Regulation of Murine Ovarian Reserve Establishment

在美国，女性不孕不育是一个常见的健康问题。原发性卵巢功能不全(POI)以绝经提前为特征，导致1%的女性不孕，但在大多数情况下，不孕的原因无法确定。虽然POI通常在成年后被诊断，但这种情况很可能源于胚胎发育期间卵巢储备的建立不佳。卵巢储备由原始卵泡组成，每个原始卵泡包含一个被一层颗粒前细胞包围的单个卵母细胞。减数分裂和早期卵子发生是原始卵泡发育成败的两个关键因素，但对雌性生殖系中这些早期过程的转录调控知之甚少。我的研究重点是TBP相关因子4b(TAF4b)，它是一般转录机制中性腺富集亚单位，对女性生育至关重要。缺乏TAF4b的雌性小鼠表现出多方面的POI，并表现出减数分裂和卵子发生的严重缺陷。TAF4b缺乏对小鼠的影响可以追溯到减数分裂的启动，但TAF4b在这一过程中的功能仍然不清楚。这项建议的目的是全面了解TAF4b如何对驱动减数分裂基因表达和成功建立卵巢储备的基因调控网络做出贡献。目的1将使用一个强大的早期卵母细胞体外发育系统来阐明TAF4b在减数分裂启动和采用卵子起源命运中的确切作用。目标2将使用体内方法来评估TAF4b如何以及何时与其他调节减数分裂和卵子发生的因素相互作用。最终，这项建议将阐明一般转录机制的组成部分如何调节对减数分裂和卵子发生的正常进行至关重要的基因表达，从而将显著有助于该领域对女性生殖细胞发育的理解。在完成这些目标的同时，我将发展一套在生殖生物学研究领域取得成功所需的技能和关键基础知识。通过参加伍兹霍尔的生殖前沿课程，在国内和国际会议上发表演讲，以及我的赞助商的深思熟虑的指导，我的培训经验将得到丰富。此外，这项提案将在布朗大学杰出的支持性分子生物学、细胞生物学和生物化学研究生项目中完成。这一奖学金的完成将推动发育生物学领域的发展，并为我实现领导一个专注于生殖生物学的独立学术研究实验室的职业目标提供所需的特殊准备。

Female infertility is a common health concern in the United States. Primary ovarian insufficiency (POI) is characterized by premature menopause and it leads to infertility in 1% of the female population, but the cause of infertility cannot be determined in most cases. Although POI is often diagnosed in adult life, this condition likely stems from poor establishment of the ovarian reserve during embryonic development. The ovarian reserve is composed of primordial follicles, each of which contain a single oocyte surrounded by a layer of pre-granulosa cells. Meiosis and early oogenesis are two key contributors to the success or failure of primordial follicle development, but the transcriptional controls regulating these early processes in the female germline are poorly understood. The focus of my research is TBP-Associated factor 4b (TAF4b), which is a gonadally-enriched subunit of the general transcription machinery and it is essential for female fertility. Female mice lacking TAF4b display multiple aspects of POI and exhibit critical defects in meiosis and oogenesis. The effects of Taf4b- deficiency in the mouse can be traced back to meiotic initiation, but the function of TAF4b during this process has remained elusive. The goal of this proposal is to develop a comprehensive understanding of how TAF4b contributes to the gene regulatory network that drives expression of meiotic genes and successful establishment of the ovarian reserve. Aim 1 will use a powerful in vitro system of early oocyte development to elucidate the precise role of TAF4b in meiotic initiation and adoption of the oogenic fate. Aim 2 will use an in vivo approach to evaluate how and when TAF4b interacts with other factors that regulate meiosis and oogenesis. Ultimately, this proposal will clarify how components of the general transcription machinery regulate gene expression that is essential for proper progression of meiosis and oogenesis, and thus it will significantly contribute to the field’s understanding of female germ cell development. While completing these aims, I will develop a repertoire of skills and the key foundational knowledge required for a successful career in reproductive biology research. My training experience will be enriched by attending the Frontiers in Reproduction course in Woods Hole, presenting at national and international conferences, and thoughtful mentoring by my sponsor. Moreover, completion of this proposal will take place in the outstandingly supportive Molecular Biology, Cell Biology, and Biochemistry Graduate Program at Brown University. Completion of this fellowship will move the developmental biology field forward and provide me with the exceptional preparation required for the achievement of my career goal to lead an independent academic research laboratory focused on reproductive biology.