Oocyte stage-specific role of ERβ in primordial follicle activation

ERβ 在原始卵泡激活中的卵母细胞阶段特异性作用

• 批准号: 10493320
• 负责人: Mohammad A.K. Rumi
• 金额: $ 19.38万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10493320
• 关键词: Assisted Reproductive Technology; Bioinformatics; Cells; ChIP-seq; Clinical Trials; DNA Binding Domain; Development; Ensure; Estrogen Receptor beta; Estrogen Receptors; Female; Gatekeeping; Genes; Genetic Transcription; Knock-out; Knockout Mice; Life; Ligands; Longevity; Mediating; Modeling; Molecular; Molecular Target; Mus; Neonatal; Nuclear Receptors; Oocytes; Oogonia; Ovarian; Ovarian Follicle; Ovary; Phenotype; Play; Primordial Follicle; Process; Rattus; Regulation; Research; Role; Signal Transduction; Signaling Molecule; Source; Testing; Therapeutic Trials; Transcriptional Regulation; base; conditional knockout; fetal; granulosa cell; improved; innovation; insight; mouse model; mutant; neonatal period; novel; novel strategies; ovarian dysfunction; ovarian reserve; premature; prenatal; preservation; prevent; reproductive; reproductive longevity; single-cell RNA sequencing; transcriptome

SUMMARY In mammalian ovaries, a fixed number of primordial follicles that are formed during the fetal or early neonatal period serve as the source of mature oocytes for the entire reproductive lifespan. Uncontrolled or excessive activation of primordial follicles leads to the depletion of the ovarian reserve, resulting in ovulatory dysfunction. We demonstrated that estrogen receptor β (ERβ) plays a gatekeeping role during primordial follicle activation (PFA). In both the rat and the mouse, the absence of ERβ results in increased numbers of primordial follicles being activated and developing into primary follicles. In a conditional knockout mouse that lacks ERβ in oocytes starting from the primordial follicle stage, we detected excessive activation of primordial follicles similar to that of Erβnull mutants. Based on this novel observation, we hypothesize that ERβ regulation of PFA is mediated by its transcriptional function in the oocytes of primordial or primary follicles. However, it remains unclear if the regulatory mechanism of ERβ is restricted to the primordial follicles or if it can function in activated primary follicles. Also, our observations cannot exclude a potential role for ERβ in follicular granulosa cells. In this study, we propose to define oocyte stage-specific roles of ERβ in the regulation of PFA. Using follicular stage-specific conditional ERβ knockout mice, we will: 1) determine the role of ERβ in controlling PFA in both primordial and activated oocytes, 2) assess the ovarian phenotypes following oocyte stage-specific ERβ depletion, 3) perform single cell transcriptome analyses to identify the ERβ-regulated oocyte genes involved in PFA, and 4) identify any granulosa-specific genes that are differentially impacted due to oocyte-specific depletion of ERβ. ERβ is a ligand activated nuclear receptor that has well-characterized selective ligands approved for use in clinical trials. Although it is the predominant estrogen receptor in the ovary, ERβ has never been targeted to treat ovarian dysfunction. The results of the proposed research will provide insight into how the molecular targeting of ERβ might enhance our efforts to extend female reproductive longevity and improve existing assisted reproductive technologies.

总结 在哺乳动物卵巢中，在胎儿期或新生儿早期形成的固定数量的原始卵泡 作为整个生殖寿命的成熟卵母细胞的来源。不受控制或过度激活 原始卵泡导致卵巢储备耗竭，导致排卵功能障碍。我们 证明雌激素受体β（ERβ）在原始卵泡激活（PFA）过程中发挥守门作用。 在大鼠和小鼠中，ERβ的缺乏导致原始卵泡数量的增加， 被激活并发育成初级卵泡。在卵母细胞中缺乏ERβ的条件性敲除小鼠中， 从原始卵泡阶段开始，我们检测到与Erβnull相似的原始卵泡过度激活 变种人基于这一新的观察结果，我们假设PFA的ERβ调节是由其 在原始或初级卵泡的卵母细胞中的转录功能。然而，目前尚不清楚， ERβ的调节机制仅限于原始卵泡或是否能在激活的初级卵泡中发挥作用。 此外，我们的观察不能排除ERβ在卵泡颗粒细胞中的潜在作用。在这项研究中，我们建议 明确ERβ在卵母细胞PFA调控中的阶段特异性作用。使用卵泡阶段特异性条件ERβ 敲除小鼠，我们将：1）确定ERβ在控制原始卵母细胞和激活卵母细胞中PFA的作用，2） 评估卵母细胞阶段特异性ERβ耗竭后的卵巢表型，3）进行单细胞转录组 分析以鉴定参与PFA的ERβ调节的卵母细胞基因，和4）鉴定任何颗粒体特异性基因 由于ERβ的卵母细胞特异性消耗而受到不同影响。ERβ是一种配体激活的核受体 其具有被批准用于临床试验的充分表征的选择性配体。虽然它是主要的 ER β是卵巢中的雌激素受体，ERβ从未被靶向治疗卵巢功能障碍。的结果 拟议的研究将提供深入了解ERβ的分子靶向如何增强我们的努力， 女性生殖寿命和改善现有的辅助生殖技术。