Functions of the rete ovarii in ovary development, adult homeostasis, and female reproductive longevity

卵巢网在卵巢发育、成人体内平衡和女性生殖寿命中的功能

• 批准号: 10828038
• 负责人: Jennifer C McKey
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-15 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10828038
• 关键词: Ablation; Adipose tissue; Adult; Advisory Committees; Affect; Anatomy; Biological; Biological Process; Cells; Cues; Data; Development; Developmental Biology; Dextrans; Diet; Endocrine; Epithelium; Female; Female Genital Diseases; Fertile Period; Fertility; Fetal Development; Future; Gene Expression; Genes; Genetic; Goals; Growth; Homeostasis; Hormones; Image; Immune; In Situ; Investigation; Knowledge; Laboratories; Life; Ligation; Light; Link; Location; Longevity; Lymphatic; Mediating; Medulla of ovary; Meiosis; Metabolic; Metabolic Pathway; Metabolic stress; Metabolism; Methods; Microscopy; Modeling; Molecular; Molecular Profiling; Mus; Nature; Nerve; Neuropeptides; Oocytes; Operative Surgical Procedures; Ovarian; Ovarian Follicle; Ovarian Tissue; Ovarian aging; Ovary; Ovulation; Pattern; Physiological; Play; Primordial Follicle; Process; Productivity; Regulation; Reproductive Biology; Research; Role; Running; Sheep; Specific qualifier value; Stromal Cells; Structure; Testing; Three-Dimensional Image; Three-Dimensional Imaging; Tissues; Training; Viral; Virus; Visualization; body system; design; experimental study; female fertility; fetal; gene function; genetic manipulation; imaging modality; in vivo; intraovarian; mouse model; ovarian reserve; ovary transplantation; postnatal; reproductive; reproductive function; reproductive longevity; reproductive senescence; response; single-cell RNA sequencing; skills; tool; transcriptomics

Abstract There is an urgent need to understand the factors determining female reproductive longevity. Female reproductive aging is characterized by a significant decline in reproductive function due to the sequential decrease in the number and quality of ovarian follicles that constitute the finite ovarian reserve. The rete ovarii (RO) is a conserved epithelial structure divided into 3 regions, the intraovarian rete (IOR), located within the ovary, the extraovarian rete (EOR), located in the periovarian tissue, and the connecting rete (CR), which links the EOR and IOR. The RO has been depicted for decades in anatomical context, and is a possible contributor to gynecological disease, yet its function has never been thoroughly investigated. The reproductive longevity of females is determined by three main factors: 1) the size of the initial primordial follicle pool, 2) the rate of ovulation and atresia during the fertile window, and 3) perturbations, such as changes in metabolic status or hormone levels, which affect ovarian function and follicle growth. The objective of this application is to elucidate the contribution of the RO to each of these determining factors. To allow visualization and functional investigation of the ovary in situ, I developed tissue clearing and 3D imaging methods that suggest a role for the rete in the assembly of ovarian cells into ovigorous cords and subsequent specification of medullary and cortical domains. I found that the RO remains closely associated with the ovary throughout adulthood, and preliminary ovary transplantation experiments suggest that the host EOR/CR reestablishes a connection to the IOR of the grafted ovary. Our transcriptomic data of the RO shows enrichment of secretory and metabolic pathways consistent with the finding that Dextran injected into the EOR is transported to the ovary. These data have led to the central hypothesis that the RO plays critical roles in the establishment of the ovarian reserve and in the regulation of ovarian follicle growth in response to physiological cues. This hypothesis will be tested by pursuing three specific aims: (1) (K99) Characterize the contribution of the RO to the development of the ovary and the establishment of the ovarian reserve; (2) (K99/R00) Define the functional differences between EOR and IOR in adult homeostasis and (3) (R00) Investigate the changes of the RO in response to physiological cues. Discovering a role for the rete ovarii in the establishment of the ovarian reserve and in the regulation of adult follicle growth under homeostatic or perturbed conditions will situate the RO as a new candidate in the search for determinants of female fertility and reproductive longevity. In addition, the approaches I develop will be useful in future investigations of interactions between the ovary and other extrinsic tissues. Pursuit of these aims requires training in single-cell RNA sequencing, mouse surgery and virally-mediated approaches for manipulating gene expression. I have assembled an advisory committee chaired by Dr. Capel, and we designed a training plan that will provide me with the skills required to run a productive, independent developmental and reproductive biology laboratory.

抽象的 迫切需要了解决定女性生殖寿命的因素。 女性生殖衰老的特点是生殖功能显着下降。 构成有限卵巢储备的卵巢卵泡的数量和质量连续下降。这 卵巢网 (RO) 是一种保守的上皮结构，分为 3 个区域，即卵巢内网 (IOR)，位于 在卵巢内，位于卵巢周围组织的卵巢外网（EOR）和连接网（CR）， 它链接 EOR 和 IOR。 RO 几十年来一直在解剖学背景下被描述，并且是一个可能的 是导致妇科疾病的一个因素，但其功能尚未得到彻底研究。生殖的 女性的寿命由三个主要因素决定：1）初始原始卵泡池的大小，2） 受孕窗口期间的排卵率和闭锁率，以及 3) 扰动，例如代谢变化 状态或激素水平，影响卵巢功能和卵泡生长。该应用程序的目标是 阐明 RO 对每个决定因素的贡献。为了允许可视化和 为了对卵巢进行原位功能研究，我开发了组织透明化和 3D 成像方法，表明 rete 在卵巢细胞组装成产卵索和随后的规格中的作用 髓质和皮质域。我发现 RO 始终与卵巢密切相关 成年期和初步卵巢移植实验表明宿主 EOR/CR 重新建立了 与移植卵巢 IOR 的连接。我们的 RO 转录组数据显示分泌丰富 和代谢途径与注入 EOR 的葡聚糖被转运到 子房。这些数据得出了一个中心假设：RO 在企业建立过程中发挥着关键作用。 卵巢储备功能以及根据生理信号调节卵泡生长。 该假设将通过追求三个具体目标来检验：(1) (K99) 描述 RO对卵巢的发育和卵巢储备功能的建立； (2) (K99/R00) 定义 EOR 和 IOR 在成人稳态中的功能差异以及 (3) (R00) 研究 RO 对生理信号做出反应。发现卵巢网在建立 卵巢储备功能以及在稳态或扰动条件下调节成年卵泡的生长 将把 RO 作为寻找女性生育力和生殖决定因素的新候选人 长寿。此外，我开发的方法将有助于未来的相互作用研究 卵巢和其他外在组织之间。追求这些目标需要单细胞 RNA 培训 测序、小鼠手术和病毒介导的基因表达方法。我有 组建了一个由卡佩尔博士担任主席的咨询委员会，我们设计了一个培训计划，该计划将为我提供 具备运行高效、独立的发育和生殖生物学实验室所需的技能。