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: 10300681
• 负责人: Jennifer C McKey
• 金额: $ 10.34万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-09 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10300681
• 关键词: Ablation; Adipose tissue; Adult; Advisory Committees; Affect; Anatomy; Biological; Biological Process; Cells; Cues; Data; Development; Developmental Biology; Dextrans; Diet; Endocrine; Epithelial; 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; 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 aging; Ovary; Ovulation; Pattern; Physiological; Play; Primordial Follicle; Process; Regulation; Reproductive Biology; Research; Role; Running; Sheep; 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; 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。 女性生殖衰老的特征是由于 构成有限卵巢储备的卵巢卵泡数量和质量的顺序减少。这 rete ovarii（ro）是一种保守的上皮结构，分为3个区域，即沃里安（IOR），位于 在卵巢中，位于外周日组织中的外洛瓦里亚人（EOR）和连接（Cr）， 链接了eor和ior。在解剖环境中，RO已被描绘了数十年，这是一种可能的 导致妇科疾病的贡献者，但其功能从未得到彻底研究。生殖 女性的寿命由三个主要因素确定：1）初始原始卵泡池的大小，2） 肥沃的窗口期间排卵率和闭锁速率，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。此应用的目的是 阐明RO对这些决定因素的贡献。允许可视化和 卵巢原位的功能研究，我开发了组织清除和3D成像方法 rete在卵巢细胞组装中的作用 髓质和皮质域。我发现RO在整个卵巢中仍然与卵巢密切相关 成年和初步的卵巢移植实验表明，宿主EOR/CR重新建立了A 与移植卵巢的IOR连接。我们的RO的转录组数据显示了分泌的丰富 与注入EOR的葡聚糖的发现相一致的代谢途径被转运到 子房。这些数据导致了一个中心假设，即RO在机构中起着关键作用 卵巢储备和对生理提示的卵巢卵泡生长的调节。 该假设将通过追求三个具体目标来检验：（1）（K99）表征了该假设的贡献。 卵巢开发和卵巢储备的建立； （2）（K99/r00）定义 EOR与IOR之间的功能差异，（3）（R00）研究的变化 RO响应生理提示。发现ovarii在建立中的作用 卵巢储备和在稳态或扰动条件下对成年卵泡生长的调节 将把RO作为新候选人寻找女性生育和生殖的决定因素 长寿。此外，我开发的方法将在以后对互动的调查中很有用 在卵巢和其他外部组织之间。追求这些目标需要在单细胞RNA中进行培训 测序，小鼠手术和病毒介导的方法来操纵基因表达。我有 组建了一个由Capel博士主持的咨询委员会，我们设计了一个培训计划，将为我提供 拥有经营富有成效的，独立的发展和生殖生物学实验室所需的技能。