Role of LH-induced cell migration and cofilin dephosphorylation in ovulation

LH 诱导的细胞迁移和丝动蛋白去磷酸化在排卵中的作用

基本信息

批准号：
10386571
负责人：
Corie Marie Owen
金额：
$ 4.04万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10386571
关键词：
Actins Antibodies Apical Basal lamina Biological Assay Blood Vessels Cell Count Cell Shape Cells Clinical Treatment Complex Disease Dominant-Negative Mutation Event Female infertility Fertility Gene Expression Glutamates Hemagglutinin Hour Image Immune Knowledge LH Receptors Label Lead Luteal Cells Luteinizing Hormone Mediator of activation protein Meiosis Methods Movement Mus Oocytes Ovary Ovulation Peptide Hydrolases Physiologic pulse Post-Translational Protein Processing Pregnancy Protein Dephosphorylation Proteins Receptor Signaling Regulation Research Proposals Role Rupture Serine Signal Transduction Site Surface Testing Time Tissues Woman actin depolymerizing proteins base cell motility cell type cofilin corpus luteum directional cell female fertility granulosa cell hormonal signals improved in vivo insight migration novel prevent receptor receptor expression response theca cell two photon microscopy

Actins Antibodies Apical Basal lamina Biological Assay Blood Vessels Cell Count Cell Shape Cells Clinical Treatment Complex Disease Dominant-Negative Mutation Event Female infertility Fertility Gene Expression Glutamates Hemagglutinin Hour Image Immune Knowledge LH Receptors Label Lead Luteal Cells Luteinizing Hormone Mediator of activation protein Meiosis Methods Movement Mus Oocytes Ovary Ovulation Peptide Hydrolases Physiologic pulse Post-Translational Protein Processing Pregnancy Protein Dephosphorylation Proteins Receptor Signaling Regulation Research Proposals Role Rupture Serine Signal Transduction Site Surface Testing Time Tissues Woman actin depolymerizing proteins base cell motility cell type cofilin corpus luteum directional cell female fertility granulosa cell hormonal signals improved in vivo insight migration novel prevent receptor receptor expression response theca cell two photon microscopy

展开

项目摘要

Project Abstract: In the ovary, luteinizing hormone (LH) signals through the luteinizing hormone receptor (LHR) to initiate the transition from preovulatory follicle to corpus luteum. Essential to this transition are the movement of the oocyte to the site of ovulation, basal lamina breakdown, and reorganization of the granulosa cells to form the corpus luteum. LHR-expressing cells migrate inwards in response to LH and at later time points after LH stimulation, the basal lamina is pulled inwards in regions of high HA-LHR expression. These results have led to the hypothesis that LH-induced granulosa cell migration aids in regulating the transition from preovulatory follicle to corpus luteum. Previous studies in isolated granulosa cells have indicated that LH signaling disrupts actin to allow cytoskeletal rearrangement and induces granulosa cell motility. One potential mediator of these actions is cofilin, an actin-depolymerizing protein. Cofilin activity is required for directional cell motility in multiple cell types, and expression of a dominant-negative form of the protein in granulosa cell inhibits LH-induced actin reorganization. However, the role of cofilin dephosphorylation in vivo has yet to be investigated. This project investigates the hypothesis that LH induces cell migration through dephosphorylation of cofilin, and that the migration pulls the basal lamina inward to aid in rupture during ovulation and reorganizes the cells to begin corpus luteum formation. In aim 1, live-tissue confocal and two-photon microscopy will be used to analyze granulosa cell migration in intact follicles and determine how granulosa cell migration contributes to basal lamina invaginations and ovulation. In aim two, a novel mouse line that expresses a dominant-negative form of cofilin will be used to explore the hypothesis that cofilin dephosphorylation regulates granulosa cell migration. These studies will provide novel information about the regulation of ovulation and could lead to clinical treatments for anovulatory diseases.

项目摘要：在卵巢中，通过黄体生成激素受体（LHR）启动黄体生成激素（LH）信号从卵泡前卵泡过渡到叶黄素。这种过渡至关重要的是卵母细胞的运动到排卵的部位，基底层分解和颗粒细胞的重组以形成语料库黄体。表达LHR的细胞会响应LH迁移，并在LH刺激后的以后时间迁移，在高HA-LHR表达的区域将基底层向内拉。这些结果导致了假设LH诱导的颗粒细胞迁移有助于调节从排卵前卵泡到 Luteum。先前在孤立的颗粒细胞中的研究表明，LH信号传导将肌动蛋白破坏为允许细胞骨架重排并诱导颗粒细胞运动。这些行动的一个潜在调解人是 cofilin，一种肌动蛋白 - 脱聚合蛋白。在多种细胞类型中定向细胞运动需要Cofilin活性，和颗粒细胞中蛋白质的显性阴性形式的表达抑制LH诱导的肌动蛋白重组。然而，cofilin deHosphoration在体内的作用尚未研究。这个项目研究了LH通过cofilin的去磷酸化诱导细胞迁移的假设，并且是迁移将基底层拉向内，以帮助排卵期间破裂，并重新组织细胞以开始叶黄素形成。在AIM 1中，将使用实时组织共聚焦和两光子显微镜进行分析完整卵泡中的颗粒细胞迁移，并确定颗粒细胞迁移如何有助于基底层令人讨厌和排卵。在AIM二，一种新型的小鼠线，表达了cofilin的主要阴性形式将用于探索Cofilin去磷酸化调节颗粒细胞迁移的假设。这些研究将提供有关排卵调节的新信息，并可能导致临床治疗发电性疾病。