BFGF AND CELL CONTACT REGULATE GRANULOSA CELL APOPTOSIS

描述(改编自《调查员摘要》)：该提案是 分为7个具体目标。该提案的总体目标是 确定控制颗粒细胞活性的细胞机制。这个 研究人员产生的初步数据表明，碱性成纤维细胞 生长因子，通过旁分泌或自分泌机制和细胞 接触相互作用，调节颗粒细胞的活性。蜂窝触点 这一机制的组成部分似乎是通过N-钙粘附素而不是通过 缝隙连接蛋白。有趣的是，调查人员产生了 初步数据表明N-钙粘附素与受体相互作用 碱性成纤维细胞生长因子这一观察结果表明，黏附分子可能调节细胞 通过生长因子受体改变信号转导的生存能力。 这是生物学中一个有趣的普遍原则，值得更多的关注。 详细调查。 如上所述，该提案分为7个具体目标。具体而言 目的1，研究人员将确定一种合成肽(LRAHAVDVNG)， 来源于在胞外区发现的氨基酸序列 成纤维细胞生长因子受体和N-钙粘附素的表达减弱碱性成纤维细胞生长因子诱导 颗粒细胞聚集，维持颗粒细胞活力。在……里面 具体目标2，研究人员计划确定N-钙粘附素 抗体阻止碱性成纤维细胞生长因子与其受体结合。在具体目标3中， 研究人员将确定碱性成纤维细胞生长因子是否与其受体结合并导致 形成N-钙粘蛋白/成纤维细胞生长因子受体复合体。在具体目标4中， 研究人员将确定N-钙粘附素抗体是否会削弱bFGF的能力 刺激其受体的酪氨酸磷酸化以及其他 蛋白质。在特定的目标5中，研究人员将确定N-钙粘附素 抗体破坏颗粒细胞的肌动蛋白细胞骨架，从而 在不改变蛋白酪氨酸或在改变蛋白酪氨酸之前诱导细胞凋亡 磷酸化。在具体目标6中，调查者将确定碱性成纤维细胞生长因子 减少颗粒细胞凋亡并诱导颗粒细胞聚集 调节酪氨酸磷酸化状态和/或细胞的表达 N-钙粘附素及其相关连环蛋白的定位。在具体目标7中， 研究人员将测定碱性成纤维细胞生长因子及其受体的蛋白水平， 闭锁卵泡和非闭锁卵泡中N-钙粘附素和连环素的表达。这个 研究人员希望，这些研究将提供对 碱性成纤维细胞生长因子与颗粒细胞相互作用的细胞机制 调节颗粒细胞活性。

DESCRIPTION (Adapted from the Investigator's Abstract): The proposal is divided into 7 specific aims. The overall goal of the proposal is to identify the cellular mechanisms that control granulosa cell viability. The investigator has generated preliminary data that both basic fibroblast growth factor, acting through a paracrine or autocrine mechanism and cell contact interact to regulate granulosa cell viability. The cell contact component of this mechanism appears to be mediated via N--cadherin, not by gap junction proteins. Interestingly, the investigators have generated preliminary data which suggests N-cadherin interacts with the receptor for bFGF. This observation suggests that adhesion molecules may modulate cell viability by altering signal transduction through growth factor receptors. This is an interesting general principle in biology which deserves more detailed investigation. As noted above, the proposal is divided into 7 specific aims. In Specific Aim 1, the investigator will determine if a synthetic peptide (LRAHAVDVNG), derived from an amino acid sequence found within the extracellular domains of both the FGF receptor and N-cadherin attenuates bFGF's ability to induce granulosa cell aggregation and maintain granulosa cell viability. In Specific Aim 2, the investigator plans to determine if the N-cadherin antibody prevents bFGF from binding to its receptor. In Specific Aim 3, the investigator will determine if bFGF binds to its receptor and causes the formation of an N-cadherin/FGF receptor complex. In Specific Aim 4, the investigator will determine if N-cadherin antibody attenuates bFGF's ability to stimulate tyrosine phosphorylation of its receptor as well as other proteins. In Specific Aim 5, the investigator will determine if N-cadherin antibody disrupts the actin cytoskeleton of the granulosa cell and thereby induces apoptosis without or prior to altering protein tyrosine phosphorylation. In Specific Aim 6, the investigator will determine if bFGF reduces granulosa cell apoptosis and induces granulosa cell aggregation by regulating the expression of tyrosine phosphorylation status and/or cellular localization of N-cadherin and the associated catenins. In Specific Aim 7, the investigator will determine the protein levels of bFGF, FGF receptor, N-cadherin, and catenins in atretic and non-atretic follicles. The investigator hopes that these studies will provide new insights into the cellular mechanism through which bFGF and granulosa contact interact to regulate granulosa cell viability.