Cyclic AMP Signaling in Granulosa Cells

颗粒细胞中的环 AMP 信号转导

基本信息

批准号：
8485626
负责人：
Anthony J Zeleznik
金额：
$ 25.78万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-15 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8485626
关键词：
A kinase anchoring protein AKAP13 gene Accounting Address Adenoviruses Affect Aromatase Biosensor Cell Differentiation process Cell Extracts Cell Nucleus Complex Contraceptive Agents Coupling Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cytoplasm Development Disease Dose Estradiol Estrogens Fertility Agents Fluorescence Resonance Energy Transfer Follicle Stimulating Hormone Forskolin Gene Expression Gene Expression Profile Genes Goals Immunoblot Analysis LH Receptors Lentivirus Vector Luteinization Monitor Ovarian Ovarian Follicle Ovulation Pathway interactions Peptides Process Production Progesterone Regulation Reporter Reporting Research Role Signal Pathway Signal Transduction Study Section Syndrome System Testing Time Undifferentiated Viral Vector Western Blotting base folliculogenesis granulosa cell mRNA Expression novel ovarian failure public health relevance reproductive success response sensor transcription factor

项目摘要

DESCRIPTION (provided by applicant): Although progress has been made in understanding the role of follicle stimulating hormone (FSH) in governing the process of preovulatory folliculogenesis, a major question that remains unanswered is the elucidation of the intracellular signaling pathways utilized by FSH in the regulation of the complex pattern of gene expression that occurs during follicular development. Our preliminary results point to two loci through which FSH may optimally induce granulosa cell differentiation. First, activation of a pathway or pathways in addition to PKA synergizes with the PKA pathway to optimally induce the expression of a subset of genes including aromatase and the LH receptor, the two hallmark genes associated with granulosa cell differentiation. Second, compartmentalization of the FSH/cAMP signaling pathway may result in more efficient coupling of intracellular cAMP production with the induction of the estrogen and progesterone biosynthetic pathways. In this current proposal we present four Specific Aims to address these findings. Aim 1 will compare the signaling pathways activated by FSH and by PKA-CQR in undifferentiated granulosa cells. Aim 2 will determine which signaling pathways and/or transcription factors interact with PKA to optimally induce granulosa cell differentiation. Aim 3 will explore the possible intracellular compartmentalization of the cAMP/PKA signaling system in granulosa cells by disrupting PKA/AKAP interactions with HT31 peptides on granulosa cell differentiation and using novel FRET-based PKA sensors to directly evaluate compartmentalization of cAMP/PKA signaling in granulosa cells. Understanding the signaling pathways involved in granulosa cell differentiation may be instrumental for the development of novel contraceptives and "fertility drugs" as well as providing new information regarding signaling pathways that may be affected in anovulatory disorders such as ovarian failure and polycystic ovarian syndrome.

描述（由申请人提供）：尽管在了解促卵泡激素（FSH）在控制排卵前卵泡发生过程中的作用方面已经取得了进展，但仍未得到解答的一个主要问题是阐明 FSH 在排卵前卵泡发生过程中所利用的细胞内信号传导途径。卵泡发育过程中发生的复杂基因表达模式的调节。我们的初步结果指出 FSH 可通过两个位点最佳地诱导颗粒细胞分化。首先，除PKA之外的一个或多个途径的激活与PKA途径协同作用，以最佳地诱导包括芳香酶和LH受体在内的基因子集的表达，这两个基因与颗粒细胞分化相关。其次，FSH/cAMP 信号通路的区室化可能导致细胞内 cAMP 产生与雌激素和孕激素生物合成通路的诱导更有效地耦合。在当前的提案中，我们提出了四个具体目标来解决这些发现。目标 1 将比较未分化颗粒细胞中 FSH 和 PKA-CQR 激活的信号传导途径。目标 2 将确定哪些信号通路和/或转录因子与 PKA 相互作用以最佳地诱导颗粒细胞分化。目标 3 将通过破坏 PKA/AKAP 与 HT31 肽对颗粒细胞分化的相互作用，并使用新型基于 FRET 的 PKA 传感器直接评估颗粒细胞中 cAMP/PKA 信号传导的区室化，探索颗粒细胞中 cAMP/PKA 信号传导系统可能的细胞内区室化细胞。了解颗粒细胞分化所涉及的信号通路可能有助于开发新型避孕药和“生育药物”，并提供有关可能受到卵巢功能衰竭和多囊卵巢综合征等无排卵疾病影响的信号通路的新信息。