Function of MAP2D in Granulosa Cells

MAP2D 在颗粒细胞中的功能

• 批准号: 7386784
• 负责人: Mary E Hunzicker-Dunn
• 金额: $ 38.04万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-12-01 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7386784
• 关键词: A kinase anchoring protein; Abbreviations; Adaptor Signaling Protein; Affect; Affinity; Affinity Chromatography; Antibodies; Antisense Oligonucleotides; Appendix; Aromatase; Binding; Binding Proteins; Binding Sites; Biological Assay; CREB1 gene; Cell Proliferation; Cell surface; Cells; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cytochrome P450; Differentiation and Growth; Female; Fertility; Follicle Stimulating Hormone; G-Protein-Coupled Receptors; Genes; Growth; Growth Factor; Hand; Histone H3; Immunoprecipitation; In Vitro; Knockout Mice; Locales; Luteal Cells; Luteinization; Luteinizing Hormone; MEKKs; MEKs; Mass Spectrum Analysis; Measures; Mediating; Microtubule-Associated Proteins; Microtubules; Monomeric GTP-Binding Proteins; Oocytes; Ovarian; Ovarian Follicle; Ovulation; Pathway interactions; Peptides; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphorylation Site; Phosphotransferases; Pituitary Hormones; Procedures; Process; Proliferating; Protein Kinase A Inhibitor; Protein Tyrosine Phosphatase; Proteomics; Recombinant Inhibin A; Recombinants; Regulation; Research Personnel; Side; Signal Transduction; Signal Transduction Pathway; Signaling Protein; Site; Specificity; Staging; Surface; Testing; Thinking; Translating; Yeasts; granulosa cell; human INHA protein; human MAP3K1 protein; in vivo; inhibin; oocyte maturation; programs; protein expression; receptor; reproductive axis; response; scaffold; steroid hormone; uterine receptivity; yeast two hybrid system

DESCRIPTION (provided by applicant): Ovarian follicular maturation is a dynamic process that requires exquisite regulation. Follicles are restrained at the preantral (PA) stage until they are stimulated by follicle stimulating hormone (FSH). FSH signals by activating its receptor on the surface of granulosa cells via cAMP and its target protein kinase A (PKA). In response to FSH, granulosa cells proliferate and differentiate to a mature phenotype competent to respond to the preovulatory (PO) surge of luteinizing hormone (LH). LH then acts on these PO cells to activate PKA to promote terminal differentiation of granulosa into luteal cells, cessation of cell proliferation, and ovulation. While both FSH and LH signal to activate PKA, the pathways by which PKA directs responses are both similar and different in granulosa cells of PA versus PO follicles. For example, both FSH and LH promote histone H3 phosphorylation by a pathway that is blocked by the selective PKA inhibitor peptide myristoylated (Myr-) PKI. However, while both FSH and LH promote activation of ERK in a PKA-dependent manner (blocked by Myr- PKI), FSH does not promote activation of the upstream ERK kinase MEK in PA cells while LH promotes activation of MEK in PO cells. Additionally, there are numerous genes whose regulation by FSH in PA versus LH in PO cells is opposite, such as inhibin-alpha, aromatase, and LHR. The specificity of PKA action is thought to occur by the targeting of PKA and its substrates to specific cellular locales by virtue of its binding to A kinase anchoring proteins (AKAPs). We have shown that granulosa cells of PO follicles are distinguished from those of PA follicles by the presence of an 80 kDa AKAP that we recently identified as microtubule associated protein (MAP) 2D. We hypothesize that MAP2D functions as a signaling scaffold in PO granulosa cells to direct LH signals from PKA to a distinct set of substrates. Aim 1 will test the hypothesis that MAP2D functions as a scaffold to coordinate signal transduction pathways in PO granulosa cells; Aim 2 will test the hypothesis that MAP2D is obligatory for LH to signal to select pathways hi PO granulosa cells; Aim 3 will test the hypothesis that MAP2D is not associated microtubules and that the phosphorylation of MAP2D at specific sites regulates its association with PKA; and Aim 4 will test, the hypothesis that MAP2D is an Rl-preferential AKAP. Understanding how follicular maturation is regulated can translate into safer and more effective treatments for fertility.

描述（由申请人提供）：卵巢卵泡成熟是一个动态过程，需要精致的调节。卵泡在肌酸（PA）阶段受到约束，直到被卵泡刺激激素（FSH）刺激。 FSH信号通过CAMP及其靶蛋白激酶A（PKA）激活其受体在颗粒细胞表面上。为了响应FSH，颗粒细胞增殖并与成熟的表型相分化，能够响应黄体激素（LH）的排卵前（PO）激增。然后，LH作用于这些PO细胞以激活PKA，以促进颗粒末端分化为黄体细胞，停止细胞增殖和排卵。虽然FSH和LH信号都激活PKA，但PKA指导反应的途径在PA与PO卵泡的颗粒细胞中既相似又不同。例如，FSH和LH均通过通过选择性PKA抑制剂肽肉豆蔻酰化（MYR-）PKI阻断的途径促进组蛋白H3磷酸化。但是，尽管FSH和LH都以PKA依赖性方式促进ERK激活（由Myr-PKI阻止），但FSH并未促进PA细胞中上游ERK激酶MEK的激活，而LH促进PO细胞中MEK的激活。另外，有许多基因在PO细胞中对PA与LH的FSH调节相反，例如抑制蛋白-Alpha，芳香酶和LHR。 PKA作用的特异性被认为是通过PKA及其底物靶向特定细胞层的靶向，它通过其与激酶锚定蛋白（AKAP）的结合来实现。我们已经表明，Po卵泡的颗粒细胞通过存在80 kDa AKAP与Pa卵泡区分开，我们最近将其确定为微管相关蛋白（MAP）2D。我们假设MAP2D充当PO颗粒细胞中的信号支架，可将LH信号从PKA引导到一组不同的底物。 AIM 1将检验以下假设：MAP2D充当PO颗粒细胞中信号转导途径的支架。 AIM 2将检验以下假设：MAP2D必须强制LH发出信号以选择途径hi po granulosa细胞。 AIM 3将检验以下假设：MAP2D与微管无关，并且在特定位置的MAP2D磷酸化调节其与PKA的关联。 AIM 4将测试MAP2D是RL推荐AKAP的假设。了解如何调节卵泡成熟可以转化为更安全，更有效的生育能力。