A PR-Induced Autocrine/Paracrine Pathway in Female Brain

PR 诱导女性大脑中的自分泌/旁分泌途径

• 批准号: 6467321
• 负责人: Ede M Apostolakis
• 金额: $ 23.7万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-12 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6467321
• 关键词: adenylate cyclase; autocrine; binding proteins; biological signal transduction; cyclic AMP; estrogen receptors; estrogens; female; gene expression; hormone regulation /control mechanism; hypothalamus; immunocytochemistry; in situ hybridization; laboratory mouse; laboratory rat; microarray technology; mitogen activated protein kinase; neurons; neuropeptide receptor; neuropeptides; paracrine; peptide hormone biosynthesis; polymerase chain reaction; progesterone; progesterone receptors; receptor expression; receptor sensitivity

Ovarian steroid hormones, estrogen (E) and progesterone (P), exert profound influence on development, reproduction, and aging through activation of cognate nuclear receptors (ERa, PR isoforms a and b). In addition to synthesis of PR in female hypothalamic ventromedial nucleus (VMN), E induces changes in local microcircuitry, an effect that requires cAMP activity and for receptivity. Since cAMP increases synthesis and release of constitutive pituitary adenylyl cyclase-activating polypeptide (PACAP) in other tissue for local binding to PACAP receptors, the goal of this proposal is to determine whether E-induced PACAP mediates a novel autocrine/paracrine trans-synaptic loop within VMN microcircuitry required for onset of P-facilitated receptivity. Since studies on the biological role of PR have been hampered by a lack of knowledge of the cell-lineages that express PR isoforms, mice with selective ablation of PRa (PRAKO) and PRb (PRBKO) will be studied. The specific aims of this project are: 1) To determine whether E and P and their cognate receptors [ERa, PRa, PRb] regulate the synthesis and release of pituitary adenylyl cyclase-activating polypeptide (PACAP) in the microcircuitry of the VMN ultimately for receptivity, 2) To characterize the gene expression profile ('molecular fingerprints') of individual PR-expressing and adjacent nonPR- expressing cells in the VMN that mediate isoform-specific PR- facilitated behavior, 3) To identify the steroid-induced mechanism(s) and its major components (VMN expression pattern of membrane-bound receptors [PAC1, VPAC, VPAC2]; signaling pathway [cAMP, MAPK] by which PACAP mediates isoform-specific PR- facilitated behavior. Well-established procedures [in situ hybridization, blot analysis, immunohistochemistry, steroid receptor-dependent behavioral] and new technologies [real-time RT-PCR, single cell expression profiling (molecular fingerprinting), microarrays, Laser capture of individual cells] will be used. By monitoring coordinate changes in PACAP activity, intracellular signaling, and gene expression induced specific steroid receptors (PRa and PRb) in the individual VMN cells that correlate with hormone status and behavior, unique insight into the biological consequences of expressed genetic variability of cells will be identified. This will furnish insight into the underlying cellular and molecular mechanisms that produce steroid receptor-dependent disease etiologies and should contribute to the development of new therapeutic strategies.

卵巢类固醇激素雌激素（E）和孕激素（P）通过激活同源核受体（ER α、PR亚型a和B）对发育、生殖和衰老产生深远影响。 除了在女性下丘脑腹内侧核（VMN）中合成PR外，E还诱导局部微电路的变化，这种作用需要cAMP活性和感受性。 由于cAMP增加的合成和释放的组成型垂体腺苷酸环化酶激活多肽（PACAP）在其他组织中的本地结合到PACAP受体，本提案的目标是确定是否E诱导的PACAP介导一个新的自分泌/旁分泌跨突触环VMN微电路需要启动P-易接受性。 由于对表达PR亚型的细胞谱系缺乏了解而阻碍了对PR生物学作用的研究，因此将研究PRa（PRAKO）和PRb（PRBKO）选择性消融的小鼠。 该项目的具体目标是：1）为了确定E和P及其同源受体[ER a、PRa、PRb]是否调节VMN微回路中垂体腺苷酸环化酶激活多肽（PACAP）的合成和释放以最终获得感受性，2）表征基因表达谱VMN中介导同种型特异性PR促进行为的单个PR表达细胞和相邻的非PR表达细胞的分子指纹（"分子指纹"），3）确定激素诱导的机制及其主要成分（膜结合受体[PAC 1，VPAC，VPAC 2]的VMN表达模式; PACAP介导亚型特异性PR易化行为的信号通路[cAMP，MAPK]。 将使用成熟的程序[原位杂交、印迹分析、免疫组织化学、类固醇受体依赖性行为]和新技术[实时RT-PCR、单细胞表达谱（分子指纹）、微阵列、单个细胞的激光捕获]。 通过监测个体VMN细胞中PACAP活性、细胞内信号传导和基因表达诱导的与激素状态和行为相关的特异性类固醇受体（PRa和PRb）的协调变化，将鉴定对细胞表达的遗传变异性的生物学后果的独特见解。 这将提供深入了解潜在的细胞和分子机制，产生类固醇受体依赖性疾病的病因，并应有助于新的治疗策略的发展。