NEUROENDOCRINE BASES OF REPRODUCTIVE BEHAVIOR

生殖行为的神经内分泌基础

• 批准号: 2485769
• 负责人: ANNE M ETGEN
• 金额: $ 29.17万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2485769
• 关键词: alpha adrenergic receptor; arrestins; behavioral /social science research tag; beta adrenergic receptor; biological signal transduction; drug administration rate /duration; enzyme activity; estradiol; female; guanylate cyclase; hormone regulation /control mechanism; hypothalamus; immunocytochemistry; laboratory rat; luteinizing hormone; messenger RNA; neuroendocrine system; nitric oxide; norepinephrine; phospholipase C; preoptic areas; progesterone; receptor expression; sex behavior; western blottings

(Adapted from the investigator's abstract): The ovarian hormones estradiol (E) and progesterone (P) act in the hypothalamus and preoptic area (HPOA) to stimulate the preovulatory release of pituitary gonadotropins and coordinate the expression of reproductive mating behavior, namely lordosis by the female through the action of norepinephrine (NE). The goal of the proposed research is to examine the molecular mechanisms by which estradiol and progesterone modulate signal transduction on alpha1-and beta-adrenoceptors in the HPOA and to relate these to the expression of reproductive behavior. Specific Aim 1 will test the hypothesis that estradiol elevates alpha1B-adrenoceptors in populations of HPOA neurons that express ER. Immunocytochemical approaches will be employed to determine: 1) whether E increases alpha1B-adrenoceptor protein in regions of the HPOA that also express ER; 2) whether A1B - adrenoceptors and ER are colocalized in some or all of these neurons; and 3) whether hypothalamic neurons expressing alpha1B-adrenoceptors project to the midbrain central gray (MCG). Specific aim 2 will test the hypothesis that in the HPOA of E-primed females P will switch alpha1 adrenoceptor signaling from activation of phospholipase C to calcium- dependent activation of the nitric oxide (NO)/soluble guanylyl cyclase pathway. Support for this hypothesis is derived from the observation that alpha1-adrenergic activation of NO synthesis influences the preovulatory release of LH and because expression of reproductive behavior in E+P treated female rats is inhibited by NO synthetase inhibitors and inhibitors of soluble guanylyl cyclase. Specific aim 3 will test the hypothesis that E increases the expression of one or more protein kinase C (PKC) isoenzymes in the HPOA. PKC is a major downstream mediator of alpha1-adrenergic signal transduction; hence, induction of PKC could further amplify alpha1-adrenergic signaling in the HPOA. Experiments will utilize assays of PKC catalytic activity as well as phorbol ester binding and Western Blots to identify the separate isoenzymes. Specific Aim 4 will test the hypothesis that E regulates molecules involved in adrenergic receptor-G protein coupling. Molecular biological and immunological methods will be used to determine the effects of E on: 1) mRNA and protein levels of b-adrenergic receptor kinase 1 and 2 (b-ARK1, and b-ARK2) and 2) the mRNA and protein levels of b-arrestin1 and b-arrestin2. These are significant questions because b-ARKs and b-arrestins impede the interactions of b-adrenergic, a2-adrenergic and u-opioid receptors with G proteins, and they find that E treatment decreases the function of all three of these receptors in the HPOA with measurably downregulating the receptors.

（改编自调查员的摘要）：卵巢激素 雌二醇（E）和孕酮（P）作用于下丘脑和前丘脑前的作用 面积（HPOA）刺激垂体的排卵前释放 促性腺激素并协调生殖交配的表达 行为，即女性通过的作用 去甲肾上腺素（NE）。拟议研究的目的是检查 雌二醇和孕酮调节信号的分子机制 HPOA中的α1和β-肾上腺素受体的转导并与之相关 这些表达生殖行为。具体目标1将测试 雌二醇升高α1b-肾上腺素受体的假设 表达ER的HPOA神经元的种群。免疫细胞化学方法 将采用以下确定：1）e是否增加了alpha1b-adrenoceptor HPOA区域的蛋白质也表达ER； 2）A1b-是否 在某些或所有这些神经元中，肾上腺受体和ER共定位。和 3）下丘脑神经元是否表达α1b-adrenoceptors项目 到中脑中央灰色（MCG）。具体目标2将测试 假设在E-apred雌性的HPOA中会切换alpha1 从磷脂酶C激活到钙 - 的肾上腺受体信号传导 一氧化氮（NO）/可溶性鸟类环酶的依赖激活 路径。对这一假设的支持来自于观察到 NO合成的α1-肾上腺素能激活会影响卵巢治疗 LH的释放和E+P中生殖行为的表达 没有合成酶抑制剂抑制治疗的雌性大鼠， 可溶性鸟叶基环酶的抑制剂。特定目标3将测试 E增加了一种或多种蛋白激酶的表达的假设 HPOA中的C（PKC）同工酶。 PKC是主要的下游调解人 α1-肾上腺素能信号转导；因此，PKC的诱导可能 进一步扩增HPOA中的α1-肾上腺素能信号传导。实验会 利用PKC催化活性以及凤凰酯结合的测定 和蛋白质印迹以识别单独的同工酶。具体目标4将 检验E调节肾上腺素能的分子的假设 受体-G蛋白偶联。分子生物学和免疫学 方法将用于确定E ON的影响：1）mRNA和蛋白质 B-肾上腺素能受体激酶1和2（B-ARK1和B-ARK2）和2的水平 B-arrestin1和B-arrestin2的mRNA和蛋白质水平。这些都是 重大问题是因为b-arks和b-arrestin都阻碍了 B-肾上腺素能，A2-肾上腺素能和U-阿片类受体与G的相互作用 蛋白质，他们发现E治疗降低了所有的功能 HPOA中的三个受体，可测量地下调 受体。