CELL BIOLOGY OF ESTROGEN EFFECTS ON LORDOSIS BEHAVIOR

雌激素对脊柱行为影响的细胞生物学

• 批准号: 3325229
• 负责人: ROCHELLE S. COHEN
• 金额: $ 7.33万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-09-01 至 1994-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3325229
• 关键词: RNA; autoradiography; cell biology; dendrites; electron microscopy; estrogens; female; hypothalamus; immunocytochemistry; laboratory rat; mesencephalon; microtubule associated protein; neural conduction; neural transmission; neuroanatomy; neurobiology; neurogenesis; neuronal transport; periaqueductal gray matter; phosphorylation; prolactin; protein biosynthesis; sex behavior; synapses

The main objective of the present proposal is to determine the cell biological mechanisms involved in changes in synaptic and dendritic morphology in the midbrain that accompany estrogen effects on female reproductive behavior, i.e., lordosis. The proposed studies focus on the specific neuronal circuit between the ventromedial nucleus (VMN) of the hypothalamus and the midbrain central gray (MCG). Estrogen may stimulate RNA and protein synthetic mechanisms in the VMN, resulting in the production of a polypeptide, which is packaged in a typical secretory fashion and transported down axons of VMN neurons to the MCG, where it may act as a trophic factor in the pathway for lordosis. The proposed trophic factor may induce biochemical and morphological alterations in synapses and dendrites in the MCG, which in turn may alter synaptic efficacy and neural circuitry, respectively. Three aspects of this hypothesis will be addressed. One will be to determine whether RNA and protein synthesis in VMN neurons and axonal transport in VMN axons are required for estrogen- induced alterations in synaptic morphology in the MCG. This possibility will be tested by infusing inhibitors of RNA and protein synthesis or colchicine, respectively, into the VMN and examining the ultrastructure of synapses in the MCG of animals which have received subsequent estrogen treatment. The effect of estrogen on the transport of the potential trophic factor, prolactin, will be examined using electron microscopic (EM) immunocytochemistry. The second will be to examine the morphological and biochemical mechanisms underlying the estrogen-induced alterations in synaptic structures in the MCG. For the morphological studies, the sequence of estrogen-induced synaptic alterations which may lead to an increase in synaptic number will the examined by EM. For the biochemical studies, the dependence on estrogen of the phosphorylation state of specific proteins in MCG synapses will be examined using subcellular fractionation techniques, in vitro phosphorylation, and SDS-gel autoradiography. The third way will be to determine if estrogen induces a protein implicated in dendritic development in the MCG (and VMN). Antibodies to microtubule-associated proteins will be used for EM immunocytochemistry. Fundamental studies on the morphological and biochemical correlates of a well-defined mammalian behavior, such as lordosis, will contribute to an understanding of the neuroendocrine and cell biological mechanisms involved in other complex behaviors and their aberrations.

本提案的主要目的是确定细胞 突触和树突变化涉及的生物学机制 中脑的形态伴随雌激素对女性的影响 生殖行为，即洛里森。 拟议的研究着重于 特定的神经元电路之间的腹侧核（VMN）之间 下丘脑和中脑中央灰色（MCG）。雌激素可能会刺激 VMN中的RNA和蛋白质合成机制，导致 产生多肽，该多肽是在典型分泌中包装的 时尚，并将VMN神经元的轴突运送到MCG，可能 充当脊柱障碍途径的营养因素。提出的营养 因素可能诱导突触的生化和形态改变， MCG中的树突又可能会改变突触功效和神经 电路分别。该假设的三个方面将是 解决。 一种将是确定RNA和蛋白质合成是否在 VMN神经元和VMN轴突中的轴突转运是雌激素需要的 MCG中突触形态的诱导改变。 这种可能性 将通过注入RNA和蛋白质合成抑制剂或 秋水仙碱分别进入VMN并检查的超微结构 在接收随后的雌激素的动物MCG中的突触 治疗。雌激素对电势营养的运输的影响 因子（催乳素）将使用电子显微镜（EM）检查 免疫细胞化学。 第二个是检查形态学和 雌激素诱导改变的生化机制 MCG中的突触结构。 对于形态学研究， 雌激素诱导的突触改变的序列，这可能导致 突触数的增加将由EM检查。对于生化 研究，依赖对磷酸化状态的雌激素的依赖性 MCG突触中的特定蛋白质将使用亚细胞检查 分馏技术，体外磷酸化和SDS-凝胶 放射自显影。 第三种方法是确定雌激素是否诱导A 蛋白质与MCG（和VMN）中的树突状发育有关。 微管相关蛋白的抗体将用于EM 免疫细胞化学。关于形态学和 定义明确的哺乳动物行为的生化相关性，例如 dordosis，将有助于理解神经内分泌和 细胞生物学机制涉及其他复杂行为及其 畸变。