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: 3325232
• 负责人: ROCHELLE S. COHEN
• 金额: $ 5.95万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 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 1991-06-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3325232
• 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.

本提案的主要目标是确定小区 涉及突触和树突变化的生物学机制 伴随雌激素对女性影响的中脑形态 生殖行为，即脊柱前凸。 拟议的研究重点是 腹内侧核（VMN）之间的特定神经元回路 下丘脑和中脑中央灰质（MCG）。雌激素可能会刺激 VMN 中的 RNA 和蛋白质合成机制，导致 多肽的生产，其包装在典型的分泌型中 时尚并沿着 VMN 神经元的轴突运输到 MCG，在那里它可能 作为脊柱前凸通路中的营养因子。拟议的营养级 因子可能诱导突触的生化和形态学改变 MCG 中的树突，这反过来可能会改变突触功效和神经 电路，分别。这个假设的三个方面 已解决。 其中之一是确定 RNA 和蛋白质合成是否 VMN 神经元和 VMN 轴突中的轴突运输是雌激素所必需的 诱导 MCG 突触形态的改变。 这种可能性 将通过注入 RNA 和蛋白质合成抑制剂进行测试或 秋水仙碱，分别进入 VMN 并检查超微结构 随后接受雌激素的动物 MCG 中的突触 治疗。雌激素对潜在营养物质运输的影响 将使用电子显微镜 (EM) 检查催乳素因子 免疫细胞化学。 第二步是检查形态和 雌激素引起的改变的生化机制 MCG 中的突触结构。 对于形态学研究， 雌激素诱导的突触改变序列可能导致 突触数量的增加将通过电镜检查。对于生化 研究发现，磷酸化状态对雌激素的依赖性 MCG 突触中的特定蛋白质将使用亚细胞检查 分级分离技术、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会诱发 与 MCG（和 VMN）树突发育有关的蛋白质。 微管相关蛋白的抗体将用于 EM 免疫细胞化学。形态学和形态学基础研究 明确的哺乳动物行为的生化相关性，例如 脊柱前凸，将有助于理解神经内分泌和 涉及其他复杂行为的细胞生物学机制及其 像差。