Gene Expression in Nervous Tissue

神经组织中的基因表达

• 批准号: 7100547
• 负责人: Bruce S. McEwen
• 金额: $ 39.26万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1974
• 资助国家: 美国
• 起止时间: 1974-09-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7100547
• 关键词: electron microscopy; estrogen receptors; estrogens; gene expression; gene targeting; genetically modified animals; glia; hippocampus; hormone regulation /control mechanism; immunocytochemistry; in situ hybridization; interneurons; laboratory mouse; light microscopy; microarray technology; neurogenetics; northern blottings; phosphorylation; polymerase chain reaction; progesterone; progestins; protein biosynthesis; receptor expression; second messengers; synaptogenesis

DESCRIPTION (provided by applicant): The ovarian hormones estradiol (E) and progesterone (P) have important effects on higher brain function besides their role in reproduction, and they affect many brain regions besides the hypothalamus, including the hippocampus and cerebral cortex. This project addresses molecular mechanisms by which E and P regulate spine synapse formation and maturation (SSF/M) in the hippocampus, a brain region involved in learning and memory of events in daily life. Synapse turnover occurs cyclically during the rat ovarian cycle by a mechanism that requires the activity of NMDA receptors and the participation of genomic and non-genomic estrogen (ER) and progestin receptors (PR). E-dependent SSF/M occurs in hippocampus and in prefrontal cortex in rhesus monkeys, as well as in rats and mice. There is electrophysiological, neuroanatomical and behavioral detail for this phenomenon, which has established its potential relevance for effects of E deprivation on human cognitive function. The two forms of the E receptor, ERa and ER¿, as well as PR are found in both nuclear and non-nuclear sites in hippocampal neurons, and emerging evidence indicate that E and P affect non-genomic signaling pathways. Our central hypothesis is that E and P effects upon SSF/M and removal involve rapid non-genomic hormone actions, as well as direct and indirect genomic effects, leading to the modification of the actin cytoskeleton, as well as the stimulation of protein synthesis and coordinated genomic activity that leads to SSF/M. We shall address the role of ER subtypes in SSF/M and memory using specific agonists for each receptor isoform. Because mice are useful for genetic manipulations, we shall study key aspects of E and P action on SSF/M in mice. Studies will be carried out on mice lacking ERa and ER¿ as well as on mice with an inhibitory form of the transcription regulator, CREB, that may normally mediate indirect genomic actions of E. We shall determine the role of E dependent signaling pathways in SSF/M based upon data we have obtained from in vitro studies of E-responsive NG108-15 cells in order to study hippocampal neurons in vitro using confocal microscopy and then in vivo using light and electron microscopy. We shall study non-nuclear signaling actions of E, namely, the regulation of phosphorylation of Akt and the phosphorylation of LIMK-1 leading to cofilin phosphorylation and actin polymerization. Finally, since E-regulated spine SSF/M is reversed rapidly by P, we shall investigate how P causes loss of spines via the ubiquitin-proteasome pathway. These studies will provide new basic information to better understand and treat cognitive and other higher brain functions that are affected by surgical- or age-related loss of ovarian hormones during menopause, as well as protection of the brain from seizures and stroke by E and P.

描述（由适用提供）：除了在繁殖中的作用外，卵巢恐怖雌激素雌二醇（E）和孕酮（P）对较高的大脑功能具有重要作用，并且除了下丘脑外，还影响了许多大脑区域，包括海马和脑皮质。该项目探讨了E和P调节海马中脊柱突触形成和成熟（SSF/M）的分子机制，海马是一个涉及日常生活中事件的大脑区域。突触转换在大鼠卵巢周期期间通过需要NMDA受体活性以及基因组和非基因组雌激素（ER）和孕激素受体（PR）的参与的机制进行周期性发生。 E依赖性SSF/M发生在海马和恒河猴以及大鼠和小鼠中的前额叶皮层中。这种现象有电生理，神经解剖学和行为细节，该现象已经建立了其对E剥夺对人类认知功能的影响的潜在相关性。 E受体，ERA和ER？的两种形式以及PR在海马神经元中的核和非核部位都发现，以及新出现的证据，表明E和P会影响非基因组信号传导途径。我们的中心假设是E和P对SSF/M的影响并消除了快速的非基因组恐怖措施作用以及直接和间接的基因组效应，从而导致肌动蛋白细胞骨架的修饰，以及刺激蛋白质合成和均衡的基因组活性，从而导致SSF/M。我们应使用每个受体同工型使用特定的激动剂来解决ER亚型在SSF/M中的作用。由于小鼠对遗传操作很有用，因此我们将研究小鼠SSF/M的E和P作用的关键方面。将对缺乏ERA和ER的小鼠以及具有转录调节剂CREB的抑制作用形式的小鼠进行研究，该抑制作用通常可能是培养基。我们应根据数据在SSF/M中的E依赖信号途径的作用，基于SSF/M的依赖性信号途径的作用，基于数据，我们已经通过e-e-respons ng108-15 ng108-15细胞研究的数据中的数据来确定依赖性信号的作用。显微镜，然后使用光和电子显微镜在体内。我们将研究E的非核信号传导作用，即，Akt的磷酸化调节以及limk-1的磷酸化导致Cofilin磷酸化和肌动蛋白聚合的磷酸化。最后，由于p迅速将E调节的脊柱SSF/M迅速逆转，因此我们将研究P如何通过泛素 - 蛋白酶体途径导致脊柱丧失。这些研究将提供新的基本信息，以更好地理解和治疗受外科手术或年龄相关的卵巢骑兵损失的认知和其他较高的大脑功能，并保护大脑免受癫痫发作的保护，并受到E和P的中风。