MODERATE ETHANOL PRENATALLY ON NMDA PHOSPHORYLATION

产前适度乙醇对 NMDA 磷酸化

• 批准号: 6509279
• 负责人: STEVEN W LESLIE
• 金额: $ 28.6万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6509279
• 关键词: NMDA receptors; RNase protection assay; SDS polyacrylamide gel electrophoresis; brain metabolism; cerebellum; cerebral cortex; dosage; embryo /fetus toxicology; ethanol; fetal alcohol syndrome; glutamates; hippocampus; immunoprecipitation; laboratory rat; messenger RNA; newborn animals; phosphorylation; protein kinase; protein structure function; receptor binding; receptor expression; vertebrate embryology; western blottings

Studies in our laboratory and by others indicate that prenatal and early postnatal ethanol exposure results in a significant reduction of NMDA receptor function and number. NMDA receptors are linked with important aspects of brain development and learning and memory. Ethanol-related deficits in NMDA function, therefore, may be of key importance not only in the severe cognitive and behavioral deficiencies of fetal alcohol syndrome (FAS) but also in alcohol-related neurodevelopmental disorders (ARND) typically expressed as behavioral symptoms associated with fetal alcohol effects (FAE). We have recently found that decreases in NMDA receptor function resulting from prenatal ethanol exposure in rats are accompanied by reductions in some, but not all, of the subunits comprising the NMDA receptor complex. NMDAR2A and NMDAR2B subunits are significantly reduced, while the NMDAR1 subunit is unaffected. Studies conducted thus far have examined the effects of high doses of ethanol prenatally and postnatally on NMDA receptor function and subunit levels. Studies proposed in specific aims 1 through 4 are designed to determine the dose-response relationships of prenatal ethanol treatment on 1) functional measures of NMDA receptor activation using fura-2 loaded dissociated neurons and H-MK801 binding, 2) NMDAR1 and NMDAR2 subunit protein (Western blot analysis) and messenger RNA levels (RNase protection assay), and 3) changes in NMDA receptor subunit composition as measured by immunoprecipitation studies. Specific aim 4 will examine the relationship(s) of NMDA receptor subunit changes with changes in subunit phosphorylation. The overall hypothesis of this proposal is that prenatal ethanol exposure, even at modest concentrations, will result in demonstrable deficits in the function of NMDA receptors. We expect the magnitude of the deficits to be dose-related. It is hypothesized further that ethanol's actions in this regard may be linked with abnormalities of NMDA receptor phosphorylation.

在我们的实验室和其他研究中的研究表明，产前和早期产后乙醇暴露会显着降低NMDA受体功能和数量。 NMDA受体与大脑发育，学习和记忆的重要方面有关。 因此，与乙醇相关的NMDA功能缺陷，不仅在胎儿酒精综合征（FAS）的严重认知和行为缺陷中，而且在与酒精相关的神经发育障碍（ARND）中，不仅在与胎儿酒精效应相关的行为症状（FAE）相关的行为症状（FAE）。 我们最近发现，大鼠产前乙醇暴露引起的NMDA受体功能的降低伴随着包括NMDA受体复合物的一些亚基的降低但不是全部。 NMDAR2A和NMDAR2B亚基显着降低，而NMDAR1亚基不受影响。 迄今为止进行的研究已经检查了产前和产后乙醇对NMDA受体功能和亚基水平的影响。在特定目的1到4中提出的研究旨在确定产前乙醇处理在1）使用Fura-2加载的解离神经元和H-MK801结合的NMDA受体激活的功能测量的剂量反应关系，2）NMDAR1和NMDAR1和NMDAR2亚基蛋白（Western blot rantase sance）和3级均变化（NMDAR）（nmdar2通过免疫沉淀研究测量的受体亚基组成。 具体目标4将检查NMDA受体亚基的关系随着亚基磷酸化的变化而变化。 该提议的总体假设是，即使以适度的浓度，产前乙醇的暴露也会导致NMDA受体功能的明显缺陷。 我们期望缺陷的大小与剂量有关。 进一步假设，这方面的乙醇的作用可能与NMDA受体磷酸化的异常有关。