Mechanisms of Ethanol Action on NMDA Receptors and LTP

描述(由申请人提供)：N-甲基D-天冬氨酸受体(NMDAR)是在海马CA1区诱导LTP所需的离子型谷氨酸受体。上世纪80年代，S发现乙醇可抑制这些受体并阻断LTP。乙醇抑制NMDARs和LTP的机制(S)尚未阐明。我们以前已经证明，PKC激活剂诱导NMDAR的酪氨酸磷酸化和增强。LTP的诱导也需要磷酸化。我们推测，乙醇的抑制作用可能是由于NMDAR亚基酪氨酸磷酸化减少导致NMDAR内吞。为了验证这些假设，我们将使用海马片制备、免疫沉淀和Western blotting来评估原位酒精暴露后NMDA亚单位的酪氨酸磷酸化。此外，我们还将利用电生理学研究NMDAR的功能和LTP的诱导。将使用生物化学方法来评估NMDARs的表面表达。我们希望这些研究通过提高我们对乙醇如何影响认知的了解，有助于理解酒精中毒。

DESCRIPTION (provided by applicant): N-methyl D-aspartate receptors (NMDARs) are ionotropic glutamate receptors required for induction of LTP in area CA1 of the hippocampus. In the 1980's, it was discovered that ethanol inhibits these receptors and blocks LTP. The mechanism(s) by which ethanol inhibits NMDARs and LTP have not been elucidated. We have previously shown that PKC activators induce tyrosine phosphorylation and potentiation of the NMDAR. Phosphorylation is also required for LTP induction. We hypothesized that the inhibitory action of ethanol may be a result of reduced tyrosine phosphorylation of NMDAR subunits leading to endocytosis of NMDARs. In order to test these hypotheses, we will use the hippocampal slice preparation, immunoprecipitation, and western blotting to evaluate tyrosine phosphorylation of NMDA subunits following in situ ethanol exposure. Additionally, we will study NMDAR function and LTP induction utilizing electrophysiology. Biochemical methods will be employed to evaluate the surface expression of NMDARs. We hope that these studies will contribute to the understanding of alcoholism by enhancing our knowledge of how ethanol affects cognition.