NEUROTRANSMITTER REGULATION OF NFG IN THE CNS

这项提案的目标将集中在参与具体的 神经生长因子基因调控中的神经递质系统 在中枢神经系统(CNS)中表达。这些研究描述了 旨在测试神经递质谷氨酸， 通过激活N-甲基-D-天冬氨酸(NMDA)受体和去甲肾上腺素 (NE)参与NGF基因表达的调节。这些 选择神经递质系统的原因有很多。两个都是 众所周知，这些神经递质系统在突触中发挥作用。 可塑性。此外，还有电生理学，神经解剖学， 以及支持这两者相互作用的药理学数据 神经递质系统。我们将分别关注每个系统，并 然后提出假设，这两个神经递质系统 在海马结构和皮质中相互作用以调节 NGF基因的表达。可以推测，这两个因素的激活 神经递质系统可能最终影响功能(生存 和可塑性)神经生长因子反应性胆碱能基底前脑神经元 通过激活NGF的表达。然而，在这个时候，我们将 将我们的努力集中在NGF基因的表达上，了解到 未来的目标是测试去甲肾上腺素和谷氨酸的后果 NMDA诱导的NGF对胆碱能功能的影响。中的研究 这一建议对于进一步理解 CNS中的NGF以及参与其中的潜在过程 与阿尔茨海默病相关的神经退行性变。有证据表明 对于胆碱能、去甲肾上腺素和谷氨酸的功能改变 阿尔茨海默病患者的神经递质系统。 因为基底前脑胆碱能神经元被认为是 对神经生长因子的神经营养特性敏感，研究其调节 NE和谷氨酸对这种神经营养因子的作用可能在 了解一些与以下相关的神经退行性变过程 阿尔茨海默氏症。我们将使用原位杂交组织化学 以及核糖核酸酶保护试验来研究神经递质诱导的 体外(组织切片)和体内NGF mRNA水平的变化。这些 将使用两种技术来量化和本地化 神经生长因子基因在海马区和皮质的表达变化。 在提案的早期阶段，我们将使用 对海马区和皮质组织的具体情况进行调查 神经递质剂量，时间进程，受体药理学，继发性 信使等)了解神经递质调节所必需的 神经生长因子基因的表达。一旦这些条件建立起来，我们将 然后进行体内研究，以定位NGF mRNA的表达 对神经递质挑战有反应的细胞。最后，我们 建议研究这些物质的生理作用(和相互作用) 体内的两种神经递质使用经过充分研究的范例设计成 分别刺激每种神经递质的内源性作用 并行的。

The goals of this proposal will focus on the involvement of specific neurotransmitter systems in regulating nerve growth factor (NGF) gene expression in the central nervous system (CNS). The studies described are designed to test the hypothesis that the neurotransmitters glutamate, via N-methyl-d-aspartate (NMDA) receptor activation, and norepinephrine (NE) are involved in the regulation of NGF gene expression. These neurotransmitter systems were chosen for a number of reasons. Both of these neurotransmitter systems are known to play a role in synaptic plasticity. In addition, there is electrophysiological, neuroanatomical, and pharmacological data supporting the interaction of these two neurotransmitter systems. We will focus on each system separately and then formulate the hypothesis that these two neurotransmitter systems interact in the hippocampal formation and cortex to regulate levels of NGF mRNA. It could be postulated that the activation of these two neurotransmitter systems may ultimately influence the function (survival and plasticity) of NGF responsive cholinergic basal forebrain neurons through activation of NGF expression. However, at this time we will focus our efforts on NGF gene expression, with the understanding that future goals will be to test the consequence of NE and glutamate (NMDA)-induced changes in NGF on cholinergic function. The studies in this proposal are relevant for further understanding of the actions of NGF in the CNS, as well as potential processes involved in neurodegeneration associated with Alzheimer's disease. There is evidence for altered function in the cholinergic, noradrenergic, and glutamate neurotransmitter systems in patients suffering from Alzheimer's disease. Because the basal forebrain cholinergic neurons are thought to be sensitive to the neurotrophic properties of NGF, studying the regulation of this neurotrophic factor by NE and glutamate may be critical in understanding some of the neurodegenerative processes associated with Alzheimer's disease. We will use in situ hybridization histochemistry and a ribonuclease protection assay to study neurotransmitter-induced changes in NGF mRNA levels in vitro (tissue slices) and in vivo. These two techniques will be utilized for quantification and localization of changes in NGF gene expression in the hippocampal formation and cortex. In the early phases of the proposal, we will use tissue slices of hippocampal and cortical tissue to investigate the specific conditions (neurotransmitter dose, time course, receptor pharmacology, secondary messengers, etc.) necessary for understanding neurotransmitter regulation of NGF gene expression. Once these conditions are established, we will then move to in vivo studies in order to localize NGF mRNA expressing cells that are responding to neurotransmitter challenge. Finally, we propose to study the physiological actions (and interactions) of these two neurotransmitters in vivo using well-studied paradigms designed to stimulate the endogenous actions of each neurotransmitter separately and in parallel.