ULTRASTRUCTURE AND FUNCTION OF NERVE AND MUSCLE

描述：大脑中每个神经元的活动受到以下因素的影响： 许多“缓慢”的神经递质不断地从 其他神经元，这些动作的总和决定了 神经元的兴奋性。该拨款提案的主要目标是 阐明“缓慢”兴奋和 “缓慢”的抑制肽递质对大脑产生作用 神经元。来自 Meynert 基底核的胆碱能神经元和 培养物中来自蓝斑的去甲肾上腺素能神经元 将使用大鼠大脑；培养这些神经元的技术是 1985 年首次在 PI 实验室开发。这些神经元是 大脑中乙酰胆碱和去甲肾上腺素的主要来源 人类这些神经元在阿尔茨海默病中表现出退化。的 项目重点关注以下信号转导机制 递质：P物质、神经降压素、促肾上腺皮质激素释放因子、 和生长抑素。据报道，其中一些发射器 阿尔茨海默病减少。 将特别强调这些发射机如何调制 内向整流器 K 通道的活动。在许多大脑神经元中， 内向整流器 K 通道的调制负责 缓慢兴奋性和缓慢抑制性突触电位的产生。 具体项目是：（1）鉴定哪种G蛋白介导 P物质、神经降压素、 生长抑素和促肾上腺皮质激素释放因子对活性的影响 K通道； (2)探讨第二按摩器的作用 （磷脂酶 C 和蛋白激酶 C、环 AMP 和环 AMP- 信号中依赖的蛋白激酶和蛋白磷酸酶） 这些递质的转导，以及（2）研究其机制 G 蛋白亚基（突变和非突变）的相互作用， 具有内向整流 K 通道的蛋白激酶和磷酸酶 通过使用由内而外的补丁。膜片钳电生理学和 将使用免疫细胞化学。此外，还将配备显微注射器 用于进行肽、抗体等的细胞内注射 反义 DNA 以破坏特定介质的功能 信号转导级联。

DESCRIPTION: The activity of each neuron in the brain is influenced by the actions of many "slow" neurotransmitters constantly arriving from other neurons, and the total sum of these actions determines the excitability of the neuron. The main objective of this grant proposal is to elucidate the physiological mechanisms by which "slow" excitatory and "slow" inhibitory peptide transmitters produce their actions on brain neurons. Cholinergic neurons from the nucleus basalis of Meynert and noradrenergic neurons from the locus coeruleus in culture prepared from the rat brain will be used; the technique for culturing these neurons was first developed in the PI's laboratory in 1985. These neurons are the main source of acetylcholine and noradrenaline in the brain, and in humans these neurons show degeneration in Alzheimer's disease. The projects focus on the signal transduction mechanisms of the following transmitters: substance P, neurotensin, corticotropin-releasing factor, and somatostatin. Some of these transmitters have been reported to be reduced in Alzheimer's disease. Particular emphasis will be placed on how these transmitters modulate the activity of the inward rectifier K+ channel. In many brain neurons, the modulation of the inward rectifier K+channel is responsible for the generation of slow excitatory and slow inhibitory synaptic potentials. The specific projects are: (1) to identify which G protein mediates the signal transduction of the effects of substance P, neurotensin, somatostatin, and corticotropin-releasing factor on the activity of the K+ channels; (2) to investigate the roles of second massagers (phospholipase C and protein kinase C, cyclic AMP and cyclic AMP- dependent protein kinase, and protein phosphatase) in the signal transduction of these transmitters, and (2) to investigate the mechanisms of the interaction of G protein subunits (mutated and non-mutated), protein kinase, and phosphatases with the inward rectifier K+ channels by using inside-out patches. Patch-clamp electrophysiology and immunocytochemistry will be used. In addition, a microinjector will be used to preform intracellular injection of peptides, antibodies, and antisense DNA in order to disrupt the function of a particular mediator of the signal transduction cascades.