ULTRASTRUCTURE AND FUNCTION OF NERVE AND MUSCLE

描述：大脑中每个神经元的活动都受到 许多“慢”神经递质的活动不断地从 其他神经元，这些动作的总和决定 神经元的兴奋性。这项拨款提案的主要目标是 阐明“慢”兴奋和慢刺激的生理机制。 “慢”抑制性多肽递质对大脑产生作用 神经元。Meynert核和底核的胆碱能神经元 体外培养蓝斑去甲肾上腺素能神经元 将使用大鼠的大脑；培养这些神经元的技术是 最早于1985年在PI的实验室开发。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，以及 人类的这些神经元在阿尔茨海默病中表现出变性。这个 项目的重点是下列信号转导机制 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 减少阿尔茨海默氏症的发病率。 将特别强调这些发射机如何调制 内向整流钾通道的活性。在许多脑神经元中， 内向整流K通道的调制负责 产生慢兴奋性和慢抑制性突触电位。 具体的项目是：(1)确定哪种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血管活性的影响 K通道；(2)研究第二推拿器的作用 (磷脂酶C和蛋白激酶C，环AMP和环AMP- 信号中的依赖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， 蛋白激酶和具有内向整流性K通道的磷酸酶 通过使用由内而外的贴片。膜片钳电生理学和 将使用免疫细胞化学方法。此外，还将安装一个微型注射器 用于预形成细胞内注射的多肽、抗体和 反义DNA以破坏特定介体的功能 信号转导的级联。

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.