MOLECULAR AND CELLULAR NEUROPHYSIOLOGY FOR MENTAL HEALTH

心理健康的分子和细胞神经生理学

• 批准号: 6530873
• 负责人: SHIGEHIRO NAKAJIMA
• 金额: $ 18.14万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-01 至 2004-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6530873
• 关键词: G protein; biological signal transduction; brain cell; calcium channel; cell line; dopamine; electrophysiology; fluorescence microscopy; human tissue; immunocytochemistry; laboratory rat; membrane channels; mental disorders; mental health; molecular pathology; neural inhibition; neurophysiology; neuroregulation; neurotensin; potassium channel; site directed mutagenesis; substantia nigra; tegmentum; tissue /cell culture; transfection; voltage /patch clamp

The objective of this research proposal is to study the regulatory mechanisms of brain neurons that are implicated in major mental disorders. The proposed experiments will be done mainly by using primary cultured dopaminergic (DA) neurons from the substantia nigra and the ventral tegmental area of the rat brain. The methods will be electrophysiological techniques combined with molecular biological methods. The first project is concerned with excitation of these DA neurons. Neurotensin produces a slow excitation of DA neurons by activating a non-selective cation channel. The analysis of the ionic mechanism has revealed that this non-selective channel is different from the well-known non-selective ion channels such as the CAN channels (the calcium-activated non-specific cation channels) or the cyclic nucleotide-gated channels. The proposed experiments will shed light on the signal transduction mechanisms of the neurotensin-induced activation of this non-selective channel. The role played by G proteins will be critically evaluated, and the identity of the G protein will be determine. The other project deals with a slow inhibition of the DA neurons by dopamine itself. This self-inhibition results from an activation of an inward rectifier K+ channel by the G protein betagamma- subunits. The project will investigate the mechanisms of the interaction of the betagamma-subunits and the inward rectifier molecule. It is proposed to find a convenient way to transfect primary cultured DA neurons with plasmid cDNA. Then the DA neurons (also HEK293 cells) will be transfected with CDNAs encoding the G protein beta-subunit and the gamma-subunit. The effects of site-directed mutations of the beta- subunit will be evaluated. The investigation will show which regions of the beta-subunit molecule are responsible for the association with and activation of the inward rectifier molecule.

本研究计划的目的是研究监管 大脑神经元的机制与主要心理有关 失调。 所提出的实验将主要通过使用 原代培养的来自黑质的多巴胺能 (DA) 神经元和 大鼠大脑的腹侧被盖区。 方法将是 电生理技术与分子生物学相结合 方法。 第一个项目涉及这些 DA 的激发 神经元。 神经降压素通过以下方式产生 DA 神经元的缓慢兴奋 激活非选择性阳离子通道。 离子分析 机制表明，这种非选择性通道不同于 众所周知的非选择性离子通道，例如 CAN 通道（ 钙激活的非特异性阳离子通道）或循环 核苷酸门控通道。 拟议的实验将揭示 神经降压素诱导激活的信号转导机制 这个非选择性通道。 G蛋白所起的作用是 严格评估，G 蛋白的身份将被确定 决定。 另一个项目涉及 DA 的缓慢抑制 多巴胺本身的神经元。 这种自我抑制的结果是 G 蛋白 betaamma- 激活内向整流 K+ 通道 亚单位。 该项目将研究该机制 βγ亚基和内向整流分子的相互作用。 建议寻找一种便捷的方法来转染原代培养物 带有质粒 cDNA 的 DA 神经元。 然后是 DA 神经元（也是 HEK293 细胞） 将被编码G蛋白β亚基的cDNA转染并且 伽马亚基。 β-定点突变的影响 将评估子单元。 调查将显示哪些地区 β-亚基分子的负责与 以及内向整流分子的激活。

项目成果

Importance of the G protein gamma subunit in activating G protein-coupled inward rectifier K(+) channels.

G 蛋白 γ 亚基在激活 G 蛋白偶联内向整流 K( ) 通道中的重要性。

• DOI: 10.1016/s0014-5793(99)01656-7
• 发表时间: 1999
• 期刊: FEBS letters
• 影响因子: 3.5
• 作者: Kawano,T;Chen,L;Watanabe,SY;Yamauchi,J;Kaziro,Y;Nakajima,Y;Nakajima,S;Itoh,H
• 通讯作者: Itoh,H

Morphological and physiological properties of serotonergic neurons in dissociated cultures from the postnatal rat dorsal raphe nucleus.

产后大鼠中缝背核分离培养物中血清素能神经元的形态和生理特性。

• DOI: 10.1016/j.jneumeth.2007.08.018
• 发表时间: 2008
• 期刊: Journal of neuroscience methods
• 影响因子: 3
• 作者: Yasufuku-Takano,Junko;Nakajima,Shigehiro;Nakajima,Yasuko
• 通讯作者: Nakajima,Yasuko