Analysis of molecular mechanisms for localization and functional modulation of metabotropic glutamate receptors in the central nervous system

中枢神经系统代谢型谷氨酸受体定位和功能调节的分子机制分析

• 批准号: 14570106
• 负责人: NAKAJIMA Yoshiaki
• 金额: $ 2.56万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570106/
• 关键词: glutamate receptor; cerebellum; reversible; Cajal-Retzius cells; striatum; acetylcholine; tamalin; scaffold protein; グルタミン酸; シナプス

In the cerebellar circuit, we studied the role and synaptic mechanisms of postsynaptic metabotropic glutamate receptor subtype 2 (mGluR2) at granule cell-Golgi cell synapses, using whole-cell recording of green fluorescent protein-positive Golgi cells of wild-type and mGluR2-deficient mice. We found that postsynaptic mGluR2 thus senses inputs from granule cells and is most likely important for spatiotemporal modulation of mossy fiber-granule cell transmission before distributing inputs to Purkinje cells.The striatum in the basal ganglia-thalamocortical circuitry is a key neural substrate that is implicated in motor balance and procedural learning. The projection neurons in the striatum are dynamically modulated by nigrostriatal dopaminergic input and intrastriatal cholinergic input. The role of intrastriatal acetylcholine (ACh) in learning behaviors, however, remains to be fully clarified. In this investigation, we examine the involvement of intrastriatal ACh in different categories of … More learning by selectively ablating the striatal cholinergic neurons with use of immunotoxin-mediated cell targeting. We found that cholinergic modulation in the local striatal circuit plays a pivotal role in regulation of neural circuitry Involving reward-related procedural learning and working memory.We developed a novel technique that allowed reversible suppression of glutamatergic neurotransmission in the cerebellar network. The transgenic mice will serve as an animal model to study the cerebellar function in motor coordination and learning.Layer 1 In the developing cerebral cortex is populated by two basic neuronal cell types. Cajal-Retzius (CR) cells and non-CR cells. We generated transgenic mice in which green fluorescent protein (GFP) was driven by the promoter of metabotropic glutamate receptor subtype 2 and expressed specifically in CR cells during cortical development. The resuts imply that the layer 1 neurons dynamically change and play a distinct and integral role in the postnatal developing neocortex.Tamalin is a scaffold protein that comprises multiple protein-interacting domains, including a 95-kDa postsynaptic density protein (PSD-95)/discs-large/ZQ-1 (PDZ) domain, a leucine-zipper region, and a carboxyl-terminal PDZ binding motif. Tamalin forms a complex with metabotropic glutamate receptors and guanine nucleotide exchange factor cytohesins and promotes intracellular trafficking and cell surface expression of group 1 metabotropic glutamate receptors. In the present study, using several different approaches we have shown that tamalin interacts with multiple neuronal proteins through Its distinct protein-binding domains. The results indicate that tamalin exists as a key element that forms a protein complex with multiple postsynaptic and protein-trafficking scaffold proteins. Less

在小脑回路中，我们利用野生型和mGluR2缺陷小鼠绿色荧光蛋白阳性高尔基细胞的全细胞记录，研究了突触后代谢型谷氨酸受体亚型2 （mGluR2）在颗粒细胞-高尔基细胞突触中的作用和突触机制。我们发现突触后mGluR2感知来自颗粒细胞的输入，并且在将输入分配给浦肯野细胞之前，很可能对苔藓纤维-颗粒细胞传输的时空调节很重要。基底神经节-丘脑皮层回路中的纹状体是参与运动平衡和程序性学习的关键神经基质。纹状体投射神经元受黑质纹状体多巴胺能输入和纹状体内胆碱能输入的动态调节。然而，胃内乙酰胆碱（ACh）在学习行为中的作用仍有待充分阐明。在这项研究中，我们通过使用免疫毒素介导的细胞靶向选择性地切除纹状体胆碱能神经元，研究了纹状体内乙酰胆碱能参与不同类别的学习。我们发现，局部纹状体回路的胆碱能调节在与奖励相关的程序学习和工作记忆的神经回路中起着关键作用。我们开发了一种新的技术，可以可逆地抑制小脑网络中的谷氨酸能神经传递。该转基因小鼠将作为研究小脑运动协调和学习功能的动物模型。第一层发育中的大脑皮层由两种基本的神经元细胞组成。Cajal-Retzius细胞和非CR细胞。我们培育了绿色荧光蛋白（GFP）由代谢性谷氨酸受体亚型2启动子驱动的转基因小鼠，并在皮质发育期间在CR细胞中特异性表达。结果表明，第1层神经元在出生后大脑皮层的发育中起着独特而完整的作用。Tamalin是一种支架蛋白，由多个蛋白相互作用结构域组成，包括95 kda突触后密度蛋白(PSD-95)/disc -large/ZQ-1 （PDZ）结构域、亮氨酸拉链区和羧基末端PDZ结合基元。他马林与代谢性谷氨酸受体和鸟嘌呤核苷酸交换因子胞苷形成复合物，促进细胞内转运和1组代谢性谷氨酸受体的细胞表面表达。在目前的研究中，使用几种不同的方法，我们已经表明tamalin通过其不同的蛋白质结合域与多种神经元蛋白相互作用。结果表明，tamalin是与多个突触后和蛋白质转运支架蛋白形成蛋白质复合物的关键元素。少

项目成果

Kitabatake, Y.: "Impairment of reward-related learning by cholinergic cell ablation in the striatum"Proc.Natl.Acad.Sci.USA. 93. 13515-13522 (2003)

Kitabatake，Y.：“纹状体胆碱能细胞消融对奖励相关学习的损害”Proc.Natl.Acad.Sci.USA。

Dai Watanabe, Shigetada Nakanishi: "mGluR2 Postsynaptically Senses Granule Cell Inputs at Golgi Cell Synapses"Neuron. Vol.39. 821-829 (2003)

Dai Watanabe、Shigetada Nakanishi：“mGluR2 突触后感知高尔基细胞突触的颗粒细胞输入”神经元。

Hazama, M., Watanabe, D., Suzuki M., Mizoguchi, A., Pastan, I., Nakanishi, S.: "Different regulatory sequences are required for parvalbumin gene expression in skeletal muscles and neuronal cells of transgenic mice"Brain Res Mol Brain Res. 110. 53-66 (2002

Hazama, M.、Watanabe, D.、Suzuki M.、Mizoguchi, A.、Pastan, I.、Nakanishi, S.：“转基因小鼠骨骼肌和神经元细胞中的小清蛋白基因表达需要不同的调控序列”

Yasuji Kitabatake, Takatoshi Hikida, Dai Watanabe, Ira Pastan, Shigetada Nakanishi: "Impairment of reward-related learning by cholinergic cell ablation in the striatum"PNAS. Vol.100. 7965-7970 (2003)

Yasuji Kitabatake、Takatoshi Hikida、Dai Watanabe、Ira Pastan、Shigetada Nakanishi：“纹状体胆碱能细胞消融对奖励相关学习的损害”PNAS。

Takeshi Soda, Ryo Nakashima, Dai Watanabe, Kazunori Nakajima, Ira Pastan, Shigetada Nakanishi: "Segregation and Coactivation of Developing Neocortical Layer 1 Neurons"J.Neurosci.. Vol.23. 6272-6279 (2003)

Takeshi Soda、Ryo Nakashima、Dai Watanabe、Kazunori Nakajima、Ira Pastan、Shigetada Nakanishi：“发育中新皮质第 1 层神经元的分离和共激活”J.Neurosci.. Vol.23。