GLUTAMATE TRANSPORTER EAAT4 IN PURKINJE CELL BIOLOGY

浦金野细胞生物学中的谷氨酸转运蛋白 EAAT4

基本信息

批准号：
6027039
负责人：
NICHOLAS J MARAGAKIS
金额：
$ 10.08万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-09-30 至 2002-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6027039
关键词：
aminoacid transport antisense nucleic acid cerebellar Purkinje cell cerebellar ataxia /dyskinesia chloride channels disease /disorder model gene induction /repression genetically modified animals glutamate transporter glutamates immunocytochemistry laboratory mouse laboratory rat neurogenetics neuronal transport oligonucleotides tissue /cell culture

项目摘要

Glutamate transport is essential for the synaptic inactivation of the neurotransmitter glutamate. A family of glutamate transporters has been identified in both astroglia and neurons. These transporters play important roles in the pathophysiology of neurologic disorders, notably ALS and Alzheimers disease. EAAT4 is a glutamate transporter selectively localized to cerebellar Purkinje cells and has the properties of a ligand-gated chloride ion channel. Its role in cerebellar physiology and in the possible pathogenesis of ataxic disorders is unknown. In this proposal, experiments are presented to study EAAT4 in Purkinje cell biology with larger implications for disorders to which EAAT4 dysfunction may contribute. First, a technique for the establishment of organotypic cerebellar cultures will be developed. These cultures will maintain their in vivo architecture and express the glutamate transporters, including EAAT4, that are present in the cerebellum. This system will allow the ability to manipulate conditions and assess factors that affect this glutamate transporter. Second, the phenotype that results from molecular knockdown of EAAT4 will be studied along with the neuroanatomical and neurophysiological changes in the cerebellum that accompany this reduction in protein expression. Finally, I will study whether EAAT4 dysfunction contributes to Purkinje cell death in the SCA1 transgenic mouse; a model for the human neurodegenerative disorder spinocerebellar ataxia.

谷氨酸转运对于神经递质谷氨酸的突触失活至关重要。在星形胶质细胞和神经元中都发现了谷氨酸转运蛋白家族。这些转运蛋白在神经系统疾病（尤其是 ALS 和阿尔茨海默病）的病理生理学中发挥着重要作用。 EAAT4 是一种选择性定位于小脑浦肯野细胞的谷氨酸转运蛋白，具有配体门控氯离子通道的特性。它在小脑生理学和共济失调疾病的可能发病机制中的作用尚不清楚。在本提案中，提出了在浦肯野细胞生物学中研究 EAAT4 的实验，这对 EAAT4 功能障碍可能导致的疾病具有更大的影响。首先，将开发建立器官型小脑培养物的技术。这些培养物将保持其体内结构并表达小脑中存在的谷氨酸转运蛋白，包括 EAAT4。该系统将能够操纵条件并评估影响谷氨酸转运蛋白的因素。其次，将研究 EAAT4 分子敲低所产生的表型，以及伴随蛋白质表达减少的小脑神经解剖学和神经生理学变化。最后，我将研究EAAT4功能障碍是否会导致SCA1转基因小鼠的浦肯野细胞死亡；人类神经退行性疾病脊髓小脑共济失调的模型。