Analysis of cerebellar Purkinje cells using EAAT4 glutamate transporter promoter reporter in mice generated via bacterial artificial chromosome-mediated transgenesis
Analysis of cerebellar Purkinje cells using EAAT4 glutamate transporter promoter reporter in mice generated via bacterial artificial chromosome-mediated transgenesis
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DOI：
10.1016/j.expneurol.2006.08.016
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发表时间：
2007-01-01
影响因子：
5.3
通讯作者：
Rothstein, Jeffrey D.
中科院分区：
文献类型：
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作者：
Gincel, Dan;Regan, Melissa R.;Rothstein, Jeffrey D.
EAAT 4谷氨酸转运蛋白有助于调节兴奋性神经传递，并防止小脑中谷氨酸介导的兴奋性毒性。免疫组织化学和原位杂交先前已经确定了表达这种蛋白质的小脑细胞群。然而，这些方法并不适合于评估转运蛋白及其基因的动态调节，尤其是在活组织中。为了更好地研究EAAT4的表达和调控，我们产生了细菌人工染色体（BAC）启动子eGFP报告基因转基因小鼠。对转基因小鼠的组织学分析显示，EAAT 4启动子主要在浦肯野细胞中有活性，但也可以在出生后早期在其他神经元中适度检测到。EAAT 4启动子活性在非神经元细胞中不存在。从BAC转基因小鼠制备的小脑器官型切片培养物为研究活组织中转运蛋白和浦肯野细胞的表达和调节提供了独特的试剂。启动子活性与蛋白质表达的相关性使得EAAT 4 BAC启动子报告基因成为研究EAAT 4表达调控的有价值的工具。(c)2006年爱思唯尔公司All rights reserved.
The EAAT4 glutamate transporter helps regulate excitatory neurotransmission and prevents glutamate-mediated excitotoxicity in the cerebellum. Immunohistochemistry and in situ hybridization have previously defined a cerebellar cell population expressing this protein. These methods, however, are not well suited for evaluating the dynamic regulation of the transporter and its gene-especially in living tissues. To better study EAAT4 expression and regulation, we generated bacterial artificial chromosome (BAC) promoter eGFP reporter transgenic mice. Histological analysis of the transgenic mice revealed that the EAAT4 promoter is active predominantly in Purkinje cells, but can also be modestly detected in other neurons early postnatally. EAAT4 promoter activity was not present in non-neuronal cells. Cerebellar organotypic slice cultures prepared from BAC transgenic mice provided a unique reagent to study transporter and Purkinje cell expression and regulation in living tissue. The correlation of promoter activity to protein expression makes the EAAT4 BAC promoter reporter a valuable tool to study regulation of EAAT4 expression. (c) 2006 Elsevier Inc. All rights reserved.