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首页> 外文期刊>The European Journal of Neuroscience >Sindbis viral-mediated expression of Ca2+-permeable AMPA receptors at hippocampal CA1 synapses and induction of NMDA receptor-independent long-term potentiation.
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Sindbis viral-mediated expression of Ca2+-permeable AMPA receptors at hippocampal CA1 synapses and induction of NMDA receptor-independent long-term potentiation.

机译：Sindbis病毒介导的Ca2 +渗透性AMPA受体在海马CA1突触中的表达以及与NMDA受体无关的长期增强作用的诱导。

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摘要

Gene manipulation in order to artificially express a particular gene in neurons in the central nervous system is a powerful tool for the analysis of brain function. Sindbis viral vectors have been developed to express high levels of foreign genes in postmitotic brain neurons with little transfection of glial cells. In this study, we expressed the gene encoding the unedited GluR2 (GluR-B) subunit of the AMPA-type glutamate receptor that forms inwardly rectifying and Ca2+-permeable channels, in rat CA1 hippocampal neurons in slice cultures using Sindbis viral vectors. The pyramidal cell layer of the CA1 region was injected with recombinant Sindbis viruses encoding both enhanced green fluorescent protein (GFP) and unedited GluR2. The GFP fluorescence from CA1 neurons could be detected as early as 6 h and reached a maximal level about 48 h postinfection. The inwardly rectifying and Ca2+-permeable AMPA receptors were expressed in most CA1 pyramidal cells expressing GFP. These AMPA receptors expressed by gene transfer were involved in fast excitatory neurotransmission elicited by electrical stimulation of the Schaffer collaterals in the stratum radiatum. Tetanic stimulation of Schaffer collaterals induced NMDA receptor-independent, long-term potentiation due to Ca2+ influx through the newly expressed AMPA receptors in the area densely stained with GFP. Thus, the combined use of Sindbis viral vectors with the GFP reporter allowed physiological examination of the roles of a specific gene product in synaptic function in well-characterized brain neurons.
机译：为了在中枢神经系统的神经元中人工表达特定基因的基因操纵是分析脑功能的强大工具。已开发出Sindbis病毒载体，以在胶质细胞很少转染的情况下在有丝分裂后的脑神经元中表达高水平的外源基因。在这项研究中，我们使用Sindbis病毒载体在切片培养的大鼠CA1海马神经元中表达了编码AMPA型谷氨酸受体未编辑的GluR2（GluR-B）亚基的基因，该亚单位形成向内整流和Ca2 +渗透性通道。 CA1区的锥体细胞层注射了重组Sindbis病毒，该病毒同时编码增强的绿色荧光蛋白（GFP）和未经编辑的GluR2。来自CA1神经元的GFP荧光最早可在6小时被检测到，并在感染后48小时达到最大水平。在大多数表达GFP的CA1锥体细胞中表达了向内整流和Ca2 +可渗透的AMPA受体。这些通过基因转移表达的AMPA受体参与了由电刺激放射状的Schaffer侧支引起的快速兴奋性神经传递。对Schaffer侧支的强直刺激可引起NMDA受体非依赖性的长期增强，这是由于Ca2 +通过在GFP密集染色的区域中新表达的AMPA受体流入所致。因此，将Sindbis病毒载体与GFP报告基因结合使用，可以对特定基因产物在特征明确的脑神经元的突触功能中的作用进行生理检查。

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