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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Recombinant Semliki Forest virus and Sindbis virus efficiently infect neurons in hippocampal slice cultures
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Recombinant Semliki Forest virus and Sindbis virus efficiently infect neurons in hippocampal slice cultures

机译：重组Semliki Forest病毒和Sindbis病毒有效感染海马切片培养物中的神经元

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摘要

Gene transfer into nervous tissue is a pow- erful tool for the analysis of gene function. By using a rat hippocampal slice culture preparation, we show here that Semliki Forest virus (SFV) and Sindbis virus (SIN) vectors are useful for the effective infection of neurons. The stratum pyramidale and/or the granular cell layer were injected with recombinant virus encoding #beta#-galactosidase (LacZ) or green fluorescent protein (GFP). By using low concentrations of injected SFV-LacZ or SIN-LacZ, we detected LacZ staining of pyramidal cells, interneurons, and granule cells. About 60 of the infected cells showed clear neuronal morphology; thus, relatively few glial cells expressed the transgene. Expression of GFP from SFV and SIN vectors gave similar results, with an even higher percentage (>90) of the GFP-positive cells identified as neurons. Infected pyramidal cells were readily recognized in living slices, displaying GFP fluorescence in dendrites of up to fourth order and in dendritic spines. They appeared morphologically normal and viable at 1-5 days postinfection. We conclude that both SFV and SIN vectors efficiently transfer genes into neurons in hippocampal slice cultures. In combination with the GFP reporter, SFV and SIN vectors will allow the physiological examination of identified neurons that have been modified by overexpression or sup- pression of a specific gene product.
机译：基因转移到神经组织中是分析基因功能的有效工具。通过使用大鼠海马切片培养物，我们在此处显示了Semliki森林病毒（SFV）和Sindbis病毒（SIN）载体可用于有效感染神经元。注射编码＃beta＃-半乳糖苷酶（LacZ）或绿色荧光蛋白（GFP）的重组病毒，对角锥体层和/或颗粒细胞层进行注射。通过使用低浓度的注射SFV-LacZ或SIN-LacZ，我们检测了锥体细胞，中间神经元和颗粒细胞的LacZ染色。大约60个被感染的细胞显示出清晰的神经元形态。因此，相对较少的神经胶质细胞表达转基因。来自SFV和SIN载体的GFP表达给出了相似的结果，被鉴定为神经元的GFP阳性细胞百分比更高（> 90）。感染的锥体细胞很容易在活切片中识别出来，在最多四阶的树突和树突棘中显示GFP荧光。它们在感染后1-5天在形态上看来是正常的并且是可行的。我们得出的结论是，SFV和SIN载体均有效地将基因转移到海马切片培养物中的神经元中。结合GFP报告基因，SFV和SIN载体可对已鉴定的神经元进行生理检查，这些神经元已被特定基因产物的过表达或表达抑制。

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