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Recombinant Adeno-associated virus (rAAV)-mediated transduction and optogenetic manipulation of cortical neurons in vitro

机译:重组腺相关病毒(rAAV)介导的皮层神经元体外转导和光遗传学操作

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Genetically encoded light-sensitive proteins can be used to manipulate and observe cellular functions. According to different modes of action, these proteins are divided into actuators like the blue-light gated cation channel Channelrhodopsin-2 (ChR2) and detectors like the calcium sensor GCaMP. In order to optogenetically control and study the activity of rat primary cortical neurons, we established a transduction procedure using recombinant Adeno-associated viruses (rAAVs) as gene-ferries. Thereby, we achieved high transduction rates of these neurons with ChR2. In ChR2 expressing neurons, action potentials could be repeatedly and precisely elicited with laser pulses and measured via patch clamp recording.
机译:遗传编码的光敏蛋白可用于操纵和观察细胞功能。根据不同的作用方式,这些蛋白质分为致动器(例如,蓝光门控阳离子通道Channelrhodopsin-2(ChR2))和检测器(例如,钙传感器GCaMP)。为了光遗传学控制和研究大鼠原代皮层神经元的活性,我们建立了使用重组腺相关病毒(rAAVs)作为基因渡越的转导程序。因此,我们实现了这些神经元与ChR2的高转导率。在表达ChR2的神经元中,可以用激光脉冲反复精确地触发动作电位,并通过膜片钳记录进行测量。

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