PINP: a new method of tagging neuronal populations for identification during in vivo electrophysiological recording.
PINP: a new method of tagging neuronal populations for identification during in vivo electrophysiological recording.
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DOI：
10.1371/journal.pone.0006099
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发表时间：
2009-07-07
期刊：
影响因子：
3.7
通讯作者：
Zador AM
中科院分区：
文献类型：
--
作者：
Lima SQ;Hromádka T;Znamenskiy P;Zador AM
神经回路是由许多不同的神经元亚群组成的。然而，这是很难评估的功能作用，这些亚群使用传统的细胞外记录技术，因为这些技术不容易区分尖峰从不同的神经元群体。为了克服这一局限性，我们开发了PINP（光刺激辅助识别神经元群体），一种标记神经元群体的方法，用于在体内电生理记录过程中进行识别。该方法是基于表达光激活通道channelrhodopsin-2（ChR 2）的限制神经元亚群。ChR 2标记的神经元可以在体内电生理检测，因为用短暂的蓝光照射这些神经元会触发短潜伏期的可靠动作电位。我们通过使用两种不同的策略在不同的皮质神经元群体中表达ChR 2来证明这种技术的可行性。首先，我们标记了一个亚群的皮质神经元，主要是快速扣球interneurons，通过使用腺相关病毒（AAV）提供ChR 2的转基因小鼠系中的Cre重组酶的表达驱动的小清蛋白启动子。其次，我们标记的兴奋性神经元在大鼠听觉皮层与ChR 2的基础上的投影目标，通过使用单纯疱疹病毒1（HSV 1），这是有效地采取了轴突和运输逆行亚群，我们发现，后者人口响应声刺激不同的未标记的神经元。标记神经元是ChR 2的一种新应用，在这种情况下，用于监测活动，而不是操纵it. PINP可以很容易地扩展到其他群体的遗传可识别的神经元，并将提供一个有用的方法来探测不同的神经元群体在体内的功能作用。
Neural circuits are exquisitely organized, consisting of many different neuronal subpopulations. However, it is difficult to assess the functional roles of these subpopulations using conventional extracellular recording techniques because these techniques do not easily distinguish spikes from different neuronal populations. To overcome this limitation, we have developed PINP (Photostimulation-assisted Identification of Neuronal Populations), a method of tagging neuronal populations for identification during in vivo electrophysiological recording. The method is based on expressing the light-activated channel channelrhodopsin-2 (ChR2) to restricted neuronal subpopulations. ChR2-tagged neurons can be detected electrophysiologically in vivo since illumination of these neurons with a brief flash of blue light triggers a short latency reliable action potential. We demonstrate the feasibility of this technique by expressing ChR2 in distinct populations of cortical neurons using two different strategies. First, we labeled a subpopulation of cortical neurons—mainly fast-spiking interneurons—by using adeno-associated virus (AAV) to deliver ChR2 in a transgenic mouse line in which the expression of Cre recombinase was driven by the parvalbumin promoter. Second, we labeled subpopulations of excitatory neurons in the rat auditory cortex with ChR2 based on projection target by using herpes simplex virus 1 (HSV1), which is efficiently taken up by axons and transported retrogradely; we find that this latter population responds to acoustic stimulation differently from unlabeled neurons. Tagging neurons is a novel application of ChR2, used in this case to monitor activity instead of manipulating it. PINP can be readily extended to other populations of genetically identifiable neurons, and will provide a useful method for probing the functional role of different neuronal populations in vivo.
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影响因子：
16.2
作者：
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通讯作者：
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