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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Fast noninvasive activation and inhibition of neural and network activity by vertebrate rhodopsin and green algae channelrhodopsin.
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Fast noninvasive activation and inhibition of neural and network activity by vertebrate rhodopsin and green algae channelrhodopsin.

机译：脊椎动物视紫红质和绿藻通道视紫红质快速无创激活并抑制神经和网络活动。

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摘要

Techniques for fast noninvasive control of neuronal excitability will be of major importance for analyzing and understanding neuronal networks and animal behavior. To develop these tools we demonstrated that two light-activated signaling proteins, vertebrate rat rhodopsin 4 (RO4) and the green algae channelrhodospin 2 (ChR2), could be used to control neuronal excitability and modulate synaptic transmission. Vertebrate rhodopsin couples to the Gi/o, pertussis toxin-sensitive pathway to allow modulation of G protein-gated inward rectifying potassium channels and voltage-gated Ca2+ channels. Light-mediated activation of RO4 in cultured hippocampal neurons reduces neuronal firing within ms by hyperpolarization of the somato-dendritic membrane and when activated at presynaptic sites modulates synaptic transmission and paired-pulse facilitation. In contrast, somato-dendritic activation of ChR2 depolarizes neurons sufficiently to induce immediate action potentials, which precisely follow the ChR2 activation up to light stimulation frequencies of 20 Hz. To demonstrate that these constructs are useful for regulating network behavior in intact organisms, embryonic chick spinal cords were electroporated with either construct, allowing the frequency of episodes of spontaneous bursting activity, known to be important for motor circuit formation, to be precisely controlled. Thus light-activated vertebrate RO4 and green algae ChR2 allow the antagonistic control of neuronal function within ms to s in a precise, reversible, and noninvasive manner in cultured neurons and intact vertebrate spinal cords.
机译：快速无创控制神经元兴奋性的技术对于分析和理解神经元网络和动物行为至关重要。为了开发这些工具，我们证明了两种光激活信号蛋白，脊椎动物大鼠视紫红质4（RO4）和绿藻Channelrhodospin 2（ChR2），可用于控制神经元兴奋性和调节突触传递。脊椎动物视紫红质耦合到Gi / o，百日咳毒素敏感的途径，以调节G蛋白门控的内向整流钾通道和电压门控的Ca2 +通道。在培养的海马神经元中，光介导的RO4激活可通过体树突状细胞膜的超极化减少ms内的神经元放电，当在突触前位点被激活时可调节突触传递和成对脉冲促进。相比之下，ChR2的体树突状激活足以使神经元去极化，以诱导立即的动作电位，该动作电位恰好跟随ChR2激活直至20 Hz的光刺激频率。为了证明这些构建体可用于调节完整生物体中的网络行为，可对任一种构建体的胚胎小鸡脊髓进行电穿孔，从而精确控制自发爆发活动的发作频率，而自发爆发活动的频率对于形成运动回路很重要。因此，光激活的脊椎动物RO4和绿藻ChR2能够以精确，可逆且无创的方式在培养的神经元和完整的脊椎动物脊髓中对ms内的神经元功能进行拮抗控制。

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