Fast noninvasive activation and inhibition of neural and network activity by vertebrate rhodopin and green algae channelrhodopsin
Fast noninvasive activation and inhibition of neural and network activity by vertebrate rhodopin and green algae channelrhodopsin
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DOI：
10.1073/pnas.0509030102
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发表时间：
2005-12-06
影响因子：
11.1
通讯作者：
Herlitze, S
中科院分区：
文献类型：
--
作者：
Li, X;Gutierrez, DV;Herlitze, S
神经元兴奋性的快速非侵入性控制技术对于分析和理解神经元网络和动物行为具有重要意义。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（1104）和绿色藻类通道rhodospin 2（ChR 2），可用于控制神经元的兴奋性和调节突触传递。脊椎动物视紫红质与Gi/o百日咳毒素敏感通路偶联，以调节G蛋白门控内向整流钾通道和电压门控Ca 2+通道。光介导的激活RO 4在培养的海马神经元减少神经元放电ms内的体细胞树突膜的超极化，并在突触前位点激活时调制突触传递和成对脉冲促进。相反，ChR 2的体树突激活使神经元充分去极化以诱导即时动作电位，其精确地跟随ChR 2激活直到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，用任一构建体电穿孔胚胎鸡脊髓，允许精确控制已知对运动回路形成重要的自发爆发活动的发作频率。因此，光激活的脊椎动物RO 4和绿色藻类ChR 2允许在培养的神经元和完整的脊椎动物脊髓中以精确的、可逆的和非侵入性的方式在ms至s内拮抗性地控制神经元功能。
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 (1104) 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.