Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition.
Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition.
复制标题
DOI：
10.3389/fnins.2018.00643
复制
发表时间：
2018
影响因子：
4.3
通讯作者：
Gao S
中科院分区：
文献类型：
--
作者：
Beck S;Yu-Strzelczyk J;Pauls D;Constantin OM;Gee CE;Ehmann N;Kittel RJ;Nagel G;Gao S
随着光门控离子通道视紫红质-2（ChR 2）的引入，细胞或活生物体的光遗传操纵在神经科学中得到了广泛应用。ChR 2是一种非选择性阳离子通道，非常适合于在神经元中去极化和诱发动作电位。然而，其钙（Ca 2+）渗透性和单通道电导低，并且对于某些应用，可能期望细胞内Ca 2+的更持久的增加。此外，需要一种有效的光门控钾（K+）通道，其可以快速抑制靶向神经元中的尖峰。考虑到Ca 2+和K+在细胞生理学中的重要性，光激活的Ca 2+渗透和K+特异性通道将是光遗传学工具箱中受欢迎的补充。在这里，我们描述了新的光门控Ca 2+渗透和K+特异性通道的工程融合细菌光活化腺苷酸环化酶的环核苷酸门控通道具有高渗透性的Ca 2+或K+，分别。优化的融合构建体在非洲爪蟾卵母细胞和大鼠海马神经元中表现出很强的光门控电导。这些结构也可以用来控制果蝇幼虫的运动，在运动神经元中表达时。照明导致身体收缩时，运动神经元表达的光敏感的Ca 2 +-渗透通道，和身体伸展时，表达的光敏感的K+通道，有效和可逆的麻痹幼虫。这些结构的进一步优化将需要在成年苍蝇的应用，因为这两个结构导致羽化失败时，在运动神经元中表达。
Optogenetic manipulation of cells or living organisms became widely used in neuroscience following the introduction of the light-gated ion channel channelrhodopsin-2 (ChR2). ChR2 is a non-selective cation channel, ideally suited to depolarize and evoke action potentials in neurons. However, its calcium (Ca2+) permeability and single channel conductance are low and for some applications longer-lasting increases in intracellular Ca2+ might be desirable. Moreover, there is need for an efficient light-gated potassium (K+) channel that can rapidly inhibit spiking in targeted neurons. Considering the importance of Ca2+ and K+ in cell physiology, light-activated Ca2+-permeant and K+-specific channels would be welcome additions to the optogenetic toolbox. Here we describe the engineering of novel light-gated Ca2+-permeant and K+-specific channels by fusing a bacterial photoactivated adenylyl cyclase to cyclic nucleotide-gated channels with high permeability for Ca2+ or for K+, respectively. Optimized fusion constructs showed strong light-gated conductance in Xenopus laevis oocytes and in rat hippocampal neurons. These constructs could also be used to control the motility of Drosophila melanogaster larvae, when expressed in motoneurons. Illumination led to body contraction when motoneurons expressed the light-sensitive Ca2+-permeant channel, and to body extension when expressing the light-sensitive K+ channel, both effectively and reversibly paralyzing the larvae. Further optimization of these constructs will be required for application in adult flies since both constructs led to eclosion failure when expressed in motoneurons.
登录
查看更多内容
影响因子：
25
作者：
Mahn M;Prigge M;Ron S;Levy R;Yizhar O
通讯作者：
Yizhar O
影响因子：
25
作者：
Lin, John Y.;Knutsen, Per Magne;Muller, Arnaud;Kleinfeld, David;Tsien, Roger Y.
通讯作者：
Tsien, Roger Y.
影响因子：
25
作者：
Kleinlogel, Sonja;Feldbauer, Katrin;Bamberg, Ernst
通讯作者：
Bamberg, Ernst
影响因子：
--
作者：
Gee, Christine E;Ohmert, Iris;Oertner, Thomas G
通讯作者：
Oertner, Thomas G
DOI：
10.1126/stke.2202004pl6
发表时间：
2004-02-12
期刊：
Science's STKE : signal transduction knowledge environment
影响因子：
--
作者：
McGuire, Sean E;Mao, Zhengmei;Davis, Ronald L
通讯作者：
Davis, Ronald L