Kinetic profiles of photocurrents in cells expressing two types of channelrhodopsin genes.
Kinetic profiles of photocurrents in cells expressing two types of channelrhodopsin genes.
复制标题
表达两种视紫红质通道蛋白基因的细胞中光电流的动力学曲线。
DOI：
10.1016/j.bbrc.2018.01.149
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Tomita H.
中科院分区：
文献类型：
--
作者：
Watanabe Y;Sugano E;Tabata K;Ozaki T;Saito T;Tamai M;Tomita H.
视紫红质-2（ChR 2）是一种光激活的阳离子选择性离子通道，已被广泛用作光遗传学研究的工具。ChR 2对小于550 nm的波长特别敏感。扩大通道视紫红质对更宽波长范围的敏感性的方法之一是通过转导另外的基因在细胞中表达另一种通道视紫红质。在这里，我们报告的特性功能的细胞表达两种类型的通道视紫红质，每种具有不同的波长敏感性。在稳定表达ChR 2的HEK 293细胞中，在400-550 nm的刺激下引发光电流，并且通过额外转导修饰的团藻通道视紫红质-1（mVChR 1）基因来扩大波长敏感性范围，所述基因具有宽波长敏感性，范围从400至600 nm。然而，在550 nm处的光电流低于themVChR 1表达细胞，而且，打开和关闭常数延迟，失活速率降低。同时，外源基因的导入提高了番茄对弱光的反应。因此，额外基因的转导是改善通道视紫红质的光和波长灵敏度以及光电流动力学曲线的有用方法。
Channelrhodopsin-2 (ChR2), a light-activated cation-selective ion channel, has been widely used as a tool in optogenetic research. ChR2 is specifically sensitive to wavelengths less than 550 nm. One of the methods to expand the sensitivity of a channelrhodopsin to a wider range of wavelengths is to express another channelrhodopsin in the cells by the transduction of an additional gene. Here, we report the characteristic features of cells expressing two types of channelrhodopsins, each having different wavelength sensitivities. In HEK293 cells stably expressing ChR2, photocurrents were elicited at stimuli of 400–550 nm, and the wavelength sensitivity range was expanded by the additional transduction of the modified Volvox channelrhodopsin-1 (mVChR1) gene, which has broad wavelength sensitivities, ranging from 400 to 600 nm. However, the photocurrent at 550 nm was lower than that of themVChR1-expressing cell; moreover, the turning-on and turning-off constants were delayed, and the deactivation rates were decreased. Meanwhile, the response to lower light intensity was improved by the additional gene. Thus, the transduction of an additional gene is a useful method to improve the light and wavelength sensitivities, as well as photocurrent kinetic profiles, of channelrhodopsins.