Malyshev A Y, Roshchin M V, Smirnova G R, Dolgikh D A, Balaban P M, Ostrovsky M A
Institute of Higher Nervous Activity and Neurophysiology of the Russian Academy of Sciences, Russian Federation.
Institute of Higher Nervous Activity and Neurophysiology of the Russian Academy of Sciences, Russian Federation.
Neurosci Lett. 2017 Feb 15;640:76-80. doi: 10.1016/j.neulet.2017.01.026. Epub 2017 Jan 16.
Optogenetics is a powerful technique in neuroscience that provided a great success in studying the brain functions during the last decade. Progress of optogenetics crucially depends on development of new molecular tools. Light-activated cation-conducting channelrhodopsin2 was widely used for excitation of cells since the emergence of optogenetics. In 2015 a family of natural light activated chloride channels GtACR was identified which appeared to be a very promising tool for using in optogenetics experiments as a cell silencer. Here we examined properties of GtACR2 channel expressed in the rat layer 2/3 pyramidal neurons by means of in utero electroporation. We have found that despite strong inhibition the light stimulation of GtACR2-positive neurons can surprisingly lead to generation of action potentials, presumably initiated in the axonal terminals. Thus, when using the GtACR2 in optogenetics experiments, its ability to induce action potentials should be taken into account. Our results also open an interesting possibility of using the GtACR2 both as cell silencer and cell activator in the same experiment varying the pattern of light stimulation.
光遗传学是神经科学中的一项强大技术,在过去十年中,它在研究大脑功能方面取得了巨大成功。光遗传学的进展关键取决于新分子工具的开发。自光遗传学出现以来,光激活阳离子传导通道视紫红质2被广泛用于细胞的激发。2015年,一个天然光激活氯离子通道GtACR家族被鉴定出来,它似乎是光遗传学实验中作为细胞沉默剂使用的非常有前途的工具。在这里,我们通过子宫内电穿孔法研究了在大鼠第2/3层锥体神经元中表达的GtACR2通道的特性。我们发现,尽管有强烈的抑制作用,但对GtACR2阳性神经元的光刺激出人意料地会导致动作电位的产生,推测是在轴突终末起始的。因此,在光遗传学实验中使用GtACR2时,应考虑其诱导动作电位的能力。我们的结果还开启了在同一实验中通过改变光刺激模式将GtACR既用作细胞沉默剂又用作细胞激活剂的有趣可能性。