Abstract

To perform optogenetic prosthetics of the retinal ganglion cell receptive field, a bicistronic genetic construct carrying the genes encoding the excitatory (channelrhodopsin-2) and inhibitory (Guillardia theta anion channelrhodopsin GtACR2) rhodopsins was created. A characteristic feature of this construct was the combination of these two genes with a mutant IRES insertion between them, which ensures the exact ratio of expression levels of the first and second genes in each transfected cell. Illumination of the central part of the neuron with light with a wavelength of 470 nm induced the action potential generation in the cell. Stimulation of the peripheral neuronal region with light induced the inhibition of action potential generation. Thus, using optogenetics methods, we simulated the ON–OFF interaction in the retinal ganglion cell receptive field. Theoretically, this construct can be used for optogenetic prosthetics of degenerative retina in the case of its delivery to the ganglion cells with lentiviral vectors.

中文翻译：

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双顺反子构建体，用于变性视网膜神经节细胞感受野的光遗传学修复。

摘要

为了进行视网膜神经节细胞接受场的光遗传学修复，双顺反子遗传构建体携带编码兴奋性（channelrhodopsin-2）和抑制性（Guillardia）的基因θ阴离子视紫红质（GtACR2）视紫红质被创建。该构建体的特征是这两个基因之间具有突变的IRES插入，从而确保了每个转染细胞中第一个和第二个基因表达水平的确切比例。用波长为470 nm的光照射神经元的中央部分会诱导细胞中动作电位的产生。用光刺激周围神经元区域诱导了动作电位产生的抑制。因此，使用光遗传学方法，我们模拟了视网膜神经节细胞感受野中的开-关相互作用。从理论上讲，该构建体在通过慢病毒载体递送至神经节细胞的情况下，可用于变性视网膜的光遗传学修复。