In vivo imaging of the light response in mouse retinal ganglion cells based on a neuronal activity-dependent promoter
In vivo imaging of the light response in mouse retinal ganglion cells based on a neuronal activity-dependent promoter
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基于神经元活动依赖性启动子的小鼠视网膜神经节细胞光响应的体内成像
DOI：
10.1016/j.bbrc.2019.10.155
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发表时间：
2020
影响因子：
3.1
通讯作者：
Nakazawa Toru
中科院分区：
文献类型：
--
作者：
Fujita Kosuke;Nishiguchi Koji M.;Sato Kota;Nakagawa Yurika;Nakazawa Toru
视网膜神经节细胞（retinal ganglion cells，RGC）的疾病是致盲的重要原因，但由于缺乏有效的方法，难以在体内评估单个RGC的光反应，特别是在哺乳动物中。我们报告了一个简单的体内平台，用于成像小鼠RGC的光响应，该平台基于荧光标记的增强型突触活性响应元件（E-SARE），该元件介导神经元活性依赖性基因转录。当E-SARE驱动的d2 Venus包装成AAV载体，玻璃体内注射时，表达d2 Venus的光响应性视网膜神经元使用共聚焦检眼镜以单细胞分辨率可见。免疫组织学评估将这些细胞中的大多数鉴定为RGC。在RGC损伤的小鼠模型中，d2 Venus阳性细胞的数量与光诱导反应的幅度和视觉敏锐度相关，在视觉皮层电生理学测量，表明该载体可用作评估RGC视觉功能的工具。本文所述的平台允许RGC功能的简单体内评估，这将有助于对RGC死亡机制的基础研究和涉及RGC的疾病的治疗的开发。
Diseases of the retinal ganglion cells (RGCs) are an important cause of blindness, yet the light response of individual RGCs is difficult to assessin vivo,particularly in mammals, due to a lack of effective methods. We report a simplein vivoplatform for imaging the light response of mouse RGCs based on a fluorescent reporter-tagged enhanced synaptic activity-responsive element (E-SARE) that mediates neuronal activity–dependent gene transcription. When E-SARE-driven d2Venus, packaged into an AAV vector, was injected intravitreally, light-responsive retinal neurons expressing d2Venus were visible at single-cell resolution using confocal ophthalmoscopy. Immunohistological assessment identified the majority of these cells as RGCs. In a murine model of RGC injury, the number of d2Venus-positive cells was correlated with the amplitude of light-induced responses and with visual acuity, measured electrophysiologically at the visual cortex, indicating that the vector can be used as a tool to assess visual function in RGCs. The platform described herein allows a simplein vivoassessment of RGC function, which should help basic research into the mechanisms of RGC death and the development of treatments for diseases involving the RGCs.