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In vivo imaging of the light response in mouse retinal ganglion cells based on a neuronal activity-dependent promoter

机译:基于神经元活性依赖性启动子的小鼠视网膜神经节细胞光反应的体内成像

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摘要

Diseases of the retinal ganglion cells (RGCs) are an important cause of blindness, yet the light response of individual RGCs is difficult to assess in vivo, particularly in mammals, due to a lack of effective methods. We report a simple in vivo platform 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 electrophysio-logically 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 simple in vivo assessment of RGC function, which should help basic research into the mechanisms of RGC death and the development of treatments for diseases involving the RGCs. (C) 2019 The Authors. Published by Elsevier Inc.
机译:视网膜神经节细胞(RGCs)的疾病是失明的重要原因,但由于缺乏有效方法,难以评估个体RGC的光响应,特别是在哺乳动物中。我们在体内平台上报告了一种简单的平台,用于基于荧光报道标记的增强突触突触活性响应元素(E-SARE)对小鼠RGCS的光响应进行成像,所述突出的神经元活性 - 依赖性基因转录。当封装成AAV载体的E-SARE驱动的D2venus被注入静脉内术后,使用共聚焦眼镜检查,在单细胞分辨率下可见表达D2venus的光响应视网膜神经元。免疫组织学评估将大多数这些细胞作为RGCs鉴定。在RGC损伤的鼠模型中,D2venus阳性细胞的数量与光诱导的响应幅度和视力相关,在视觉皮层上测量电神经测量,表明载体可以用作工具评估RGCS中的可视功能。本文所述的平台允许简单的RGC功能进行体内评估,这应该有助于基本研究RGC死亡的机制以及涉及RGCS的疾病的治疗方法。 (c)2019年作者。 elsevier公司发布

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