Optogenetic Approaches to Restore Vision and to Analyse Neural Circuits in the Retina
恢复视力和分析视网膜神经回路的光遗传学方法
基本信息
- 批准号:394730111
- 负责人:
- 金额:--
- 依托单位:
- 依托单位国家:德国
- 项目类别:Research Grants
- 财政年份:2018
- 资助国家:德国
- 起止时间:2017-12-31 至 2020-12-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The introduction of microbial opsins is a promising approach to restore light sensitivity in retinal degenerative diseases. The goal of this project is to express a red-light sensitive channelrhodopsin-variant in ON-bipolar cells of the primate retina, ex vivo by using a viral vector. Here, we will focus on the ON-bipolar cells of the central retina (ON-midget BPCs). A key feature of these cells is that they receive synaptic input from single cone photoreceptors, and these signals are then transmitted to single ganglion cells (midget RGCs). This one-to-one connectivity is an important anatomical prerequisite for the foveal high-acuity vision in primates, including humans. By using optogenetic stimulation of ON-midget BPCs we will trigger light-response in connected ganglion cells which will be analysed with electrophysiology. In particular, we will determine their receptive field properties which will provide important information about the potential spatial resolution that can be achieved with this optogenetic approach. In a parallel project, we will develop a tandem-construct, composed of a chlorid-pump/sensor that will allow for simultaneous manipulation and observation of intracellular chloride concentrations. Cell-type specific expression of this construct will enable to control the intracellular chloride concentration of these cells in a quantitative manner (ratiometric sensor), in order to investigate the impact of transient inhibition on local neuro-circuits. Therefore, we will use a custom-made setup that allows to stimulate retinal cones with patterned light at visible wavelengths, combined with 2-photon induced optogenetic (holographic) stimulation and 2-photon imaging.
引入微生物视蛋白是恢复视网膜退行性疾病的光敏性的一种很有前途的方法。本项目的目标是利用病毒载体在灵长类动物视网膜的ON-bipolar细胞中体外表达一种红光敏感通道视紫红质变体。在这里,我们将重点关注中央视网膜的on -双极细胞(on -侏儒BPCs)。这些细胞的一个关键特征是它们接受来自单个锥状光感受器的突触输入,然后这些信号被传递到单个神经节细胞(小RGCs)。这种一对一的连接是包括人类在内的灵长类动物中央凹高灵敏度视觉的重要解剖学先决条件。通过对ON-midget BPCs进行光遗传刺激,我们将在连接的神经节细胞中触发光反应,这将用电生理学进行分析。特别是,我们将确定它们的感受野特性,这将提供关于这种光遗传学方法可以实现的潜在空间分辨率的重要信息。在一个平行项目中,我们将开发一个串联结构,由一个氯泵/传感器组成,可以同时操作和观察细胞内氯浓度。这种结构的细胞类型特异性表达将能够定量地控制这些细胞的细胞内氯化物浓度(比率传感器),以便研究短暂抑制对局部神经回路的影响。因此,我们将使用一个定制的设置,允许在可见光波段的图案光刺激视网膜锥细胞,结合双光子诱导光遗传(全息)刺激和双光子成像。
项目成果
期刊论文数量(0)
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Dr. Jens Dübel其他文献
Dr. Jens Dübel的其他文献
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