Expression of genetically encoded photosensors in retinal bipolar neurons

基因编码光电传感器在视网膜双极神经元中的表达

• 批准号: 7680017
• 负责人: GARY G MATTHEWS
• 金额: $ 19.5万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7680017
• 关键词: Action Potentials; Animals; Automobile Driving; Binding; Biological Assay; Biological Models; Bipolar Neuron; Blindness; Cations; Cells; Chimeric Proteins; Chloride Ion; Chlorides; DNA; Development; Disease; Drosophila genus; Engineering; Equipment; Exons; Fishes; Fluorescence; Fluorescence Microscopy; Genes; Genetic; Genetic Engineering; Goals; Halorhodopsins; Human; Ion Channel; Ions; Laboratories; Lead; Left; Light; Lighting; Metabotropic Glutamate Receptors; Methods; Modeling; Molecular; Monitor; Mus; Neurons; Nucleic Acid Regulatory Sequences; Optics; Oranges; Pan Genus; Papain; Pathway interactions; Photoreceptors; Preparation; Proteins; Regulatory Element; Reporter; Research; Rest; Retina; Retinal; Retinal Cone; Retinal Diseases; Signal Transduction; Sorbus; Synapses; Testing; Transgenes; Transgenic Animals; Transgenic Organisms; Translating; Universities; Variant; Vision; Visual; Visual system structure; Whole-Cell Recordings; Zebrafish; base; blind; functional restoration; light gated; mutant; novel strategies; patch clamp; photoreceptor degeneration; prevent; promoter; public health relevance; receptor; recombinase; research study; response; restoration; retinal bipolar neuron; retinal progenitor cell; selective expression; success; tool; transcription factor; transgene expression; visual information; visual process; visual processing

DESCRIPTION (provided by applicant): The photoreceptor cells of the retina carry out the essential task of translating light into electrical signals that can be passed on and analyzed by the rest of the visual system. Diseases that cause the photoreceptor cells to degenerate lead to irreversible blindness, because the remaining neurons of the retina are not sensitive to light and therefore cannot substitute for the loss of the photoreceptor cells. The goal of the proposed research is to induce sensitivity to light in the surviving retinal bipolar neurons, which would normally receive synaptic inputs from photoreceptors, as a strategy to restore useful vision after photoreceptor degeneration. The approach is to use genetic engineering to induce bipolar neurons to express two different light-activated ion channels, channelrhodopsin-2 and halorhodopsin, that will mimic the naturally occurring light response of ON and OFF bipolar cells, respectively. To drive expression in bipolar neurons, the project will make use of transgenic animals that incorporate DNA encoding channelrhodopsin-2 or halorhodopsin under the control of gene regulatory elements that will confer expression selectively in each of the two subtypes of bipolar neurons. The results will establish the feasibility of using these genetically encoded photosensor proteins as a functional replacement for missing photoreceptors in the diseased retina. PUBLIC HEALTH RELEVANCE: Degeneration of the photoreceptor cells in the retina is a leading cause of blindness, and at present, there is no treatment. The proposed research explores a new approach for restoration of vision despite the loss of photoreceptor cells. The strategy uses molecular engineering to make the remaining neurons of the retina sensitive to light, thereby replacing the function of the missing photoreceptors.

描述（由申请人提供）：视网膜的感光细胞执行将光转化为电信号的基本任务，这些电信号可以被视觉系统的其余部分传递和分析。导致光感受器细胞退化的疾病会导致不可逆的失明，因为视网膜上剩余的神经元对光不敏感，因此不能替代光感受器细胞的损失。本研究的目标是诱导存活的视网膜双极神经元对光的敏感性，这些神经元通常会接受来自光感受器的突触输入，作为在光感受器变性后恢复有用视力的策略。该方法是利用基因工程诱导双极神经元表达两种不同的光激活离子通道，通道视紫红质-2和盐视紫红质，它们将分别模拟开和关双极细胞自然发生的光反应。为了驱动双极神经元的表达，该项目将利用转基因动物，在基因调控元件的控制下，将DNA编码通道视紫红质-2或盐视紫红质结合起来，在双极神经元的两种亚型中选择性地赋予表达。该结果将确定使用这些基因编码的光感蛋白作为病变视网膜中缺失的光感受器的功能性替代品的可行性。