A genetic indicator provides long term mapping of neuronal and calcium activity

遗传指标提供神经元和钙活动的长期图谱

• 批准号: 8078914
• 负责人: PAUL BREHM
• 金额: $ 30.49万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 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 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8078914
• 关键词: Adopted; Bioluminescence; Calcium; Calcium Binding; Calcium Signaling; Cells; Cloning; Code; Fluorescence; Genes; Genetic; Health; Histology; Image; Light; Maps; Measurement; Muscle Cells; Nerve; Neuraxis; Neurons; Property; Proteins; Reading; Recording of previous events; Resources; Secretory Cell; Signal Pathway; Tissues; cell type; in vivo; interest; luminescence; membrane activity

DESCRIPTION (provided by applicant): We are cloning a gene from an echinoderm that has the potential of representing the first long-term marker of calcium and neuronal activity. The gene encodes a protein that is responsible for the bioluminescence in certain species of brittlestars and has remarkable fluorescence properties. Only after calcium-triggered bioluminescence does this protein adopt a bright green fluorescence that persists indefinitely! The protein is present in nerves and it marks the active neurons in vivo for days after stimulated to produce luminescence and indefinitely if the tissue is fixed for histology. Like GFP, the fluorescence of this protein is intense and slow to bleach so it does not require low light level measurements. It has spectral properties very similar to GFP so that all of the resources developed for this standard can be used for ophiopsilin. Once cloned, we should be able to target the protein in any cell type of interest and examine the history of activity. Because virtually all cells use calcium signaling pathways it would serve as a universal indicator. Ideally suited for this type of approach are nerve, muscle and secretory cell types. It may be possible to use this indicator in the central nervous system to trace the circuitry at a single cell level. Thus, this calcium latch protein has the potential to revolutionize the manner in which we image the history of cellular activity in healthy and diseased tissue. Project Narrative We are exploiting the unique features of a naturally occurring calcium dependent fluorescence indicator in the neurons of echinoderms. The protein responsible for bioluminescence latches into a persistent GFP like fluorescence after binding calcium. As such it represents the first long-term marker of calcium and membrane activity. Because it is genetically coded it can be targeted for expression to any cell type and the fluorescence even persists through histology. As such it could be serve as a read out history of membrane activity and calcium signaling in healthy or diseased tissue. PUBLIC HEALTH RELEVANCE: We are exploiting the unique features of a naturally occurring calcium dependent fluorescence indicator in the neurons of echinoderms. The protein responsible for bioluminescence latches into a persistent GFP like fluorescence after binding calcium. As such it represents the first long-term marker of calcium and membrane activity. Because it is genetically coded it can be targeted for expression to any cell type and the fluorescence even persists through histology. As such it could be serve as a read out history of membrane activity and calcium signaling in healthy or diseased tissue.

描述(申请人提供)：我们正在从棘皮动物中克隆一种基因，该基因有可能代表第一个钙和神经元活动的长期标记物。该基因编码一种蛋白质，负责某些物种的生物发光，并具有显着的荧光特性。只有在钙引发的生物发光之后，这种蛋白质才会采用一种明亮的绿色荧光，这种荧光会无限期地持续下去！这种蛋白质存在于神经中，它标志着体内活跃的神经元在受到刺激后几天内产生发光，如果组织被组织学固定，它可以无限期地标记。像绿色荧光蛋白一样，这种蛋白质的荧光很强，漂白速度很慢，所以它不需要微光测量。它具有与GFP非常相似的光谱特性，因此为该标准开发的所有资源都可以用于蛇床子素。一旦克隆，我们应该能够针对任何感兴趣的细胞类型的蛋白质，并检查活动的历史。因为几乎所有的细胞都使用钙信号通路，所以它可以作为一个通用的指标。最适合这种方法的是神经、肌肉和分泌细胞类型。在中枢神经系统中使用这一指示器来追踪单个细胞水平的回路是可能的。因此，这种钙锁蛋白有可能彻底改变我们想象健康和疾病组织中细胞活动史的方式。项目简介我们正在利用棘皮动物神经元中一种自然产生的钙依赖荧光指示剂的独特功能。负责生物发光的蛋白质在与钙结合后锁定为持续的GFP样荧光。因此，它代表了第一个钙和膜活性的长期标记物。因为它是基因编码的，所以它可以针对任何类型的细胞表达，而且这种荧光甚至可以在组织学中持续存在。因此，它可以作为健康或患病组织中膜活性和钙信号的读出历史。与公共健康相关：我们正在利用棘皮动物神经元中自然产生的钙依赖荧光指示剂的独特功能。负责生物发光的蛋白质在与钙结合后锁定为持续的GFP样荧光。因此，它代表了第一个钙和膜活性的长期标记物。因为它是基因编码的，所以它可以针对任何类型的细胞表达，而且这种荧光甚至可以在组织学中持续存在。因此，它可以作为健康或患病组织中膜活性和钙信号的读出历史。