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: 7681205
• 负责人: PAUL BREHM
• 金额: $ 30.8万
• 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 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7681205
• 关键词: Adopted; Bioluminescence; Calcium; Calcium Binding; Calcium Signaling; Cells; Cloning; Code; Fluorescence; Genes; Genetic; Histology; Image; Light; Maps; Measurement; Muscle; Nerve; Neuraxis; Neurons; Property; Proteins; Reading; Recording of previous events; Resources; Secretory Cell; Signal Pathway; Tissues; cell type; in vivo; interest; luminescence; membrane activity; public health relevance

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（由申请人提供）：我们正在从棘皮动物中克隆一个基因，该基因有可能代表钙和神经元活动的第一个长期标记。该基因编码的蛋白质负责某些海蛇尾物种的生物发光，并具有显着的荧光特性。只有在钙触发生物发光后，该蛋白质才会呈现无限期持续的亮绿色荧光！该蛋白质存在于神经中，在刺激产生发光后数天内，它会在体内标记活跃的神经元，如果组织被固定用于组织学，则它会无限期地标记。与 GFP 一样，这种蛋白质的荧光强烈且漂白缓慢，因此不需要低光水平测量。它具有与 GFP 非常相似的光谱特性，因此为此标准开发的所有资源均可用于 ophiopsilin。一旦克隆，我们应该能够靶向任何感兴趣的细胞类型中的蛋白质并检查活性历史。因为几乎所有细胞都使用钙信号通路，因此它可以作为通用指标。最适合这种方法的是神经、肌肉和分泌细胞类型。或许可以在中枢神经系统中使用该指示器来追踪单细胞水平的电路。因此，这种钙锁蛋白有可能彻底改变我们对健康和患病组织中细胞活动历史的成像方式。项目叙述我们正在利用棘皮动物神经元中天然存在的钙依赖性荧光指示剂的独特特征。负责生物发光的蛋白质在与钙结合后会产生持久的 GFP 样荧光。因此，它代表了钙和膜活性的第一个长期标志物。因为它是基因编码的，所以它可以靶向表达到任何细胞类型，并且荧光甚至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动和钙信号传导的读出历史。公共健康相关性：我们正在利用棘皮动物神经元中天然存在的钙依赖性荧光指示剂的独特特征。负责生物发光的蛋白质在与钙结合后会产生持久的 GFP 样荧光。因此，它代表了钙和膜活性的第一个长期标志物。因为它是基因编码的，所以它可以靶向表达到任何细胞类型，并且荧光甚至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动和钙信号传导的读出历史。