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

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性的第一个长期标志物。该基因编码一种蛋白质，该蛋白质负责某些脆星属物种的生物发光，并具有显着的荧光特性。只有在钙触发生物发光后，这种蛋白质才采用无限期持续的明亮的绿色荧光！这种蛋白质存在于神经中，它在刺激后数天内标记体内的活性神经元，以产生发光，如果组织被固定用于组织学，则无限期标记。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，因此为该标准开发的所有资源都可以用于蛇蛋白。一旦克隆，我们应该能够在任何感兴趣的细胞类型中靶向蛋白质，并检查活性的历史。因为几乎所有的细胞都使用钙信号通路，它将作为一个通用的指标。理想地适合于这种类型的方法是神经，肌肉和分泌细胞类型。也许可以在中枢神经系统中使用这种指示剂来跟踪单个细胞水平的电路。因此，这种钙闩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。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功能。负责生物发光的蛋白质在结合钙后锁存为持久的GFP样荧光。因此，它代表了钙和膜活性的第一个长期标志物。因为它是遗传编码的，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，它可以用作健康或患病组织中膜活性和钙信号传导的读出历史。公共卫生相关性：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功能。负责生物发光的蛋白质在结合钙后锁存为持久的GFP样荧光。因此，它代表了钙和膜活性的第一个长期标志物。因为它是遗传编码的，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，它可以用作健康或患病组织中膜活性和钙信号传导的读出历史。

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.