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

描述（由申请人提供）：我们正在克隆一种棘皮动物的基因，该基因有可能代表钙和神经元活动的第一个长期标记。该基因编码了一种蛋白质，这种蛋白质在某些种类的脆星中负责生物发光，并具有显著的荧光特性。只有在钙触发的生物发光之后，这种蛋白质才会发出明亮的绿色荧光，并无限期地持续下去！这种蛋白质存在于神经中，在刺激产生发光后的几天内，它会在体内标记活跃的神经元，如果组织在组织学上是固定的，它会无限期地标记活跃的神经元。与绿色荧光蛋白一样，这种蛋白质的荧光强度大，漂白速度慢，因此不需要在弱光下测量。它具有与绿色荧光蛋白非常相似的光谱特性，因此为本标准开发的所有资源都可以用于ophiopsilin。一旦克隆出来，我们应该能够针对任何感兴趣的细胞类型中的蛋白质并检查其活动历史。因为几乎所有的细胞都使用钙信号通路，所以它可以作为一种通用的指示物。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种指示器可以在单个细胞水平上追踪回路。因此，这种钙锁存蛋白有可能彻底改变我们在健康和病变组织中成像细胞活动历史的方式。我们正在利用棘皮动物神经元中天然存在的钙依赖荧光指示器的独特特征。负责生物发光的蛋白质在与钙结合后锁定为持久的GFP样荧光。因此，它代表了钙和膜活性的第一个长期标志。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，荧光甚至可以通过组织学持续存在。因此，它可以作为健康或病变组织中膜活性和钙信号传导的历史。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特征。负责生物发光的蛋白质在与钙结合后锁定为持久的GFP样荧光。因此，它代表了钙和膜活性的第一个长期标志。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，荧光甚至可以通过组织学持续存在。因此，它可以作为健康或病变组织中膜活性和钙信号传导的历史。

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.