Elementary Events of Intracellular Calcium Signaling

钙离子在几乎所有类型的细胞中都是一种信号机制，调节着许多不同的功能。 从单个钙离子通道的开口(基本事件)开始， 通过集群通道(基本事件，如钙离子喷发)的协同开放 传播钙波，通过钙离子扩散和钙离子诱导的钙释放来协调。 因此，基本事件和基本事件构成了复杂结构的触发因素和构建块 允许对细胞功能进行分级和选择性调节的时空钙信号。我们的总体目标 是为了阐明钙离子通道的功能特性、空间组织和相互作用 时空细胞信号的模式。我们主要研究三磷酸肌醇(IP3)中的钙离子事件。 信号转导途径和钙离子通过淀粉样蛋白低聚物孔的通量在糖尿病的病理生理机制中的作用 阿尔茨海默氏症。利用生物光子学技术的最新进展，包括完全内部 反射荧光和超分辨显微镜，我们现在可以在真正单一的- 完整细胞中的分子水平。我们的目标是：(I)进一步开发快速三维成像技术 通过完整细胞中单个IPS受体/通道(IP3R)的钙离子流量；监测内质网[Ca2+]；以及 转基因小鼠模型中天然IP3Rs亚型的单分子定位 完整细胞中IP3R的功能特性，如何协调通道的活动来产生和 模板化基本的钙离子喷发，并产生全球钙离子波，(Iii)用 亚微米精度，以解决有关这些组件的集群组织和锚定的假设 通道；它们通过钙离子扩散和变构机制的相互作用；以及‘Silenf’的假定功能 膨化位点之间的IP3Rs，(Iv)研究淀粉样寡聚体形成钙离子的分子机制 单通道钙离子成像与单分子光漂白相结合的透气性毛孔 荧光单体测定九孔化学计量比，并阐明细胞内 低聚物通过IP3Rs诱导钙离子释放。 相关性(请参阅说明)： 钙离子在体内几乎所有的细胞中都起着“生或死”的作用，调节的过程多种多样。 心跳和脑细胞之间的突触传递，并与阿尔茨海默氏症和其他 疾病。我们的目标是阐明钙信号产生的分级机制。 单分子水平，具有更好地了解它们的正常功能和如何 信号转导MAV的r-alrJum基因缺失会导致疾病。

Ca2+ ions serve as a signaling mechanism in almost all cell types to regulate numerous diverse functions. Ca2+ signals are ordered in a hierarchy, from openings of single Ca2+ channels ('fundamental' events), through the concerted openings of clustered channels ('elementary' events, such as Ca2+ puffs) to propagating Ca2+ waves, coordinated through Ca2+ diffusion and Ca2+-induced Ca2+ release. Fundamental and elementary events thus form the triggers and building blocks underlying the complex spatiotemporal Ca2+ signals that permit graded and selective regulation of cell functions. Our overall goals are to elucidate how the functional properties, spatial organization and interactions between Ca2-t- channels pattern spatiotemporal cellular signals. We focus on Ca2+ events underlying the inositol trisphosphate (IP3) signaling pathway, and Ca2+ flux through amyloid oligomer pores implicated in the pathophysiology of Alzheimer's disease. Capitalizing on recent advances in biophotonic technology, including total intemal reflection fluorescence and superresolution microscopy, we can now study these topics at the truly single- molecule level in intact cells. Our aims are to: (i) Further develop techniques for fast, 3-dimensional imaging Ca2+ flux through individual IPS receptor/channels (IP3Rs) in intact cells; for monitoring ER [Ca2+]; and for single-molecule localization of subtypes of native IP3Rs in transgenic mouse models, (ii) Elucidate the functional properties of IP3Rs in the intact cell, how the activity of channels is orchestrated to generate and temiinate elementary Ca2+ puffs, and to generate global Ca2+ waves, (iii) Resolve IP3R molecules with sub-micron precision to address hypotheses concerning the clustered organization and anchoring of these channels; their interactions via Ca2+ diffusion and allosteric mechanisms; and the putative function of 'silenf IP3Rs between puff sites, (iv) Investigate the molecular mechanisms by which amyloid oligomers form Ca2+- permeable pores by combining single-channel Ca2+ imaging with single-molecule photobleaching of fluorescent monomers to detennine pore stoichiometry; and elucidate the mechanisms by which intracellular oligomers induce Ca2+ liberation through IP3Rs. RELEVANCE (See instructions): Calcium sen/es a 'life or death' function in virtually all cells ofthe body, regulating processes as diverse as the heartbeat and synaptic transmission between brain cells, and is implicated in Alzheimer's and other diseases. Our goal is to elucidate the hierarchical mechanisms by which calcium signals are generated at the single-molecule level, with the dual aims of better understanding their normal functioning and how disruntions in r-alrJum signaling mav lead to disease.