Photochemical regulation of calcium in cell physiology

描述(由申请人提供)：提案的目标是开发用于控制细胞内信号通路的新的光学探针。在过去的15年里，我们制造了许多光学探针，我们称之为笼状化合物，在细胞生理学领域的所有此类探针中得到了广泛的应用。例如，我们开发的笼式钙探针已经被许多研究多种细胞类型的生理学的实验室用于数百个实验。最近，在这笔赠款的支持下开发的笼子里的神经递质产生了广泛的影响。任何技术要想真正具有变革性，不仅必须解决重大的未得到满足的需求，而且还必须真正起作用，必须是实用的。为了将我们的技术创新转化为现实，我们采取了多学科的方法， 将合成有机化学与光化学、激光光物理和细胞生理学相结合。我们工作的另一个重要特点是我们与著名生理学家建立了长期的合作关系。这些相互作用对有用和重要的笼状化合物的发展至关重要，因为它是定义探针的生物学问题。在这项建议中，我们试图解决用于光化学探测细胞内信号通路的光学化学方法领域的一个重要空白，即完成并行信号过程的同时、多模式光学控制的能力。大多数(但不是全部)新的探针将涉及细胞内钙的控制。钙变化的影响的时间和空间尺度是极其广泛的，所以解决所有这些水平的光学方法在许多类型的细胞生理学的研究中是有用的。例如，突触传递使用快速的局部钙变化，而钙调节的基因表达使用更持续的细胞钙变化。在心肌细胞中，钙的快速局部变化通过钙诱导的钙释放转化为全局钙释放事件，而钙诱导钙释放催化收缩。由于钙离子的重要性，微妙地调控钙离子本身或利用钙离子在一定程度上调控由钙离子控制的平行信号通路，往往是共生信号系统的结合点。这项提议的总体目标是开发新的光化学工具，允许对这些信号通路进行双向光学控制。 公共卫生相关性：光学方法是细胞生理学研究的主要技术。这项研究涉及到新的光学探针的开发，它将首次实现对两个细胞内信号系统的同时光控制。由于几乎所有的细胞信号都是双向的或共生的，这些新的方法将彻底改变我们控制功能的能力，从而揭示细胞生理学的许多新细节。

DESCRIPTION (provided by applicant): The goal of proposal is the development of new optical probes for controlling intracellular signaling pathways. Over the past 15 years we have made many optical probes which we call caged compounds that have been the widely of all such probes in the field of cell physiology. For example, the caged calcium probes we have developed have been used in hundreds of experiments by many laboratories studying the physiology of many cell types. More recently, the caged neurotransmitters developed under the aegis of this grant have had wide impact. For any technology to be truly transformative it must not only address a significant unmet need, but it must also really work, it must be practical. In order to translate our technological innovations into reality we take a multidisciplinary approach, combining synthetic organic chemistry and photochemistry, laser photophysics, and cellular physiology. An additional vital feature of our work is that we have forged long-term collaborative relationships with noted physiologists. These interactions have been vital the development of useful and important caged compounds as it is the biological problems that define the probes. In this proposal we seek to address an important gap in the area of optical chemical methods that are used for photochemical probing of intracellular signaling pathways, namely, the ability to accomplish simultaneous, multimodal optical control of parallel signaling processes. The majority, but not all, of the new probes will involve the control of intracellular calcium. The temporal and spatial scales of the effects of changes in calcium are extremely wide, so optical methods that address all these levels are useful in the study of many types of cellular physiology. For example, synaptic transmission uses fast, local changes in calcium, whereas calcium-regulated gene expression uses more sustained changes in cellular calcium. In cardiac myocytes, fast local changes in calcium are converted into global calcium release events, via calcium-induced calcium release, which catalyze contraction. Because calcium is so important, it often is the integration point of symbiotic signaling systems that subtlety regulates calcium itself or use calcium to modulate parallel signaling pathways that are controlled by calcium at some level. The overall goal of this proposal is to develop new photochemical tools that allow bidirectional optical control of these signaling pathways. PUBLIC HEALTH RELEVANCE: Optical methods are a primary technique for the study of cellular physiology. This research concerns the development of new optical probes that will enable simultaneous photocontrol of two intracellular signaling systems for the first time. Since almost all cell signaling is bidirectional or symbiotic, these new methods will revolutionize our ability to control function and therefore uncover many fresh details of cell physiology.