CAGED PROBES FOR CELLULAR PHYSIOLOGY

描述（由申请人提供）：“笼状”分子是生物活性分子的光敏但暂时惰性的前体。光的吸收将前体转化为具有完全生物活性的分子。由于光可以很容易地聚焦和转向，而且光化学变化很快，笼状分子代表了一种多功能的工具，可以通过高时空控制来操纵生物学。该项目的长期目标是开发一种广谱的笼状分子，使细胞生理学家能够利用光来探测和控制活细胞和组织中的动态信号传导过程。该提案有四个重点：1）开发新的“笼”，其具有a）在单光子和双光子应用中的强光吸收，B）在光解时产物分子的高产率，c）产物光释放的快速动力学，和d）高的光解前稳定性。 2)开发新的神经信号的笼状探针，包括神经递质、脂质介质和递质转运抑制剂。3)开发两种新的细胞内钙信号特异性探针：最佳笼状钙和笼状兰尼碱受体激动剂。4)在正在进行的和新兴的独立和合作研究中应用新开发的工具，探索细胞神经生理学的四个主题：树突信号处理，突触可塑性，神经胶质细胞对突触事件的塑造，以及钙对兴奋性的调节。

DESCRIPTION (provided by applicant): A "caged" molecule is a photosensitive, but temporarily inert, precursor of a biologically active molecule. Absorption of light transforms the precursor into a molecule with full bioactivity. Because light can be easily focused and steered, and photochemical changes are fast, caged molecules represent a versatile tool for manipulating biology with light with high temporal and spatial control. The long-term objective of this project is to develop a broad spectrum of caged molecules that will allow cellular physiologists to use light to probe and control dynamic signaling processes in living cells and tissues. This proposal has four foci: 1) Develop new "cages" that have a) strong light absorption in single- and two-photon applications, b) high yield of product molecules upon photolysis, c) fast kinetics of product photorelease, and d) high pre-photolysis stability. 2) Develop new caged probes of neural signaling including neurotransmitters, lipid mediators, and inhibitors of transmitter transport. 3) Develop two new specific probes of intracellular calcium signaling: an optimal caged calcium and a caged agonist for the ryanodine receptor. 4) Apply the newly developed tools in ongoing and emerging independent and collaborative research to explore four topics in cellular neurophysiology: dendritic signal processing, synaptic plasticity, shaping of the synaptic event by glia, and calcium regulation of excitability.