CAGED PROBES FOR CELLULAR PHYSIOLOGY

描述(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全部生物活性的分子。由于光可以很容易地聚焦和操纵，而且光化学变化很快，笼子分子代表了一种具有高度时间和空间控制的用光来操纵生物的通用工具。该项目的长期目标是开发一种广泛的笼式分子，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过程。这项建议有四个重点：1)开发新的“笼子”，具有a)在单光子和双光子应用中的强光吸收，b)光解产物分子的高产量，c)产物光释放的快速动力学，以及d)高的预光解稳定性。2)开发新的笼式神经信号探针，包括神经递质、脂质介体和递质转运抑制物。3)开发两种新的细胞内钙信号特异性探针：一种理想的笼状钙离子和一种笼状兰尼定受体激动剂。4)将新开发的工具应用于正在进行的和新兴的独立和协作研究中，探索细胞神经生理学中的四个主题：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兴奋性的钙调节。

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.