Central synapse function studies with 2-photon uncaging

描述（由申请人提供）：神经递质与特异性 完整的膜受体导致多种细胞反应。的 这些不同响应的空间范围可能差别很大。抑制 对神经元胞体的输入可以对电生理产生显著的影响。 整个细胞的特性。相反，1 P3介导的细胞内 钙的储存可限于单个树突棘。因此，空间 定位是突触化学信号传导的一个特别重要的特征 它被认为是某些形式的输入特异性的原因 突触可塑性（即长时程增强或抑制）。更 一般来说，空间定位的信号传导是广泛的神经元信号传导的基础。 在发育和成年期的功能，例如细胞迁移，轴突导向和 神经计算这些过程中有许多在病理上受到干扰， states.在充分描述这些疾病状态之前， 应更全面地了解非疾病状态。的 建议的研究将有助于更好地了解正常的 这些过程的运作。 这项赠款的长期目标是开发和应用新的 为神经递质的高度局部化光释放而设计的发色团. 突触膜片钳技术等技术的发展， 激光扫描共聚焦显微镜彻底改变了我们对许多 细胞功能。这些技术使我们能够监测细胞内 真实的环境。笼状化合物（即光敏、生物 惰性信号分子）补充了这些电学和光学技术 因为它们在时间和空间上提供细胞化学的控制， 域.在过去的十年中，一种新型的固体激光技术 已经变得容易获得（钛：蓝宝石），这使得红外 紫外吸收发色团的激发。解除锁定是由同时 吸收两个能量与一个蓝色光子相等的红色光子。然而，在这方面， 目前可用的笼状化合物对2-光子光解不敏感。 这是这个赠款的目的是开发和测试新的发色团， 对双光子光解有效。使用这些化合物将 提供三维空间控制的神经递质释放到生活 细胞关于神经元的功能，有四个一般性的主题， 将研究：1.受体定位; 2.受体密度; 3.受体 贩运; 4.突触可塑性该项目将进一步促进我们的 了解大脑和意志内部信号的基本机制 为理解神经系统疾病的病因铺平了道路， 通过改变中枢突触传递而起作用。

DESCRIPTION (provided by applicant): Neurotransmitter binding to specific integral membrane receptors results in a diversity of cellular responses. The spatial extent of these different responses may vary tremendously. Inhibitory inputs on the somata of neurons can have dramatic effects on the electrical properties of the whole cell. Conversely, 1P3 mediated release of intracellular stores of calcium can be limited to a single dendritic spine. Thus, spatial localization is an especially crucial feature of synaptic chemical signaling and it is believed to be responsible for the input specificity of certain forms of synaptic plasticity (viz, long-term potentiation or depression). More generally, spatially localized signaling underlies a wide range of neuronal function in development and adulthood, e.g. cell migration, axon guidance and neural computation. Many of these processes are disturbed in pathological states. Before an adequate description of these disease states can be given, a more complete understanding of non-disease states should be accomplished. The proposed studies will contribute to a greater understanding of the normal functioning of these processes. The long-term goal of this grant is the development and application of new chromophores designed for highly localized photorelease of neurotransmitters at synapses. The development of technologies such as the patch clamp technique and laser-scanning confocal microscopy has revolutionized our understanding of many cellular functions. These techniques enable us to monitor the intracellular environment in real time. Caged compounds (i.e. photosensitive, biologically inert signaling molecules) complement these electrical and optical techniques as they provide control of cellular chemistry, in both temporal and spatial domains. During the past ten years a new type of solid-state laser technology has become readily available (Ti:sapphire), which allows for infrared excitation of UV-absorbing chromophores. Uncaging is produced by simultaneous absorption of two red photons of equivalent energy to one blue photon. However, the currently available caged compounds are insensitive to 2-photon photolysis. It is the intent of this grant to develop and test new chromophores that will be effective for 2-photon photolysis. Uncaging using these compounds will provide 3-D spatial control of the release of neurotransmitters onto living cells. There are four general themes concerning the functioning of neurons that will be studied: 1. receptor localization; 2. receptor density; 3. receptor trafficking; and 4. synaptic plasticity. This project will further our understanding of the basic mechanisms of signaling within the brain and will pave the way toward understanding the etiology of neurological disorders that act by altering the central synaptic transmission.