Central synapse function studies with 2-photon uncaging

描述(申请人提供)：神经递质与特定的 完整的膜受体导致细胞反应的多样性。这个 这些不同反应的空间范围可能会有很大的差异。抑制性 神经元躯体上的输入可以对电信号产生显著的影响 整个单元格的属性。相反，1P3介导的细胞内释放 钙的储存可以被限制在单个树突棘突上。因此，空间 定位是突触化学信号传递的一个特别重要的特征 它被认为是某些形式的输入专一性的原因 突触可塑性(即长时程增强或抑制)。更多 一般说来，空间局部化的信号是广泛的神经元的基础。 在发育和成年期中的作用，例如细胞迁移、轴突引导和 神经计算。这些过程中的许多过程在病理上受到干扰 各州。在对这些疾病状态进行充分描述之前， 应该对非疾病状态有更全面的了解。这个 拟议的研究将有助于更好地理解正常 这些过程的运作。 这笔赠款的长期目标是开发和应用新的 设计用于神经递质的高度局域化光释放的发色团 突触。膜片钳技术和膜片钳技术的发展 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控细胞内的 实时监控环境。笼状化合物(即生物上的光敏化合物 惰性信号分子)补充这些电学和光学技术 因为它们在时间和空间上提供了对细胞化学的控制 域名。在过去的十年里，一种新型的固体激光技术 已经变得容易获得(钛：蓝宝石)，这允许红外 紫外光吸收发色团的激发。去中心化是由同步产生的 能量相等的两个红光子对一个蓝光子的吸收。然而， 目前可用的笼状化合物对双光子光解不敏感。 这笔赠款的目的是开发和测试新的发色团， 对于双光子光解是有效的。使用这些化合物的去除将 为生物体内神经递质的释放提供三维空间控制 细胞。有四个关于神经元功能的一般主题 将研究：1.受体定位；2.受体密度；3.受体 4.突触可塑性。这个项目将进一步推动我们的 了解大脑和意志中信号的基本机制 为了解神经系统疾病的病因铺平了道路 通过改变中央突触传递来采取行动。

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.