Optoanesthesia

描述（由申请人提供）：烷基酚麻醉剂异丙酚每年用于数百万例外科手术和有害手术。虽然异丙酚的临床应用已经扩大，但在各个层面上对其许多作用的机理理解却落后了。在本提案中，我们将利用异丙酚的光活性烷基酚类似物，aziPm和2-丙基氧氧甲氧基-5-三氟甲基二氮基苯酚（aziPm-click），在一系列分层研究中揭示：利用最先进的质谱和富集方法研究光活性烷基酚结合蛋白质组（目的1），研究脑切片中离散核中光活性烷基酚的电生理特性（目的2），以及单个细胞和网络对烷基酚催眠作用的体内贡献。这些后一项研究将在系统给予光活性烷基酚后，对活小鼠的大脑离散区域进行光纤激光刺激（目的3）。这三个目标通过它们对分子底物的共同预测结合在一起，这些预测来自非常不同的、互补的和无偏的方法。这些研究将导致鉴定新的和合理的烷基酚作用的分子底物，这将作为验证后续涉及转基因动物构建的建议的基础。这个合作项目非常适合将Kelz实验室的神经生物学专业知识与Beck实验室的电生理学掌握以及Eckenhoff和Dailey实验室的生物化学实力相结合。

DESCRIPTION (provided by applicant): The alkylphenol anesthetic propofol is used in millions of surgical and noxious procedures annually. While the clinical use of propofol has swelled, mechanistic understanding of its many actions, on all levels, has lagged behind. In this proposal, we will deploy our photoactive alkylphenol analogues of propofol, azi-propofol-m (aziPm) and 2-propynloxymethoxy-5-trifluoromethyldiazirinylphenol (aziPm-click), in a tiered series of studies to reveal: the photoactive alkylphenol binding proteome using state-of-the-art mass spectrometry and enrichment methods (aim 1), the electro- physiological character of photoactive alkylphenols in discrete nuclei in brain slices (aim 2), and the in-vivo contribution o individual cells and networks to the hypnotic effects of alkylphenols. These latter studies will us fiberoptic laser stimulation of discrete regions of brain in live mice after systemic administratio of photoactive alkylphenols (aim 3). The three aims are integrated by their common prediction of molecular substrates arising from very different, complimentary and unbiased approaches. These studies will result in the identification of novel and plausible molecular substrates for alkylphenol action, which will serve as the basis for validation in subsequent proposals involving the construction of genetically altered animals. This collaborative project is ideally suited to integrate the neurobiological expertise of the Kelz lab with the electrophysiologic mastery of the Beck lab, and the biological chemistry prowess of the Eckenhoff and Dailey labs.