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次外科手术和有害手术。虽然异丙酚的临床应用已经扩大，但在所有层面上对其许多作用的机械性理解都滞后了。在这项建议中，我们将部署我们的光活性烷基酚类似物异丙酚，azi-异丙酚-m(Azipm)和2-propynloxymethoxy-5-trifluoromethyldiazirinylphenol(azipm-Click)，在分层的一系列研究中揭示：使用最先进的质谱学和浓缩方法的光活性烷基酚结合蛋白质组(目标1)，光活性烷基酚在脑片离散核中的电生理特性(目标2)，以及个体细胞和网络在体内对烷基酚催眠作用的贡献。这些后一项研究将使用光纤激光刺激活体小鼠在全身注射光活性烷基酚后大脑的离散区域(目标3)。这三个目标通过他们对分子底物的共同预测而结合在一起，这些分子底物来自非常不同的、互补的和不偏不倚的方法。这些研究将确定烷基酚作用的新的和看似合理的分子底物，这将成为随后涉及构建转基因动物的提案中验证的基础。这一合作项目非常适合将凯尔茨实验室的神经生物学专业知识与贝克实验室的电生理学技能以及埃肯霍夫和戴利实验室的生物化学能力相结合。

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.