Light-enabled identification of the neural substrates for alkylphenol anesthesia

描述(由申请人提供)：这个项目有助于我们通过使用光活性类似物异丙酚来帮助我们理解全身麻醉剂异丙酚的药理学。异丙酚诱导催眠的机制在分子和系统神经科学水平上仍不清楚。此外，异丙酚和其他全身麻醉药越来越被认为是神经发育和神经退行性疾病风险的媒介，有可能导致不可逆的表型。异丙酚中含有二氮杂环丙酚，在长波紫外线(~370 nm)照射下可发生光解反应，生成活性卡宾中间体。涉及体内局部刺激异丙酚的新技术将阐明导致这种全身麻醉剂产生昏迷的大脑区域。对异丙酚进行的全面的体外蛋白质组学实验将揭示可能有助于烷基酚麻醉的分子底物以及与这些麻醉剂化合物相关的急性和长期副作用。此外，通过光标记和串联质谱学技术鉴定异丙酚的蛋白质结合部位将有助于深入了解结合的特异性，从而提高我们对这些疏水小分子的药物作用的理解。通过解决这些目标，所获得的知识将增加我们对全身麻醉药在中枢神经系统中的广泛影响的认识，并将为未来的研究提供领域，以将这些广泛使用的药物的有害潜力降至最低。

DESCRIPTION (provided by applicant): This project serves to contribute to our understanding of the pharmacology of the general anesthetic propofol through the use of a photoactive analogue, meta-azi-propofol. The mechanisms of propofol-induced hypnosis remain unclear on both molecular and systems neuroscience levels. Further, propofol and other general anesthetics are becoming increasingly recognized as mediators of neurodevelopmental and neurodegenerative risks with the potential for causing irreversible phenotypes. Meta-azi-propofol contains a diazirine moiety that undergoes photolysis when exposed to long wave ultraviolet light (~370 nm) to create a reactive carbene intermediate. Novel techniques involving localized stimulation of meta-azi-propofol in vivo will elucidate brain regions that contribute to the unconsciousness produced by this general anesthetic. Comprehensive in vitro proteomic experiments with meta-azi-propofol will uncover molecular substrates that potentially contribute to alkylphenol anesthesia and the acute and long-term side effects associated with these anesthetic compounds. Further, the identification of protein binding sites of meta-azi-propofol through photolabeling and tandem mass spectrometry techniques will provide insight into binding specificity in order to improve our understanding of drug action by these small hydrophobic molecules. By addressing these aims, the knowledge gained will add to our appreciation for the widespread impact of general anesthetics within the central nervous system and will generate areas for future research to minimize the harmful potential of these widely used drugs.