CELLULAR AND MOLECULAR TARGETS OF GENERAL ANESTHETICS

描述（摘自申请书）：本拟议项目的目的是促进对全身麻醉药在突触前和突触后部位作用的细胞和分子机制的理解，特别强调异氟烷和丙泊酚。该计划由四个项目组成，通过科学和行政核心联系在一起。项目一将建立一组丙泊酚类似物对非洲爪蟾蝌蚪翻正反射丧失的定量构效关系，并与其对重组γ-氨基丁酸A（GABAA）受体的调节作用进行比较。在项目2中，异氟烷和丙泊酚对突触前囊泡融合和内吞作用的影响将在培养的海马神经元中进行研究，使用光学和生物化学技术。在项目3中，将使用取代的半胱氨酸可及性方法探索异氟烷和丙泊酚与GABAA受体中特定氨基酸残基的相互作用。科学和行政核心将与科学咨询委员会和特邀演讲人一起组织和管理该小组每年两次的会议，合成用于项目1和3研究的新型丙泊酚类似物，并在每个项目进行生理测量时测量实验盐水中存在的异氟烷和丙泊酚浓度。拟议的计划将集中在中枢突触的全身麻醉剂的作用的分子位点。这些信息将有助于进一步表征全身麻醉药复杂的体内药理学基础的多个靶点，并可能导致更安全的药物的开发。

DESCRIPTION (taken from application): The objective of this proposed program project is to advance understanding of the cellular and molecular mechanisms of action of general anesthetics at both pre- and postsynaptic sites, with particular emphasis on isoflurane and propofol. The program consists of four projects linked together by a Scientific and Administrative Core. In Project 1, quantitative structure-activity relations will be established among a group of propofol analogs for loss of righting reflex in Xenopus laevis tadpoles, and compared with that for the modulation of recombinant gamma-aminobutyric acidA (GABAA) receptors. In Project 2, the effects of isoflurane and propofol on presynaptic vesicle fusion and endocytosis will be studied in cultured hippocampal neurons, using both optical and biochemical techniques. In Project 3, interactions of isoflurane and propofol with specific amino acid residues in the GABAA receptor will be probed using the substituted cysteine accessibility method. The Scientific and Administrative Core will serve to organize and administer twice-yearly meetings of the group, together with a Scientific Advisory Board and guest speakers, synthesize novel analogs of propofol for study in Projects 1 and 3), and measure the concentrations of isoflurane and propofol present in the experimental salines at the time of the physiological measurements made in each project. The proposed program will focus on the molecular sites of action of general anesthetics at central synapses. Such information will aid in further characterization of the multiple targets underlying the complex in vivo pharmacology of general anesthetics, and may lead to the development of safer agents.