Laboratory Course in the Molecular Biology of Excitable Membranes

可兴奋膜分子生物学实验课程

• 批准号: 9352275
• 负责人: Henry Lester
• 金额: $ 5.05万
• 依托单位: California Institute of Technology
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-07-15 至 1995-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9352275&HistoricalAwards=false
• 关键词: Laboratory; Course; Molecular; Biology; Excitable

9352275 Lester A new laboratory course in cellular and molecular neuroscience has been established that is specifically tailored to undergraduate instruction, and uses the latest techniques and concepts in the molecular biology of excitable cells. The course emphasizes (1) the molecular biological and fine- structure function of the "excitability proteins" of neuronal and other membranes, (2) the nature of the intracellular signaling pathways that are activated by cell-surface receptors, and (3) the nature of the intracellular signaling pathways that are activated by cell-surface receptors. Most experiments take place in 2 sessions. In the first session, students begin with a cDNA clone encoding an ion channel or receptor. The students use PCR to generate an optimal DNA, suitable for in vitro cRNA synthesis, and carry out the cRNA synthesis reaction. The students inject the cRNA into Xenopus oocytes. In the second session, the students perform electrophysiological and pharmacological measurements on the injected oocytes. The initial sessions concentrate on relatively simple manipulations such as intracellular recording; then advance to 2-electrode voltage clamp and, finally, to patch-clamp experiments. Provisions are made for maximal utilization of the requested equipment in undergraduate research projects during the terms when the course is not offered formally. ***

9352275 Lester一门新的细胞和分子神经科学实验课程已经建立，这是专门为本科教学量身定制的，并使用了可兴奋细胞分子生物学的最新技术和概念。本课程强调(1)神经元和其他细胞膜上“兴奋性蛋白”的分子生物学和精细结构功能，(2)细胞表面受体激活的细胞内信号通路的性质，以及(3)细胞表面受体激活的细胞内信号通路的性质。大多数实验在2个阶段进行。在第一节课中，学生从一个编码离子通道或受体的cDNA克隆开始。学生利用PCR生成适合体外合成cRNA的最佳DNA，并进行cRNA合成反应。学生们将cRNA注射到爪蟾卵母细胞中。在第二阶段，学生们对注射的卵母细胞进行电生理和药理学测量。初始阶段集中于相对简单的操作，如细胞内记录；然后进行双电极电压钳实验，最后进行膜片钳实验。在未正式开设本课程的学期中，为最大限度地利用所要求的设备进行本科生研究项目作出规定。＊＊＊