Laboratory Course in the Molecular Biology of Excitable Membranes

9352275莱斯特一门新的细胞和分子神经科学实验室课程已经建立，这门课程专门为本科教学量身定做，并使用了可兴奋细胞的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的“兴奋性蛋白”的分子生物学和精细结构功能，(2)由细胞表面受体激活的细胞内信号通路的性质，以及(3)由细胞表面受体激活的细胞内信号通路的性质。大多数实验分两次进行。在第一节课中，学生们从编码离子通道或受体的cdna克隆开始。学生用聚合酶链式反应产生一种适合于体外合成cRNA的最佳DNA，并进行cRNA合成反应。学生们将cna注射到非洲爪哇卵母细胞中。在第二节课中，学生们对注射的卵母细胞进行电生理和药理学测量。最初的过程集中在相对简单的操作，如细胞内记录；然后进入两电极电压钳，最后是膜片钳实验。在课程没有正式开设的学期中，为最大限度地利用本科生研究项目中所需的设备做了准备。***

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. ***