Ikeda Stephen R
Laboratory of Molecular Physiology, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD, USA.
Methods Mol Biol. 2004;259:167-81. doi: 10.1385/1-59259-754-8:167.
Although methods for expressing foreign proteins in clonal cell lines are well established, mature neurons remain a difficult preparation for the introduction of foreign genes. Microinjection is a reliable method for producing robust targeted expression in neurons that has advantages over conventional transfection/infection methodologies. Here, I describe procedures for expressing signaling proteins in adult rat sympathetic neurons by direct microinjection of cRNA and cDNA into the cytoplasm and nucleus, respectively. The methods are applicable to a wide variety of peripheral and central neuron preparations, as well as clonal cell lines.
虽然在克隆细胞系中表达外源蛋白的方法已经很成熟,但成熟神经元仍然是引入外源基因的困难材料。显微注射是一种在神经元中产生强大靶向表达的可靠方法,它比传统的转染/感染方法具有优势。在这里,我描述了分别通过将cRNA和cDNA直接显微注射到成年大鼠交感神经元的细胞质和细胞核中来表达信号蛋白的程序。这些方法适用于多种外周和中枢神经元材料以及克隆细胞系。