Expression of G-protein signaling components in adult mammalian neurons by microinjection.
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.
Ikeda
Stephen R
SR
Laboratory of Molecular Physiology, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD, USA.
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D018014:Gene Transfer Techniques; D008845:Microinjections / Q000379:methods*; D009474:Neurons / Q000378:metabolism*; D051381:Rats; D043562:Receptors, G-Protein-Coupled / Q000235:genetics* / Q000378:metabolism; D017783:Superior Cervical Ganglion / Q000378:metabolism
D018076:DNA, Complementary; D043562:Receptors, G-Protein-Coupled