In situ hybridization analysis of RGS mRNA regulation and behavioral phenotyping of RGS mutant mice.
To elucidate the functional role of regulators of G-protein signaling (RGS) in vivo, it will be critical to (i) determine how RGS activity is altered in response to a variety of manipulations and (ii) observe how the system is changed when RGS protein function is altered genetically. To facilitate studies of dynamic regulation of RGS protein activity, this article describes detailed methods for radioisotopic in situ hybridization for semiquantitative analyses of RGS mRNA abundances. Toward characterizing the functional differences in mice with genetically altered RGS activities, this article describes a subset of behavioral tests suitable for assaying sensitivities to drugs of abuse. These protocols should provide valuable guidance for investigators to establish these methodologies independently in their own laboratories and, over time, increase our understanding of RGS function in vivo.
Gold
Stephen J
SJ
Department of Psychiatry, University of Texas Southwestern Medical Center, Dallas 75390, USA.
Zachariou
Venetia
V
D000818:Animals; D001007:Anxiety / Q000139:chemically induced; D001522:Behavior, Animal / Q000187:drug effects*; D001921:Brain / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D005786:Gene Expression Regulation; D013287:Illicit Drugs / Q000378:metabolism*; D017403:In Situ Hybridization; D007858:Learning / Q000187:drug effects; D051379:Mice; D008817:Mice, Mutant Strains; D008822:Mice, Transgenic; D009020:Morphine / Q000378:metabolism; D009043:Motor Activity / Q000187:drug effects; D010641:Phenotype; D020710:RGS Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis*
D013287:Illicit Drugs; D020710:RGS Proteins; D012333:RNA, Messenger; D009020:Morphine