Identification of G-protein-coupled receptor mRNA expression by Northern blotting and in situ hybridization.
G-protein-coupled receptor mRNAs are expressed at low levels and therefore present a challenge for the study of their sites and levels of expression. In situ hybridization (ISH) and Northern blotting are powerful methods for the localization of mRNAs and the study of regulation of mRNA expression. ISH combines the power of precise cellular localization with the ability to perform semiquantitative analysis of the mRNA level, whereas Northern blotting has the ability to identify genetic splice variants, or to study multiple RNA molecules sequentially in the same tissue samples. These protocols give step-by-step instructions for the performance of these techniques, and the analysis of the data that can be obtained using them.
Donaldson
Lucy F
LF
Department of Physiology, University of Bristol School of Medical Sciences, United Kingdom.
D000818:Animals; D001345:Autoradiography / Q000379:methods; D015152:Blotting, Northern / Q000379:methods*; D017403:In Situ Hybridization / Q000379:methods*; D015347:RNA Probes; D018075:RNA, Complementary / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis* / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000235:genetics*
D015347:RNA Probes; D018075:RNA, Complementary; D012333:RNA, Messenger; D043562:Receptors, G-Protein-Coupled