Reporter gene systems for the study of G protein-coupled receptor signal transduction in mammalian cells

Abstract

Abstract Traditional protocols for the measurement of G protein-coupled receptor (GPCR) signal transduction in mammalian cells have relied upon a number of biochemical techniques. These include the measurement of the rate of guanine nucleotide exchange at the activated G-protein alpha subunit (1), the measurement of the changes in the level of one of a plethora of intracellular second messenger metabolites, such as cAMP, calcium, or inositol phosphates (2), or the activation or inhibition of an ion channel (3). In recent years these assays have been supplemented by the development of reporter gene systems for the study of GPCR signal transduction (4-5). In this chapter we shall describe a number of such reporter gene systems, we shall outline the advantages of reporter genes over standard biochemical analyses, and we shall describe a number of protocols developed for the study of GPCR signal transduction using reporter gene systems.