High content screening to monitor G protein-coupled receptor internalisation.

Abstract

G protein-coupled receptors (GPCRs) fulfil a broad diversity of physiological functions in areas such as neurotransmission, respiration, cardiovascular action, pain and more. Consequently, they are considered as the most successful group of therapeutic targets on the pharmaceutical market, and the search for compounds that interfere with GPCR function in a specific and selective way is a major focus of the pharmaceutical industry. High Content Screening (HCS), a combination of fluorescence microscopic imaging and automated image analysis, has become a frequently employed tool to study test compound effects in cellular disease modelling systems. One way to functionally analyse the effect of test compounds on GPCRs by HCS relies on the broadly observed phenomenon of desensitisation. Agonist stimulation of most GPCRs leads to their intracellular phosphorylation and subsequent internalisation, resulting in the termination of receptor signalling and the seclusion of the GPCR from further extracellular stimulation. Complementary to other functional GPCR drug discovery assays, GPCR internalisation assays enable a desensitisation-focussed pharmacological analysis of test compounds.