Function and Regulation of Somatostatin Receptor Subtypes

The five known somatostatin receptors serve unique biological roles by virtue of their tissue-specific expression and particular biochemical properties. However, the function of any individual receptor in its normal physiological milieu is not understood. Studies to address this problem have been difficult because tissues and cell lines often express multiple somatostatin receptors and, in the absence of receptor-selective somatostatin analogues, the actions of individual receptors cannot be identified. Moreover, the biological and biochemical actions of somatostatin receptors depend on their cellular environment, so that the behaviour of a receptor expressed in heterologous cells does not necessarily mimic that of endogenous receptors. We have developed two approaches to examine somatostatin receptors which circumvent these problems. Using a biotinylated somatostatin analogue for affinity purification, we isolated somatostatin receptors together with associated G proteins. Subsequent analysis of the purified complex with G protein-specific antibodies showed that the somatostatin receptors in AR42J cells preferentially couple with two pertussis toxin-sensitive G proteins: Gi alpha 1 and Gi alpha 3. To examine individual receptor types, we developed receptor-specific antibodies and used them to show that both sstr1 and sstr2 proteins were present in the GH4C1 pituitary cell line whereas AR42J cells contained sstr2 but not sstr1. Immunoprecipitation of receptor-G protein complexes with GH4C1 cells showed that sstr1 and sstr2 are both coupled to pertussis toxin-sensitive G proteins, in contrast to the results observed when these receptors are overexpressed in some non-endocrine cells. We also showed that the somatostatin receptors in GH4C1 cells are subject to both homologous and heterologous hormonal regulation. The mechanisms involved in the regulation of different receptor types are now being characterized using the receptor-specific antibodies to isolate the individual receptor proteins. Elucidating signal transduction by endogenous somatostatin receptors as well as their hormonal regulation will be critical for understanding the functions of these receptors in the different physiological targets of somatostatin.