Pharmacy Faculty Publications

Title

Differential Effects of Opioid Agonists on G-Protein Expression in CHO Cells Expressing the Cloned Human Opioid Receptors

Document Type

Article

Publication Date

9-5-2008

Journal Title

Brain Research Bulletin

Volume

77

Issue

1

First Page

49

Last Page

54

DOI

10.1016/j.brainresbull.2008.05.003

Abstract

Recent evidence indicates that agonist ligands of G protein coupled receptors (GPCR) can activate different signaling systems. Such “agonist-directed” signaling also occurs with opioid receptors. Previous work from our laboratory showed that chronic morphine, but not DAMGO, up-regulates the expression of Gα12 and that both morphine and DAMGO decreased Gαi3 expression in CHO cells expressing the cloned human mu opioid receptor. In this study, we tested the hypothesis that chronic opioid regulation of G protein expression is agonist-directed. Following a 20h treatment of CHO cells expressing the cloned human mu (hMOR-CHO), delta (hDOR-CHO) or kappa (hKOR-CHO) opioid receptors with various opioid agonists, we determined the expression level of Gα12 and Gαi3 by Western blots. Among five mu agonists (morphine, etorphine, DADLE, DAMGO, herkinorin) tested with hMOR-CHO cells, only chronic morphine and etorphine up-regulated Gα12 expression. All five mu agonists decreased Gαi3 expression. Among six delta agonists (SNC80, DPDPE, deltorphin-1, morphine, DADLE, etorphine) tested with hDOR-CHO cells, all six agonists down-regulated Gαi3 expression or moderately up-regulated Gα12 expression. Among five kappa agonists, ((−)-ethylketocyclazocine, salvinorin A, U69,593, etorphine, (−)-U50,488) tested with hKOR-CHO cells, only chronic (−)-U50,488 and (−)-EKC up-regulated Gα12 expression. All kappa agonists decreased Gαi3 expression. These data demonstrate that chronic opioid agonist regulation of G protein expression depends not only on the agonist tested, but also on the type of opioid receptor expressed in a common cellular host, providing additional evidence for agonist-directed signaling.

Keywords

CHO cells, Chinese hamster ovary cells, hMOR-CHO, CHO cells expressing the cloned human & mu, opioid receptor, hDOR-CHO, CHO cells expressing the cloned human & delta, opioid receptor