μ-Opioid receptor ligands lack receptor subtype selectivity in the aequorin luminescence-based calcium assay

Abstract

The aim of the present study was to characterize the binding selectivity of the μ-opioid receptor ligands, endomorphin-1, endomorphin-2, and DAMGO, in the in vitro functional assay, based on the changes in intracellular calcium levels. For the experiments Chinese hamster ovary cells, stably expressing human μ-receptor, were used. The μ-agonist-induced calcium responses were significantly inhibited by naloxone, an opioid antagonist with high preference for the μ-opioid receptors. Naloxonazine, a μ1-non-peptide antagonist, inhibited the effect of all tested μ-agonists. However, there was no significant difference in the antagonist effect of naloxonazine on the calcium response induced by μ1- (endomorphin-2) and μ2-agonists (endomorphin-1, DAMGO). [d-Pro2] endomorphin-1 and [d-Pro2]endomorphin-2, putative peptide μ2- and μ1-antagonists, respectively, which had been shown in vivo to inhibit the antinociception induced by μ-agonists, produced no inhibitory effect in our in vitro experiments. Our results demonstrated that there is only one population of the μ-opioid receptors expressed in the Chinese hamster ovary cells. We suggest that the μ-opioid receptors form a homogenous population in the in vitro systems. However, the existence of μ-receptor subtypes in vivo is still pharmacologically possible. © 2007 The Authors.