Comparative pharmacological properties and functional coupling of mu and delta opioid receptor sites in human neuroblastoma SH-SY5Y cells

Abstract

The characteristics of mu and delta opioid receptor sites present in human neuroblastoma SH-SY5Y cells were investigated using [D-Ala2-N-methyl-Phe4-Gly-(01)5]enkephalin (DAGO) and [2-D-penicillamine, 5-D-penicillamine]enkephalin (DPDPE), which are the most selective radioligands available for mu and delta sites, respectively. Scatchard analysis of the saturation isotherms revealed high affinity binding to a single class of sites for both [3H]DAGO (mu) and [3H]DPDPE (delta). [3H]DAGO labeled twice the number of sites compared to the binding capacity of [3H]DPDPE, yielding a mu/delta ratio of 2:1. Selective suppression of [3H]diprenorphine binding by specific opioid "blocking" ligands also showed a predominance of mu receptors, representing 65-70% of the total opioid sites. Competition binding studies carried out with a series of opiates and opioid peptides displayed higher potencies of mu- and delta-selective ligands in displacing the specific binding of [3H]DAGO and [3H]DPDPE, respectively. The [3H]diprenorphine/agonist competition curves were biphasic, indicating the high and low affinity states of mu and delta receptor sites in SH-SY5Y cells. Guanine nucleotide and sodium had differential effects on the agonist affinity and the proportion of high affinity states of mu and delta receptors. The mu and delta receptor sites were shown to be functionally coupled to adenylate cyclase. All of these data support the independent existence of mu and delta receptor types in human neuroblastoma cells. SH-SY5Y cells, therefore, represent a suitable model for investigating opioid-mediated responses in nerve cell populations.