Quantitative evaluation of human delta opioid receptor desensitization using the operational model of drug action.

Agonist-mediated desensitization of the opioid receptors is thought to function as a protective mechanism against sustained opioid signaling and therefore may prevent the development of opioid tolerance. However, the exact molecular mechanism of opioid receptor desensitization remains unresolved because of difficulties in measuring and interpreting receptor desensitization. In the present study, we investigated deltorphin II-mediated rapid desensitization of the human delta opioid receptors (hDOR) by measuring guanosine 5'-O-(3-[(35)S]thio)-triphosphate binding and inhibition of cAMP accumulation.

Morphine promotes phosphorylation of the human delta-opioid receptor at serine 363.

After prolonged stimulation, the delta-opioid receptor becomes desensitized by regulatory mechanisms such as receptor phosphorylation, internalization and down-regulation. In this study, we demonstrate that morphine treatment causes phosphorylation of S363 in the C-terminus of the human delta-opioid receptor. Morphine-mediated phosphorylation reached 53+/-8% of maximum deltorphin II-mediated phosphorylation.

Continuous ethanol exposure inhibits agonist-stimulated phosphorylation of p70S6 kinase and ribosomal S6 protein in cultured rat astrocytes.

The effects of ethanol exposure on agonist-stimulated phosphorylation of p70S6 kinase and ribosomal S6 protein were determined in confluent astrocyte monolayers. Basal phosphorylation of p70S6 kinase and S6 protein was either unaffected or reduced, respectively, after exposure to 50 mM ethanol for 4 days. The abilities of norepinephrine, carbachol and epidermal growth factor to phosphorylate these proteins were significantly decreased after ethanol exposure.