Coupling of the cloned rat κ-opioid receptor to adenylyl cyclase is dependent on receptor expression

R. A. Hirst, K. Hirota, D. K. Grandy, D. G. Lambert

    Research output: Contribution to journalArticlepeer-review

    12 Scopus citations


    This study describes the coupling of the recombinant rat κ-opioid receptor expressed in Chinese hamster ovary (CHO) cells to adenylyl cyclase and the effects of receptor density. The binding of [3H]diprenorphine ([3H]DPN) was dose dependent and saturable in membranes prepared from cells of early (p4-7) and late (p14-17) passage after transfection. As passage increased the receptor numbers (B(max)) declined from 231 ± 24 (early) to 31 ± 2 fmol/mg protein (late) but the equilibrium dissociation constant (K(d)) did not change. Spiradoline dose dependently displaced [3H]DPN from membranes prepared from early and late cells revealing both high (Ki(H)) and low (Ki(L)) affinity binding sites. There were no significant differences in the proportion of these sites (approximately 50% Ki(L):50% Ki(H)), and whilst spiradoline was generally less potent in late cells the differences were small and failed to reach statistical significance. In contrast, spiradoline produced a dose dependent inhibition of forskolin stimulated cAMP formation in whole cells with pIC50 of 8.62 and 8.00 in early compared with late cells. In addition, the maximum inhibition was dramatically reduced from 47 to 22%. Etorphine, (±)bremazocine, ICI-204,448 and (±)trans-U-50488 methanesulfonate (1 μM), compounds with activity at κ-receptors, produced a greater inhibition of cAMP formation in early (42.2, 45.8, 50.2 and 50.5%, respectively) than late (12.9, 11.8, 13.5 and 7.8%, respectively) cells, indicating that expression dependent inhibition of cAMP formation was not κ- agonist specific. Collectively, these data suggest that in CHO cells, κ- opioid receptor coupling to adenylyl cyclase is dependent on receptor expression levels.

    Original languageEnglish (US)
    Pages (from-to)119-122
    Number of pages4
    JournalNeuroscience Letters
    Issue number2
    StatePublished - Aug 29 1997


    • Adenylyl cyclase
    • Chinese hamster ovary cells
    • Recombinant κ-opioid receptor
    • Spiradoline
    • cAMP

    ASJC Scopus subject areas

    • General Neuroscience


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