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. 2000 Jun;20(3):291–304. doi: 10.1023/A:1007058008477

Opioid and Cannabinoid Receptors Share a Common Pool of GTP-Binding Proteins in Cotransfected Cells, But Not in Cells Which Endogenously Coexpress the Receptors

Ma'anit Shapira 1, Zvi Vogel 2, Yosef Sarne 1,3
PMCID: PMC11537524  PMID: 10789829

Abstract

1. Opioid (μ, δ, κ) and cannabinoid (CB1, CB2) receptors are coupled mainly toGi/Go GTP-binding proteins. The goal of the present study was to determine whether different subtypes of opioid and cannabinoid receptors, when coexpressed in the same cell, share a common reservoir, or utilize different pools, of G proteins.

2. The stimulation of [35S]GTPγS binding by selective opioid and cannabinoid agonists was tested in transiently transfected COS-7 cells, as well as in neuroblastoma cell lines. In COS-7 cells, cotransfection of μ- and δ-opioid receptors led to stimulation of [35S]GTPγS binding by either μ-selective (DAMGO) or δ-selective (DPDPE) agonists. The combined effect of the two agonists was similar to the effect of either DAMGO or DPDPE alone, suggesting the activation of a common G-protein reservoir by the two receptor subtypes.

3. The same phenomenon was observed when COS-7 cells were cotransfected with CB1 cannabinoid receptors and either μ- or δ-opioid receptors.

4. On the other hand, in N18TG2 neuroblastoma cells, which endogenously coexpress CB1 and δ-opioid receptors, as well as in SK-N-SH neuroblastoma cells, which coexpress μ- and δ-opioid receptors, the combined effects of the various agonists (the selective cannabinoid DALN and the selective opioids DPDPE and DAMGO) were additive, implying the activation of different pools of G proteins by each receptorsubtype.

5. These results suggest a fundamental difference between native and artificially transfected cells regarding the compartmentalization of receptors and GTP-binding proteins.

Keywords: opioid receptors, cannabinoid receptors, GTP-binding proteins, cell transfection, [35S]GTPγS binding, neuroblastoma

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