The role of diacylglycerol lipase in constitutive and angiotensin AT 1 receptor-stimulated cannabinoid CB1 receptor activity

Gábor Turu, Anne Simon, Pál Gyombolai, László Szidonya, G. Bagdy, Zsolt Lenkei, L. Hunyady

Research output: Contribution to journalArticle

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Abstract

The cannabinoid CB1 receptor (CB1R) is a G protein-coupled receptor, which couples to the Gi/o family of heterotrimeric G proteins. The receptor displays both basal and agonist-induced signaling and internalization. Although basal activity of CB1Rs is attributed to constitutive (agonist-independent) receptor activity, studies in neurons suggested a role of postsynaptic endocannabinoid (eCB) release in the persistent activity of presynaptic CB1Rs. To elucidate the role of eCBs in basal CB1R activity, we have investigated the role of diacylglycerol lipase (DAGL) in this process in Chinese hamster ovary (CHO) cells, which are not targeted specifically with eCBs. Agonist-induced G protein activation was determined by detecting dissociation G protein subunits expressed in CHO cells with bioluminescence resonance energy transfer (BRET), after labeling the α and β subunits with Renilla luciferase and enhanced yellow fluorescent protein (EYFP), respectively. Pre-incubation of the cells with tetrahydrolipstatin (THL), a known inhibitor of DAGLs, caused inhibition of the basal activity of CB1R. Moreover, preincubation of CHO and cultured hippocampal neurons with THL increased the number of CB1Rs on the cell membrane, which reflects its inhibitory action on CB1R internalization in non-simulated cells. In CHO cells co-expressing CB1R and angiotensin AT 1 receptors, angiotensin II-induced Go protein activation that was blocked by both a CB1R antagonist and THL. These data indicate that cell-derived eCB mediators have a general role in the basal activity of CB1Rs in non-neural cells and neurons, and that this mechanism can be stimulated by AT1 receptor activation.

Original languageEnglish
Pages (from-to)7753-7757
Number of pages5
JournalJournal of Biological Chemistry
Volume282
Issue number11
DOIs
Publication statusPublished - Mar 16 2007

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Cannabinoid Receptor CB1
Lipoprotein Lipase
Angiotensins
Cricetulus
Ovary
Neurons
Endocannabinoids
Chemical activation
GTP-Binding Proteins
Renilla Luciferases
Cells
Bioluminescence
Heterotrimeric GTP-Binding Proteins
Angiotensin Receptors
Protein Subunits
Cell membranes
G-Protein-Coupled Receptors
Energy Transfer
Energy transfer
Labeling

ASJC Scopus subject areas

  • Biochemistry

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The role of diacylglycerol lipase in constitutive and angiotensin AT 1 receptor-stimulated cannabinoid CB1 receptor activity. / Turu, Gábor; Simon, Anne; Gyombolai, Pál; Szidonya, László; Bagdy, G.; Lenkei, Zsolt; Hunyady, L.

In: Journal of Biological Chemistry, Vol. 282, No. 11, 16.03.2007, p. 7753-7757.

Research output: Contribution to journalArticle

Turu, Gábor ; Simon, Anne ; Gyombolai, Pál ; Szidonya, László ; Bagdy, G. ; Lenkei, Zsolt ; Hunyady, L. / The role of diacylglycerol lipase in constitutive and angiotensin AT 1 receptor-stimulated cannabinoid CB1 receptor activity. In: Journal of Biological Chemistry. 2007 ; Vol. 282, No. 11. pp. 7753-7757.
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