Cannabinoid receptor 1 ligands revisited: Pharmacological assessment in the ACTOne system

Presley, Chaela S.; Abidi, Ammaar H.; Moore, Bob M.

doi:10.1016/j.ab.2015.12.019

Cited by 10 publications

(9 citation statements)

References 89 publications

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“…In our experimental approach, and in accordance with other studies (Glass and Northup, 1999; Presley et al, 2016), Δ 9 -THC, WIN55212-2 and ACEA significantly stimulated Gα o . Results obtained in knockout animals show that the stimulation of Gα o in mouse cortex is mediated, at least in part, by CB 2 receptors, suggesting a necessary role of this receptor in the cannabinoid-induced activation of Gα o .…”

Section: Discussionsupporting

confidence: 91%

“…There are contradictory results about the ability of cannabinoid drugs to activate Gα s proteins. In this way, there are data from both CHO cell lines (Rinaldi-Carmona et al, 1996; Bonhaus et al, 1998) and HEK cells (Presley et al, 2016) expressing CB 1 receptor, showing the absence of effect as well as a modest but significant coupling of CB 1 to Gα s triggered by different cannabinoids. It has been proposed that ACEA may elevate cAMP through a non-CB 1 mechanism, since there is an increase in cAMP in both cells transfected and non-transfected with CB 1 and pretreated with pertussis toxin (Presley et al, 2016).…”

Section: Discussionmentioning

confidence: 98%

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Biased Agonism of Three Different Cannabinoid Receptor Agonists in Mouse Brain Cortex

et al. 2016

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Cannabinoid receptors are able to couple to different families of G proteins when activated by an agonist drug. It has been suggested that different intracellular responses may be activated depending on the ligand. The goal of the present study was to characterize the pattern of G protein subunit stimulation triggered by three different cannabinoid ligands, Δ9-THC, WIN55212-2, and ACEA in mouse brain cortex. Stimulation of the [35S]GTPγS binding coupled to specific immunoprecipitation with antibodies against different subtypes of G proteins (Gαi1, Gαi2, Gαi3, Gαo, Gαz, Gαs, Gαq/11, and Gα12/13), in the presence of Δ9-THC, WIN55212-2 and ACEA (submaximal concentration 10 μM) was determined by scintillation proximity assay (SPA) technique in mouse cortex of wild type, CB1 knock-out, CB2 knock-out and CB1/CB2 double knock-out mice. Results show that, in mouse brain cortex, cannabinoid agonists are able to significantly stimulate not only the classical inhibitory Gαi/o subunits but also other G subunits like Gαz, Gαq/11, and Gα12/13. Moreover, the specific pattern of G protein subunit activation is different depending on the ligand. In conclusion, our results demonstrate that, in mice brain native tissue, different exogenous cannabinoid ligands are able to selectively activate different inhibitory and non-inhibitory Gα protein subtypes, through the activation of CB1 and/or CB2 receptors. Results of the present study may help to understand the specific molecular pathways involved in the pharmacological effects of cannabinoid-derived drugs.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 98%

Biased Agonism of Three Different Cannabinoid Receptor Agonists in Mouse Brain Cortex

et al. 2016

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“…There have been several reports on the relationship between in vivo psychoactivity and cannabinoid receptor binding affinity (26–28). With the exception of the study by Presley et al .…”

Section: Discussionmentioning

confidence: 99%

“…With the exception of the study by Presley et al . (26), these studies used radio-ligands for measuring receptor binding affinities. Wiley et al .…”

Section: Discussionmentioning

confidence: 99%

Receptor Binding Affinities of Synthetic Cannabinoids Determined by Non-Isotopic Receptor Binding Assay

et al. 2019

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A major predictor of the efficacy of natural or synthetic cannabinoids is their binding affinity to the cannabinoid type I receptor (CB1) in the central nervous system, as the main psychological effects of cannabinoids are achieved via binding to this receptor. Conventionally, receptor binding assays have been performed using isotopes, which are inconvenient owing to the effects of radioactivity. In the present study, the binding affinities of five cannabinoids for purified CB1 were measured using a surface plasmon resonance (SPR) technique as a putative non-isotopic receptor binding assay. Results were compared with those of a radio-isotope-labeled receptor binding assay. The representative natural cannabinoid Δ9-tetrahydrocannabinol and four synthetic cannabinoids, JWH-015, JWH-210, RCS-4, and JWH-250, were assessed using both the SPR biosensor assay and the conventional isotopic receptor binding assay. The binding affinities of the test substances to CB1 were determined to be (from highest to lowest) 9.52 × 10−13 M (JWH-210), 6.54 × 10−12 M (JWH-250), 1.56 × 10−11 M (Δ9-tetrahydrocannabinol), 2.75 × 10−11 M (RCS-4), and 6.80 ×10−11 M (JWH-015) using the non-isotopic method. Using the conventional isotopic receptor binding assay, the same order of affinities was observed. In conclusion, our results support the use of kinetic analysis via SPR in place of the isotopic receptor binding assay. To replace the receptor binding affinity assay with SPR techniques in routine assays, further studies for method validation will be needed in the future.

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“… Normalized signal cAMP driven fluorescence of CP 55,490 in ACTOne CB1 cells from passage 3 to 36. N =26 from all control runs in [2] with EC 50 7.00±1.2 nM, error bars are SEM. …”

Section: Figmentioning

confidence: 99%

Optimization of cAMP fluorescence dataset from ACTOne cannabinoid receptor 1 cell line

2016

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The ACTOne cannabinoid receptor 1 functional system is comprised of transfected HEK cells with the parental cyclic nucleotide gated channel (CNG) co-transfected with cannabinoid receptor 1 (CB1). The ACTOne CB1 cell line was evaluated for cAMP driven fluorescence by optimizing experimental conditions for sensitivity to forskolin and CP 55,940, reading time point, reliability of cell passage number, and pertussis inactivation of Gi/o.

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Cannabinoid receptor 1 ligands revisited: Pharmacological assessment in the ACTOne system

Cited by 10 publications

References 89 publications

Biased Agonism of Three Different Cannabinoid Receptor Agonists in Mouse Brain Cortex

Biased Agonism of Three Different Cannabinoid Receptor Agonists in Mouse Brain Cortex

Receptor Binding Affinities of Synthetic Cannabinoids Determined by Non-Isotopic Receptor Binding Assay

Optimization of cAMP fluorescence dataset from ACTOne cannabinoid receptor 1 cell line

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