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Cannabinoid receptor stimulation of guanosine-5'-O-(3-[35S]thio)triphosphate binding in rat brain membranes.
Life Sci. 1996; 59(8):659-68.LS

Abstract

Cannabinoid receptors belong to the class of G-protein-coupled receptors which inhibit adenylyl cyclase. Coupling of receptors to G-proteins can be assessed by the ability of agonists to stimulate guanosine-5'-O-(3-[35S]thio)triphosphate ([35S]GTP gamma S) binding in the presence of excess GDP. The present study examined the effect of cannabinoid agonists on [35S]GTP gamma S binding in rat brain membranes. Assays were conducted with 0.05 nM [35S]GTP gamma S, incubated with rat cerebellar membranes, 1-30 microM GDP and the cannabinoid agonist WIN 55212-2. Results showed that the ability of WIN 55212-2 to stimulate [35S]GTP gamma S binding increased with increasing concentrations of GDP, with 10-30 microM GDP providing approximately 150-200% stimulation by the cannabinoid agonist. The pharmacology of cannabinoid agonist stimulation of [35S]GTP gamma S binding paralleled that of previously reported receptor binding and adenylyl cyclase assays, and agonist stimulation of [35S]GTP gamma S binding was blocked by the cannabinoid antagonist SR141716A. Brain regional studies revealed widespread stimulation of [35S]GTP gamma S binding by WIN 55212-2 in a number of brain areas, consistent with in vitro [35S]GTP gamma S autoradiography. These results demonstrate that [35S]GTP gamma S binding in the presence of excess GDP is an effective measure of cannabinoid receptor coupling to G-proteins in brain membranes.

Authors+Show Affiliations

Department of Physiology and Pharmacology, Bowman Gray School of Medicine, Winston-Salem, NC 27157, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

8761016

Citation

Selley, D E., et al. "Cannabinoid Receptor Stimulation of guanosine-5'-O-(3-[35S]thio)triphosphate Binding in Rat Brain Membranes." Life Sciences, vol. 59, no. 8, 1996, pp. 659-68.
Selley DE, Stark S, Sim LJ, et al. Cannabinoid receptor stimulation of guanosine-5'-O-(3-[35S]thio)triphosphate binding in rat brain membranes. Life Sci. 1996;59(8):659-68.
Selley, D. E., Stark, S., Sim, L. J., & Childers, S. R. (1996). Cannabinoid receptor stimulation of guanosine-5'-O-(3-[35S]thio)triphosphate binding in rat brain membranes. Life Sciences, 59(8), 659-68.
Selley DE, et al. Cannabinoid Receptor Stimulation of guanosine-5'-O-(3-[35S]thio)triphosphate Binding in Rat Brain Membranes. Life Sci. 1996;59(8):659-68. PubMed PMID: 8761016.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cannabinoid receptor stimulation of guanosine-5'-O-(3-[35S]thio)triphosphate binding in rat brain membranes. AU - Selley,D E, AU - Stark,S, AU - Sim,L J, AU - Childers,S R, PY - 1996/1/1/pubmed PY - 1996/1/1/medline PY - 1996/1/1/entrez SP - 659 EP - 68 JF - Life sciences JO - Life Sci VL - 59 IS - 8 N2 - Cannabinoid receptors belong to the class of G-protein-coupled receptors which inhibit adenylyl cyclase. Coupling of receptors to G-proteins can be assessed by the ability of agonists to stimulate guanosine-5'-O-(3-[35S]thio)triphosphate ([35S]GTP gamma S) binding in the presence of excess GDP. The present study examined the effect of cannabinoid agonists on [35S]GTP gamma S binding in rat brain membranes. Assays were conducted with 0.05 nM [35S]GTP gamma S, incubated with rat cerebellar membranes, 1-30 microM GDP and the cannabinoid agonist WIN 55212-2. Results showed that the ability of WIN 55212-2 to stimulate [35S]GTP gamma S binding increased with increasing concentrations of GDP, with 10-30 microM GDP providing approximately 150-200% stimulation by the cannabinoid agonist. The pharmacology of cannabinoid agonist stimulation of [35S]GTP gamma S binding paralleled that of previously reported receptor binding and adenylyl cyclase assays, and agonist stimulation of [35S]GTP gamma S binding was blocked by the cannabinoid antagonist SR141716A. Brain regional studies revealed widespread stimulation of [35S]GTP gamma S binding by WIN 55212-2 in a number of brain areas, consistent with in vitro [35S]GTP gamma S autoradiography. These results demonstrate that [35S]GTP gamma S binding in the presence of excess GDP is an effective measure of cannabinoid receptor coupling to G-proteins in brain membranes. SN - 0024-3205 UR - https://www.unboundmedicine.com/medline/citation/8761016/Cannabinoid_receptor_stimulation_of_guanosine_5'_O__3_[35S]thio_triphosphate_binding_in_rat_brain_membranes_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0024320596003475 DB - PRIME DP - Unbound Medicine ER -