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Despite substantial degradation, 2‐arachidonoylglycerol is a potent full efficacy agonist mediating CB 1 receptor‐dependent G‐protein activation in rat cerebellar membranes

Despite substantial degradation, 2‐arachidonoylglycerol is a potent full efficacy agonist... Two endocannabinoids, arachidonoyl ethanolamide (AEA) and 2‐arachidonoylglycerol (2‐AG) bind and activate G‐protein‐coupled cannabinoid receptors, but limited data exist on their relative ability to activate G‐proteins. Here we assess agonist potency and efficacy of various cannabinoids, including 2‐AG, HU‐310 (2‐arachidonoyl glyceryl ether, a third putative endocannabinoid), HU‐313 (another ether analogue of 2‐AG), AEA, R‐methanandamide (an enzymatically stable analogue of AEA), and CP‐55,940 at rat brain CB1 receptors using agonist‐stimulated (35S)‐GTPγS binding to cerebellar membranes and whole brain sections. Degradation of endocannabinoids under experimental conditions was monitored by HPLC. To enhance efficacy differences, agonist dose‐response curves were generated using increasing GDP concentrations. At 10−6 M GDP, all compounds, except HU‐313, produced full agonists responses ∼2.5 fold over basal. The superior efficacy of 2‐AG over all other compounds became evident by increasing GDP (10−5 and 10−4 M). In membrane incubations, 2‐AG was degraded by 85% whereas AEA and HU‐310 were stable. Pretreatment of membranes with phenylmethylsulphonyl fluoride inhibited 2‐AG degradation, resulting in 2 fold increase in agonist potency. Such pretreatment had no effect on AEA potency. Responses in brain sections were otherwise consistent with membrane binding data, but 2‐AG evoked only a weak signal in brain sections, apparently due to more extensive degradation. These data establish that even under conditions of substantial degradation, 2‐AG is a full efficacy agonist, clearly more potent than AEA, in mediating CB1 receptor‐dependent G‐protein activity in native membranes. British Journal of Pharmacology (2001) 134, 664–672; doi:10.1038/sj.bjp.0704297 http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png British Journal of Pharmacology Wiley

Despite substantial degradation, 2‐arachidonoylglycerol is a potent full efficacy agonist mediating CB 1 receptor‐dependent G‐protein activation in rat cerebellar membranes

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Publisher
Wiley
Copyright
2001 British Pharmacological Society
ISSN
0007-1188
eISSN
1476-5381
DOI
10.1038/sj.bjp.0704297
pmid
11588122
Publisher site
See Article on Publisher Site

Abstract

Two endocannabinoids, arachidonoyl ethanolamide (AEA) and 2‐arachidonoylglycerol (2‐AG) bind and activate G‐protein‐coupled cannabinoid receptors, but limited data exist on their relative ability to activate G‐proteins. Here we assess agonist potency and efficacy of various cannabinoids, including 2‐AG, HU‐310 (2‐arachidonoyl glyceryl ether, a third putative endocannabinoid), HU‐313 (another ether analogue of 2‐AG), AEA, R‐methanandamide (an enzymatically stable analogue of AEA), and CP‐55,940 at rat brain CB1 receptors using agonist‐stimulated (35S)‐GTPγS binding to cerebellar membranes and whole brain sections. Degradation of endocannabinoids under experimental conditions was monitored by HPLC. To enhance efficacy differences, agonist dose‐response curves were generated using increasing GDP concentrations. At 10−6 M GDP, all compounds, except HU‐313, produced full agonists responses ∼2.5 fold over basal. The superior efficacy of 2‐AG over all other compounds became evident by increasing GDP (10−5 and 10−4 M). In membrane incubations, 2‐AG was degraded by 85% whereas AEA and HU‐310 were stable. Pretreatment of membranes with phenylmethylsulphonyl fluoride inhibited 2‐AG degradation, resulting in 2 fold increase in agonist potency. Such pretreatment had no effect on AEA potency. Responses in brain sections were otherwise consistent with membrane binding data, but 2‐AG evoked only a weak signal in brain sections, apparently due to more extensive degradation. These data establish that even under conditions of substantial degradation, 2‐AG is a full efficacy agonist, clearly more potent than AEA, in mediating CB1 receptor‐dependent G‐protein activity in native membranes. British Journal of Pharmacology (2001) 134, 664–672; doi:10.1038/sj.bjp.0704297

Journal

British Journal of Pharmacology – Wiley

Published: Oct 1, 2001

References

  • Biosynthesis, release and degradation of the novel endogenous cannabimimetic metabolite 2‐arachidonoylglycerol in mouse neuroblastoma cells
    BISOGNO, BISOGNO; SEPE, SEPE; MELCK, MELCK; MAURELLI, MAURELLI; DE PETROCELLIS, DE PETROCELLIS; DI MARZO, DI MARZO
  • A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein‐dye binding
    BRADFORD, BRADFORD
  • Cannabinoid agonist signal transduction in rat brain: comparison of cannabinoid agonists in receptor binding, G‐protein activation, and adenylyl cyclase inhibition
    BREIVOGEL, BREIVOGEL; CHILDERS, CHILDERS
  • Cannabinoid receptor agonist efficacy for stimulating ( 35 S)‐GTPγS binding to rat cerebellar membranes correlates with agonist‐induced decreases in GDP affinity
    BREIVOGEL, BREIVOGEL; SELLEY, SELLEY; CHILDERS, CHILDERS
  • Isolation and structure of a brain constituent that binds to the cannabinoid receptor
    DEVANE, DEVANE; HANUS, HANUS; BREUER, BREUER; PERTWEE, PERTWEE; STEVENSON, STEVENSON; GRIFFIN, GRIFFIN; GIBSON, GIBSON; MANDELBAUM, MANDELBAUM; ETINGER, ETINGER; MECHOULAM, MECHOULAM
  • Levels, metabolism, and pharmacological activity of anandamide in CB 1 cannabinoid receptor knockout mice: evidence for non‐CB 1 , non‐CB 2 receptor‐mediated actions of anandamide in mouse brain
    DI MARZO, DI MARZO; BREIVOGEL, BREIVOGEL; TAO, TAO; BRIDGEN, BRIDGEN; RAZDAN, RAZDAN; ZIMMER, ZIMMER; ZIMMER, ZIMMER; MARTIN, MARTIN
  • Molecular characterization of a rat brain monoacylglycerol lipase that participates in the hydrolysis of the endocannabinoid 2‐arachidonylglycerol
    DINH, DINH; LESLIE, LESLIE; ZHOU, ZHOU; PIOMELLI, PIOMELLI
  • Agonist selective regulation of G‐proteins by cannabinoid CB 1 and CB 2 receptors
    GLASS, GLASS; NORTHUP, NORTHUP
  • Endocannabinoid 2‐arachidonyl glycerol is a full agonist through human type 2 cannabinoid receptor: antagonism by anandamide
    GONSIOREK, GONSIOREK; LUNN, LUNN; FAN, FAN; NARULA, NARULA; LUNDELL, LUNDELL; HIPKIN, HIPKIN
  • Enzymes of porcine brain hydrolysing 2‐arachidonoylglycerol, an endogenous ligand of cannabinoid receptors
    GOPARAJU, GOPARAJU; UEDA, UEDA; TANIGUCHI, TANIGUCHI; YAMAMOTO, YAMAMOTO
  • Anandamide aminohydrolase reacting with 2‐arachidonoylglycerol, another cannabinoid ligand
    GOPARAJU, GOPARAJU; UEDA, UEDA; YAMAQUCHI, YAMAQUCHI; YAMAMOTO, YAMAMOTO
  • Evaluation of cannabinoid receptor agonists and antagonists using the guanosine‐5′‐O‐(3‐( 35 S)thio)‐triphosphate binding assay in rat cerebellar membranes
    GRIFFIN, GRIFFIN; ATKINSON, ATKINSON; SHOWALTER, SHOWALTER; MARTIN, MARTIN; ABOOD, ABOOD
  • 2‐Arachidonyl glyceryl ether, an endogenous agonist of the cannabinoid CB 1 receptor
    HANUS, HANUS; ABU‐LAFI, ABU‐LAFI; FRIDE, FRIDE; BREUER, BREUER; VOGEL, VOGEL; SHALEV, SHALEV; KUSTANOVICH, KUSTANOVICH; MECHOULAM, MECHOULAM
  • Characterization and localization of cannabinoid receptors in rat brain: a quantitative in vitro autoradiographic study
    HERKENHAM, HERKENHAM; LYNN, LYNN; JOHNSON, JOHNSON; MELVIN, MELVIN; DE COSTA, DE COSTA; RICE, RICE
  • Biochemistry and pharmacology of the endocannabinoids arachidonylethanolamide and 2‐arachidonylglycerol
    HILLARD, HILLARD
  • Oxygenation of the endocannabinoid, 2‐arachidonylglycerol, to glyceryl prostaglandins by cyclooxygenase‐2
    KOZAK, KOZAK; ROWLINSON, ROWLINSON; MARNETT, MARNETT
  • (γ‐ 35 S)‐GTP autoradiography allows region‐specific detection of muscarinic receptor‐dependent G protein activation in the chick optic tectum
    KURKINEN, KURKINEN; KOISTINAHO, KOISTINAHO; LAITINEN, LAITINEN
  • Selective detection of adenosine A 1 receptor‐dependent G protein activity in basal and stimulated conditions of rat brain ( 35 S)‐guanosine 5′(γ‐thio)triphosphate autoradiography
    LAITINEN, LAITINEN
  • Measurement of guanine nucleotide‐binding protein activation by A 1 adenosine receptor agonists in bovine brain membranes: stimulation of guanosine‐5′‐ O ‐(3‐( 35 S)thio)triphosphate binding
    LORENZEN, LORENZEN; FUSS, FUSS; VOGT, VOGT; SCHWABE, SCHWABE
  • Interaction of full and partial agonists of the A 1 adenosine receptor with receptor/G protein complexes in rat brain membranes
    LORENZEN, LORENZEN; GUERRA, GUERRA; VOGT, VOGT; SCHWABE, SCHWABE
  • Activation of various subtypes of G‐protein α subunits by partial agonists of the adenosine A 1 receptor
    LORENZEN, LORENZEN; LANG, LANG; SCHWABE, SCHWABE
  • Structure of a cannabinoid receptor and functional expression of the cloned cDNA
    MATSUDA, MATSUDA; LOLAIT, LOLAIT; BROWNSTEIN, BROWNSTEIN; YOUNG, YOUNG; BONNER, BONNER
  • Identification of an endogenous 2‐monoglyceride, present in canine cut, that binds to cannabinoid receptors
    MECHOULAM, MECHOULAM; BEN‐SHABAT, BEN‐SHABAT; HANUS, HANUS; LIGUMSKY, LIGUMSKY; KAMINSKI, KAMINSKI; SCHATZ, SCHATZ; GOPHER, GOPHER; ALMOG, ALMOG; MARTIN, MARTIN; COMPTON, COMPTON; PERTWEE, PERTWEE; GRIFFIN, GRIFFIN; BAYEWHICH, BAYEWHICH; BARG, BARG; VOGEL, VOGEL
  • CB 1 receptor‐G protein association. Subtype selectivity is determined by distinct intracellular domains
    MUKHOPADHYAY, MUKHOPADHYAY; HOWLETT, HOWLETT
  • The CB 1 cannabinoid receptor juxtamembrane C‐terminal peptide confers activation to specific G proteins in brain
    MUKHOPADHYAY, MUKHOPADHYAY; MCINTOSH, MCINTOSH; HOUSTON, HOUSTON; HOWLETT, HOWLETT
  • Molecular characterization of a peripheral receptor for cannabinoids
    MUNRO, MUNRO; THOMAS, THOMAS; ABU‐SHAAR, ABU‐SHAAR
  • Interaction of 5‐HT 1B/D ligands with recombinant activity and modulation by G‐protein activation state
    PAUWELS, PAUWELS; PALMIER, PALMIER; DUPUIS, DUPUIS; COLBAERT, COLBAERT
  • Neuropharmacology and therapeutic potential of cannabinoids
    PERTWEE, PERTWEE
  • Activation of cannabinoid receptors in rat brain by WIN 55212‐2 produces coupling to multiple G protein α‐subunits with different potencies
    PRATHER, PRATHER; MARTIN, MARTIN; BREIVOGEL, BREIVOGEL; CHILDERS, CHILDERS
  • μ‐Opioid receptor‐stimulated guanosine‐5′‐ O ‐(γ‐thio)‐triphosphate binding in rat thalamus and cultured cell lines: signal transduction mechanisms underlying agonist efficacy
    SELLEY, SELLEY; SIM, SIM; XIAO, XIAO; LIU, LIU; CHILDERS, CHILDERS
  • Cannabinoid receptor stimulation of guanosine‐5′‐ O ‐(3‐( 35 S)‐thio)triphosphate binding in rat brain membranes
    SELLEY, SELLEY; STARK, STARK; SIM, SIM; XIAO, XIAO; CHILDERS, CHILDERS
  • Effects of cronic treatment with Δ 9 ‐tetrahydrocannabinol on cannabinoid‐stimulated ( 35 S)GTPγS autoradiography in rat brain
    SIM, SIM; HAMPSON, HAMPSON; DEADWYLER, DEADWYLER; CHILDERS, CHILDERS
  • A second endogenous cannabinoid that modulates long‐term potentiation
    STELLA, STELLA; SCHWEITZER, SCHWEITZER; PIOMELLI, PIOMELLI
  • 2‐arachidonoylglycerol: A possible endogenous cannabinoid receptor ligand in brain
    SUGIURA, SUGIURA; KONDO, KONDO; SUKAGAWA, SUKAGAWA; NAKANE, NAKANE; SHINODA, SHINODA; ITOH, ITOH; YAMASHITA, YAMASHITA; WAKU, WAKU
  • Evidence that the cannabinoid CB 1 receptor is a 2‐arachidonoylglycerol receptor
    SUGIURA, SUGIURA; KODAKA, KODAKA; NAKANE, NAKANE; MIYASHITA, MIYASHITA; KONDO, KONDO; SUHARA, SUHARA; TAKAYAMA, TAKAYAMA; WAKU, WAKU; SEKI, SEKI; BABA, BABA; ISHIMA, ISHIMA
  • Evidence that 2‐arachidonoylglycerol but not N‐palmitoylethanolamine or anandamide is the physiological ligand for the cannabinoid CB 2 receptor
    SUGIURA, SUGIURA; KONDO, KONDO; KISHIMOTO, KISHIMOTO; MIYASHITA, MIYASHITA; NAKANE, NAKANE; KODAKA, KODAKA; SUHARA, SUHARA; TAKAYAMA, TAKAYAMA; WAKU, WAKU
  • Influence of G protein type on agonist efficacy
    YANG, YANG; LANIER, LANIER

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