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J Biol Chem, Vol. 275, Issue 1, 605-612, January 7, 2000
,
From the Faculty of Pharmaceutical Sciences, Teikyo University,
Sagamiko, Tsukui-gun, Kanagawa 199-0195, Japan
We examined the effect of 2-arachidonoylglycerol,
an endogenous cannabinoid receptor ligand, on the intracellular free
Ca2+ concentrations in HL-60 cells that express the
cannabinoid CB2 receptor. We found that 2-arachidonoylglycerol induces
a rapid transient increase in intracellular free Ca2+
concentrations in HL-60 cells. The response was affected by neither cyclooxygenase inhibitors nor lipoxygenase inhibitors, suggesting that
arachidonic acid metabolites are not involved. Consistent with this
notion, free arachidonic acid was devoid of any agonistic activity.
Importantly, the Ca2+ transient induced by
2-arachidonoylglycerol was blocked by pretreatment of the cells with
SR144528, a CB2 receptor-specific antagonist, but not with SR141716A, a
CB1 receptor-specific antagonist, indicating the involvement of the CB2
receptor but not the CB1 receptor in this cellular response.
Gi or Go is also assumed to be involved, because pertussis toxin treatment of the cells abolished the response. We further examined the structure-activity relationship. We found that
2-arachidonoylglycerol is the most potent compound among a number of
naturally occurring cannabimimetic molecules. Interestingly, anandamide
and N-palmitoylethanolamine, other putative endogenous ligands, were found to be a weak partial agonist and an inactive ligand, respectively. These results strongly suggest that the CB2
receptor is originally a 2-arachidonoylglycerol receptor, and
2-arachidonoylglycerol is the intrinsic natural ligand for the CB2
receptor that is abundant in the immune system.
To whom correspondence should be addressed. Fax:
81-426- 85-1345.
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