Evidence that 2-arachidonoylglycerol but not N-palmitoylethanolamine or anandamide is the physiological ligand for the cannabinoid CB2 receptor. Comparison of the agonistic activities of various cannabinoid receptor ligands in HL-60 cells

Takayuki Sugiura, Sachiko Kondo, Seishi Kishimoto, Tomoyuki Miyashita, Shinji Nakane, Tomoko Kodaka, Yoshitomo Suhara, Hiroaki Takayama, Keizo Waku

Research output: Contribution to journalArticle

293 Citations (Scopus)

Abstract

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. G(i) or G(o) 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.

Original languageEnglish
Pages (from-to)605-612
Number of pages8
JournalJournal of Biological Chemistry
Volume275
Issue number1
DOIs
Publication statusPublished - 2000 Jan 7
Externally publishedYes

Fingerprint

Cannabinoid Receptor CB2
Cannabinoid Receptors
Ligands
Cannabinoid Receptor CB1
rimonabant
Arachidonic Acid
Cells
Lipoxygenase Inhibitors
Cyclooxygenase Inhibitors
Immune system
Pertussis Toxin
Metabolites
2-arachidonylglycerol
anandamide
palmidrol
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Evidence that 2-arachidonoylglycerol but not N-palmitoylethanolamine or anandamide is the physiological ligand for the cannabinoid CB2 receptor. Comparison of the agonistic activities of various cannabinoid receptor ligands in HL-60 cells. / Sugiura, Takayuki; Kondo, Sachiko; Kishimoto, Seishi; Miyashita, Tomoyuki; Nakane, Shinji; Kodaka, Tomoko; Suhara, Yoshitomo; Takayama, Hiroaki; Waku, Keizo.

In: Journal of Biological Chemistry, Vol. 275, No. 1, 07.01.2000, p. 605-612.

Research output: Contribution to journalArticle

@article{2eeb6e4686b445f4ad3b861e90abba07,
title = "Evidence that 2-arachidonoylglycerol but not N-palmitoylethanolamine or anandamide is the physiological ligand for the cannabinoid CB2 receptor. Comparison of the agonistic activities of various cannabinoid receptor ligands in HL-60 cells",
abstract = "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. G(i) or G(o) 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.",
author = "Takayuki Sugiura and Sachiko Kondo and Seishi Kishimoto and Tomoyuki Miyashita and Shinji Nakane and Tomoko Kodaka and Yoshitomo Suhara and Hiroaki Takayama and Keizo Waku",
year = "2000",
month = "1",
day = "7",
doi = "10.1074/jbc.275.1.605",
language = "English",
volume = "275",
pages = "605--612",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "1",

}

TY - JOUR

T1 - Evidence that 2-arachidonoylglycerol but not N-palmitoylethanolamine or anandamide is the physiological ligand for the cannabinoid CB2 receptor. Comparison of the agonistic activities of various cannabinoid receptor ligands in HL-60 cells

AU - Sugiura, Takayuki

AU - Kondo, Sachiko

AU - Kishimoto, Seishi

AU - Miyashita, Tomoyuki

AU - Nakane, Shinji

AU - Kodaka, Tomoko

AU - Suhara, Yoshitomo

AU - Takayama, Hiroaki

AU - Waku, Keizo

PY - 2000/1/7

Y1 - 2000/1/7

N2 - 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. G(i) or G(o) 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.

AB - 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. G(i) or G(o) 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.

UR - http://www.scopus.com/inward/record.url?scp=0034614374&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034614374&partnerID=8YFLogxK

U2 - 10.1074/jbc.275.1.605

DO - 10.1074/jbc.275.1.605

M3 - Article

C2 - 10617657

AN - SCOPUS:0034614374

VL - 275

SP - 605

EP - 612

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 1

ER -