Faculty Collaboration Database

Cannabinoid-receptor-independent cell signalling by N-acylethanolamines. Biochem J 2001 Nov 15;360(Pt 1):67-75

Date

11/07/2001

Pubmed ID

11695993

Pubmed Central ID

PMC1222203

DOI

10.1042/0264-6021:3600067

Scopus ID

2-s2.0-0035890677 (requires institutional sign-in at Scopus site)   71 Citations

Abstract

Anandamide and other polyunsaturated N-acylethanolamines (NAEs) exert biological activity by binding to cannabinoid receptors. These receptors are linked to G(i/o) proteins and their activation leads to extracellular-signal-regulated protein kinase (ERK) and c-Jun N-terminal kinase (JNK) mitogen-activated protein kinase (MAP kinase) activation, inhibition of cAMP-dependent signalling and complex changes in the expression of various genes. Saturated and monounsaturated NAEs cannot bind to cannabinoid receptors and may thus mediate cell signalling through other targets. Here we report that both saturated/monounsaturated NAEs and anandamide (20:4(n-6) NAE) stimulate cannabinoid-receptor-independent ERK phosphorylation and activator protein-1 (AP-1)-dependent transcriptional activity in mouse epidermal JB6 cells. Using a clone of JB6 P(+) cells with an AP-1 collagen-luciferase reporter construct, we found that 16:0, 18:1(n-9), 18:1(n-7), 18:2(n-6) and 20:4(n-6) NAEs stimulated AP-1-dependent transcriptional activity up to 2-fold, with maximal stimulation at approx. 10-15 microM. Higher NAE concentrations had toxic effects mediated by alterations in mitochondrial energy metabolism. The AP-1 stimulation appeared to be mediated by ERK but not JNK or p38 signalling pathways, because all NAEs stimulated ERK1/ERK2 phosphorylation without having any effect on JNK or p38 kinases. Also, overexpression of dominant negative ERK1/ERK2 kinases completely abolished NAE-induced AP-1 activation. In contrast with 18:1(n-9) NAE and anandamide, the cannabinoid receptor agonist WIN 55,212-2 did not stimulate AP-1 activity and inhibited ERK phosphorylation. The NAE-mediated effects were not attenuated by pertussis toxin and appeared to be NAE-specific, as a close structural analogue, oleyl alcohol, failed to induce ERK phosphorylation. The data support our hypothesis that the major saturated and monounsaturated NAEs are signalling molecules acting through intracellular targets without participation of cannabinoid receptors.

Author List

Berdyshev EV, Schmid PC, Krebsbach RJ, Hillard CJ, Huang C, Chen N, Dong Z, Schmid HH

Author

Cecilia J. Hillard PhD Associate Dean, Center Director, Professor in the Pharmacology and Toxicology department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Analgesics
Animals
Benzoxazines
Cell Line
Cells, Cultured
Dose-Response Relationship, Drug
Epidermis
Ethanolamines
MAP Kinase Signaling System
Mice
Microscopy, Phase-Contrast
Mitochondria
Mitogen-Activated Protein Kinase 8
Mitogen-Activated Protein Kinases
Morpholines
Naphthalenes
Phosphorylation
Radioligand Assay
Receptors, Cannabinoid
Receptors, Drug
Signal Transduction
Time Factors
Transcription Factor AP-1
Transcription, Genetic
p38 Mitogen-Activated Protein Kinases