Regulation of Fas receptor/Fas-associated protein with death domain apoptotic complex and associated signalling systems by cannabinoid receptors in the mouse brain.
OBJECTIVE
Natural and synthetic cannabinoids (CBs) induce deleterious or beneficial actions on neuronal survival. The Fas-associated protein with death domain (FADD) promotes apoptosis, and its phosphorylated form (p-FADD) mediates non-apoptotic actions. The regulation of Fas/FADD, mitochondrial apoptotic proteins and other pathways by CB receptors was investigated in the mouse brain.


METHODS
Wild-type, CB(1) and CB(2) receptor knock-out (KO) mice were used to assess differences in receptor genotypes. CD1 mice were used to evaluate the effects of CB drugs on canonical apoptotic pathways and associated signalling systems. Target proteins were quantified by Western blot analysis.


RESULTS
In brain regions of CB(1) receptor KO mice, Fas/FADD was reduced, but p-Ser191 FADD and the p-FADD/FADD ratio were increased. In CB(2) receptor KO mice, Fas/FADD was increased, but the p-FADD/FADD ratio was not modified. In mutant mice, cleavage of poly(ADP-ribose)-polymerase (PARP) did not indicate alterations in brain cell death. In CD1 mice, acute WIN55212-2 (CB(1) receptor agonist), but not JWH133 (CB(2) receptor agonist), inversely modulated brain FADD and p-FADD. Chronic WIN55212-2 induced FADD down-regulation and p-FADD up-regulation. Acute and chronic WIN55212-2 did not alter mitochondrial proteins or PARP cleavage. Acute, but not chronic, WIN55212-2 stimulated activation of anti-apoptotic (ERK, Akt) and pro-apoptotic (JNK, p38 kinase) pathways.


CONCLUSIONS
CB(1) receptors appear to exert a modest tonic activation of Fas/FADD complexes in brain. However, chronic CB(1) receptor stimulation decreased pro-apoptotic FADD and increased non-apoptotic p-FADD. The multifunctional protein FADD could participate in the mechanisms of neuroprotection induced by CBs.
Alvaro-Bartolomé
M
M
Laboratori de Neurofarmacologia, IUNICS, Universitat de les Illes Balears, Palma de Mallorca, Spain.
Esteban
S
S
García-Gutiérrez
M S
MS
Manzanares
J
J
Valverde
O
O
García-Sevilla
J A
JA
D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D048588:Benzoxazines / Q000494:pharmacology; D001921:Brain / Q000187:drug effects / Q000378:metabolism*; D002186:Cannabinoids / Q000494:pharmacology; D016923:Cell Death / Q000235:genetics; D015536:Down-Regulation; D053200:Fas-Associated Death Domain Protein / Q000378:metabolism*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009025:Morpholines / Q000494:pharmacology; D009281:Naphthalenes / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D043884:Receptor, Cannabinoid, CB1 / Q000819:agonists* / Q000235:genetics; D043885:Receptor, Cannabinoid, CB2 / Q000819:agonists* / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects*; D015854:Up-Regulation; D019014:fas Receptor / Q000378:metabolism*
D051017:Apoptosis Regulatory Proteins; D048588:Benzoxazines; D002186:Cannabinoids; D053200:Fas-Associated Death Domain Protein; D009025:Morpholines; D009281:Naphthalenes; D043884:Receptor, Cannabinoid, CB1; D043885:Receptor, Cannabinoid, CB2; D019014:fas Receptor; C070417:(3R)-((2,3-dihydro-5-methyl-3-((4-morpholinyl)methyl)pyrrolo-(1,2,3-de)-1,4-benzoxazin-6-yl)(1-naphthalenyl))methanone; D011065:Poly(ADP-ribose) Polymerases; C432747:1,1-dimethylbutyl-1-deoxy-Delta(9)-THC