http://www.cnr.it/ontology/cnr/individuo/prodotto/ID196996

Biosynthesis, release and degradation of the novel endogenous cannabimimetic metabolite 2-arachidonoylglycerol in mouse neuroblastoma cells (Articolo in rivista)

Type

Prodotto della ricerca (Classe)
Articolo in rivista (Classe)

Label

Biosynthesis, release and degradation of the novel endogenous cannabimimetic metabolite 2-arachidonoylglycerol in mouse neuroblastoma cells (Articolo in rivista) (literal)

Anno

1997-01-01T00:00:00+01:00 (literal)

Alternative label

Bisogno T, Sepe N, Melck D, Maurelli S, De Petrocellis L, Di Marzo V. (1997)
Biosynthesis, release and degradation of the novel endogenous cannabimimetic metabolite 2-arachidonoylglycerol in mouse neuroblastoma cells
in Biochemical journal (Lond., 1984); PORTLAND PRESS,, LONDON W1N 3AJ ENGLAND (Regno Unito)
(literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori

Bisogno T, Sepe N, Melck D, Maurelli S, De Petrocellis L, Di Marzo V. (literal)

Pagina inizio

671 (literal)

Pagina fine

677 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume

322 (literal)

Rivista

Biochemical journal (Rivista)

Note

ISI Web of Science (WOS) (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni

1. CNR,IST CHIM MOL INTERESSE BIOL, I-80072 ARCO FELICE, NAPLES, ITALY 2. CNR,IST CIBERNETICA, I-80072 ARCO FELICE, NAPLES, ITALY (literal)

Titolo

Biosynthesis, release and degradation of the novel endogenous cannabimimetic metabolite 2-arachidonoylglycerol in mouse neuroblastoma cells (literal)

Abstract

The monoacylglycerol 2-arachidonoylglycerol (2-AG) has been recently suggested as a possible endogenous agonist at cannabinoid receptors both in brain and peripheral tissues. Here we report that a widely used model for neuronal cells, mouse N(18)TG(2) neuroblastoma cells, which contain the CB1 cannabinoid receptor, also biosynthesize, release and degrade 2-AG. Stimulation with ionomycin (1-5 mu M) of intact cells prelabelled with [H-3]arachidonic acid ([H-3]AA) led to the formation of high levels of a radioactive component with the same chromatographic behaviour as synthetic standards of 2-AG in TLC and HPLC analyses. The amounts of this metabolite were negligible in unstimulated cells, and greatly decreased in cells stimulated in the presence of the Ca2+-chelating agent EGTA. The purified component was further characterized as 2-AG by: (1) digestion with Rhizopus arrhizus lipase, which yielded radiolabelled AA; (2) gas chromatographic-MS analyses; and (3) TLC analyses on berate-impregnated plates. Approx. 20% of the 2-AG produced by stimulated cells was found to be released into the incubation medium when this contained 0.1% BSA. Subcellular fractions of N,,TG, cells were shown to contain enzymic activity or activities catalysing the hydrolysis of synthetic [H-3]2-AG to [H-3]AA. Cell homogenates were also found to convert synthetic [H-3]sn-1-acyl-2-arachidonoylglycerols (AcAGs) into [H-3]2-AG, suggesting that 2-AG might be derived from AcAG hydrolysis. When compared with ionomycin stimulation, treatment of cells with exogenous phospholipase C, but not with phospholipase D or A(2), led to a much higher formation of 2-AG and AcAGs. However, treatment of cells with phospholipase A(2) 10 min before ionomycin stimulation caused a 2.5-3-fold potentiation of 2-AG and AcAG levels with respect to ionomycin alone, whereas preincubation with the phospholipase C inhibitor neomycin sulphate did not inhibit the effect of ionomycin on 2-AG and AcAG levels. These results suggest that the Ca2+-induced formation of 2-AG proceeds through the intermediacy of AcAGs but not necessarily through phospholipase C activation. By showing for the first time the existence of molecular mechanisms for the inactivation and the Ca2+-dependent biosynthesis and release of 2-AG in neuronal cells, the present paper supports the hypothesis that this cannabimimetic monoacylglycerol might be a physiological neuromodular. (literal)

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