Endothelin-evoked Release of Arachidonic Acid from Mouse Astrocytes in Primary Culture.
In striatal astrocytes, receptors for the vasoactive peptide endothelin (ET) are associated with several intracellular signalling pathways: ET-1 increases the breakdown of phosphoinositides, induces a sustained influx of Ca2+ and inhibits the isoproterenol-induced formation of cAMP (Marin et al., J. Neurochem., 56, 1270 - 1275, 1991). In the present study, it will be shown that ET-1 and ET-3 markedly stimulate the release of arachidonic acid (AA) from cultured astrocytes from the mouse striatum (EC50=3 and 7 nM for ET-1 and ET-3, respectively), mesencephalon and cerebral cortex. The ET-1-evoked release of AA probably resulted from the activation of a phospholipase A2, since it required extracellular Ca2+ and was prevented by mepacrine but not by RHC 80267, an inhibitor of diacylglycerol lipase. The ET-1-induced release of AA was shown to be partially mediated by a guanine nucleotide-binding protein sensitive to pertussis toxin but not to cholera toxin. A cAMP-dependent process is not involved since the ET-1-evoked release of AA was not affected when cells were incubated with either isoproterenol or 8-bromo-cAMP. The ET-1-evoked release of AA could be mimicked by the co-application of a calcium ionophore and a protein kinase C activator. However, staurosporine, a potent inhibitor of protein kinase C, which blocked the release of AA induced by the combined application of ionomycin and phorbol 12-myristate 12-acetate (PMA), was without effect on the ET-1-evoked response, indicating that protein kinase C is not directly involved in the ET-1-induced release of AA. Furthermore, the responses induced by ET-1 and by PMA were found to be additive. These results suggest that (1) ET-1 receptors are coupled to the release of AA by a mechanism independent of both protein kinase C activation and the adenylate cyclase pathway, possibly via the activation of phospholipase A2, (2) different mechanisms (or different phospholipase A2 subtypes) are involved in the control of AA release in astrocytes.
Tencé
Martine
M
Laboratorie de Neuropharmacologie, INSERM U114, Collège de France, 11, Place Marcelin Berthelot, 75231 Paris Cedex 05, France.
Cordier
Jocelyne
J
Glowinski
Jacques
J
Prémont
Joël
J