The effects of glucose deprivation, hypoxia and glucose-free hypoxia conditions on phosphoinositide (PI) hydrolysis were studied in cortical slices from 8-day-old rats. Only glucose-free hypoxia induced a significant increase of inositol phosphate formation. The inositol phosphate formation induced by noradrenaline, carbachol and several excitatory amino acid receptor agonists, but not the Ca2+ ionophore A23187-induced stimulation, was blocked by glucose-free hypoxia and differentially reduced by glucose and oxygen deprivation depending on the neurotransmitter receptor agonist. The stimulatory effect of glucose-free hypoxia was not reduced by the muscarinic receptor antagonist atropine or by the inhibitors of the excitatory amino acid-stimulated PI hydrolysis DL-2-amino-3-phosphono-propionic acid and L-aspartate-beta-hydroxamate, and neither by the voltage-sensitive Na+ channel tetrodotoxin. The effect of glucose-free hypoxia was partially dependent on extracellular Ca2+ and it was blocked by verapamil and amiloride, but not by nifedipine, Co2+ and neomycin. These results suggest that Ca2+ influx through the Na(+)-Ca2+ exchanger underlies the PI hydrolysis stimulation induced by combined glucose and oxygen deprivation in neonatal cerebral cortical slices.