The effect of acute alcohol (33 mM ethanol) on calcium signaling evoked by glutamate receptor activation was studied in cultured cerebellar Purkinje and granule neurons at different stages of development. Calcium signals were measured by microscopic imaging using the calcium sensitive dye fura-2. At an early stage in development (10 days in vitro), acute alcohol enhanced the calcium signals evoked in Purkinje neurons by exogenous application of quisqualate, an agonist at ionotropic and metabotropic glutamate receptors. In contrast, in mature cultured Purkinje neurons (21-24 days in vitro) the calcium signals produced by quisqualate were reduced by alcohol. At an intermediate stage of development (14 days in vitro) reflecting the main period of morphological and physiological maturation, alcohol had no significant effect on the response to quisqualate. Alcohol's actions were significantly altered by manipulation of the intracellular stores with caffeine, implicating intracellular stores in alcohol's actions. Calcium signals produced by quisqualate in the cultured granule neurons were also altered by acute alcohol, in a manner similar to that observed in the Purkinje neurons. These data demonstrate that calcium signaling pathways are a site of alcohol action in developing CNS neurons and that the cellular consequences of alcohol exposure can change with development. Such actions of alcohol could have significant effects on the immature nervous system, where the precise timing of appropriate signaling levels are important aspects of the maturation process.