We studied the effect of external Ca2+ on the incorporation of [32P]phosphate into the astrocytic marker protein, glial fibrillary acidic protein (GFAP), in hippocampal slices from rats in the postnatal age range 12-16 days to +60 days (P12-P16 to +P60). At age P12-P16 the presence of Ca2+ in the incubation medium inhibited the incorporation of 32P into GFAP; this inhibition declined to near zero by P21 and subsequently 32P-incorporation became progressively more dependent on Ca2+ until by P60 no GFAP phosphorylation was observed in the absence of Ca2+. With tissue from immature rats inhibition of 32P-incorporation into GFAP started at a medium concentration of 7.5 microM Ca2+, reached 50% at 100 microM and then remained constant up to 1 mM; with adults maximal phosphorylation required 1 mM Ca2+ in the medium. The inorganic Ca(2+)-channel blockers, Co2+ and Ni2+, and a high concentration of the L-type blocker, nifedipine, reversed the effects of external Ca2+ on GFAP phosphorylation. The results suggest a late developmental change in the compartmental disposition of Ca2+ in astrocytes.