Cerebral calcium accumulation and increases in the astroglial intermediate filament protein, glial fibrillary acidic protein (GFAP), have been used as markers of neurotoxic and ischemic brain damage. The present study was aimed at quantitatively investigating the regional and temporal relationship of those indices following a neurotoxic insult. For this purpose, regional changes in 45Ca uptake and GFAP levels, using ELISA, were evaluated in rat brains at both early (several hours) and late time points (up to 6 months) after a single systemic injection of kainic acid (12 mg/kg). After 4 h, limbic brain areas were already heavily labelled by 45Ca. In most investigated brain areas 45Ca accumulation peaked at day 4 (maximum 5 fold increase in amygdala) and returned to normal levels within 1 week (cerebellum, pons/medulla, occipital cortex), 2 weeks (striatum, frontal cortex), 2 or 4 months (limbic brain areas), or remained significantly elevated until 6 months (thalamus). In contrast, in all investigated brain areas, except cerebellum and pons/medulla, GFAP was increased from day 2, reaching maximum levels at day 28 in most limbic structures and remained significantly elevated at the same high level (15 fold increase) in amygdala, or somewhat lower levels in other affected regions (2-7 fold), but not in the thalamus. In all brain areas with 45Ca accumulation, GFAP was increased and the peak responses were highly correlated. Thus, both indices are useful quantitative biochemical markers of acute or subchronic neurotoxicity.