The effect of 5-hydroxytryptamine (5HT) on cytoplasmic Ca2+ concentration was examined at the single cell level in astroglial enriched primary cultures from newborn rat cerebral cortex. Type 1 astroglial cells were identified and the Ca2+ indicator dye fura 2/AM was used in a microspectrofluorimetric system. Pharmacological studies indicated that the Ca2+ responses were mediated by 5HT2 receptors. Four different patterns of 5HT evoked cytosolic Ca2+ responses were identified including two different types of spike patterns and two types of Ca2+ oscillations (low amplitude and base-line spiking behaviour). In addition, cells with spontaneous Ca2+ oscillations were of two types, those responding to 5HT and those not responding to 5HT. The different responses were identified already on day 7 in culture and were followed up to day 21 with a concomitant increase in the number of responding cells, although the response patterns did not differ during culture. The triggering 5HT concentration was 1 microM. The results suggest that subpopulations of astrocytes exist with respect to 5HT2-evoked cytosolic Ca2+ mobilization.