ATP-Induced Oscillations of Cytosolic Ca2+ Activity in Cultured Astrocytes from Rat Brain are Modulated by Medium Osmolarity Indicating a Control of [Ca2+]i Oscillations by Cell Volume

Oscillations of cytosolic Ca2+ activity ([Ca2+]i) induced by stimulation with ATP in rat astrocytes in primary cultures were analysed. Astrocytes, prepared from the brains of newborn rats, loaded with the fluorescent Ca2+ indicator fura-2/AM, were continuously stimulated with ATP (10 μM). ATP caused a large initial [Ca2+ peak, followed by regular [Ca2+]i oscillations (frequencies 1–5/min). Astrocytes were identified by glial fibrillary acidic protein staining of cells after [Ca2+]i recording. The oscillations were reversibly blocked by the P2 purinoceptor antagonist suramin (30 μM). Influx of extracellular Ca2+ and mobilization of Ca2+ from intracellular stores both contributed to the oscillations. The effects of hypertonic and hypotonic superfusion medium on ATP-induced [Ca2+]i oscillations were examined. Hypertonic medium (430 mOsm) reversibly suppressed the ATP-induced oscillations. Hypotonic medium (250 mOsm), in spite of having heterogeneous effects, most frequently induced a rise in [Ca2+]i, or reversibly increased the frequency of the oscillations. Thus, a change in cell volume might be closely connected with [Ca2+]i oscillations in astrocytes indicating that [Ca2+]i oscillations in glial cells play an important role in regulatory volume regulation in the brain.