Adenosine-5’-triphosphate (ATP) plays a pivotal role in various tissues as an extracellular transmitter. ATP released from nerve endings and/or damaged cells may elicit reactions in adjacent cells. To identify such reactions, we investigated the dynamics of the intracellular calcium ion concentrations ([Ca
2+]
i) in the rabbit corneal epithelium during ATP-stimulation. Intact epithelial sheets isolated from corneal tissue were loaded with Fura-2, and [Ca
2+]
i dynamics in each cell layer were analyzed using a digital imaging system (Argus 50/CA). Normal architecture was preserved, suggesting that functional integrity remained intact. Perfusion with HEPES-buffered Ringer’s solution containing ATP (10 μM) and uridine-5’-triphosphate (UTP; 10 μM) caused a biphasic [Ca
2+]
i increase in the superficial layer that manifested itself as a rapid initial spike followed by a long-lasting plateau phase. Adenosine-5’-diphate (10 μM) elevated the [Ca
2+]
i level, but induced only the initial spike, which was smaller than those induced by ATP and UTP. Adenosine (10 μM) did not elicit any [Ca
2+]
i changes in the epithelial cells. Suramin (10 μM; a P2 receptor antagonist) blocked the ATP-induced [Ca
2+]
i increase, whereas the P2X receptor agonists, α,β-methylene ATP (10 μM), 2-methylthio ATP (10 μM) and Benzoylbenzoyl ATP (10 μM), did not elicit any increases in [Ca
2+]
i. In the basal cell layer, ATP-induced [Ca
2+]
i dynamics were biphasic, while oscillatory fluctuations of [Ca
2+]
i were induced in the wing cells of the mid layer of the corneal epithelium by ATP stimulation. Ca
2+ oscillations were sometimes synchronized among adjacent wing cells, but these waves did not propagate to other cell layers. These results suggest that extracellular ATP elicits a [Ca
2+]
i increase mainly via P2Y receptors. In addition, synchronized Ca
2+ oscillation in the wing cell layer indicates that intracellular events may spread to neighboring cells within the layer.
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