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Am J Physiol Heart Circ Physiol (April 24, 2003). doi:10.1152/ajpheart.01155.2002
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Submitted on December 31, 2002
Accepted on April 22, 2003

Endogenously released ATP mediates shear-stress-induced Ca2+ influx into pulmonary artery endothelial cells

Kimiko Yamamoto1, Takaaki Sokabe1, Norihiko Ohura1, Hideki Nakatsuka1, Akira Kamiya2, and Joji Ando1*

1 Department of Biomedical Engineering, Graduate School of Medicine, University of Tokyo, Tokyo, Japan
2 Interdisciplinary Science Center, Nihon University, Tokyo, Japan

* To whom correspondence should be addressed. E-mail: joji{at}m.u-tokyo.ac.jp.

The mechanisms by which flow-imposed shear stress elevates intracellular Ca2+ in cultured endothelial cells (ECs) are not fully understood. Here we report finding that endogenously released ATP contributes to shear stress-induced Ca2+ responses. Application of flow of Hanks' balanced solution to human pulmonary artery ECs (HPAECs) elicited shear stress-dependent increase in Ca2+ concentrations. Chelation of extracellular Ca2+ with EGTA completely abolished the Ca2+ responses, whereas the phospholipase C inhibitor U-73122 or the Ca2+-ATPase inhibitor thapsigargin had no effect, indicating that the response was due to the influx of extracellular Ca2+. The Ca2+ influx was significantly suppressed by apyrase, which degrades ATP, or antisense oligonucleotide targeted to P2X4 receptors. A luciferase luminometric assay showed that shear stress induced dose-dependent release of ATP. When the ATP release was inhibited by the ATP synthase inhibitors angiostatin or oligomycin, the Ca2+ influx was markedly suppressed, but restored by removal of these inhibitors or addition of extracellular ATP. These results suggest that shear stress stimulates HPAECs to release ATP, which activates Ca2+ influx via P2X4 receptors.




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