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Am J Physiol Heart Circ Physiol 285: H793-H803, 2003. First published April 24, 2003; doi:10.1152/ajpheart.01155.2002
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Endogenously released ATP mediates shear stress-induced Ca2+ influx into pulmonary artery endothelial cells

Kimiko Yamamoto,1 Takaaki Sokabe,1 Norihiko Ohura,1 Hideki Nakatsuka,1 Akira Kamiya,2 and Joji Ando1

1Department of Biomedical Engineering, Graduate School of Medicine, University of Tokyo; and 2Interdisciplinary Science Center, Nihon University, Tokyo 113-0033, Japan

Submitted 31 December 2002 ; accepted in final form 22 April 2003

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 increases 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, which thereby indicates 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 purinoceptors. 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 was 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.

hemodynamic force; mechanotransduction; ATP synthase; purinoceptors; and P2X4 receptor


Address for reprint requests and other correspondence: J. Ando, Dept. of Biomedical Engineering, Graduate School of Medicine, Univ. of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan (E-mail: joji{at}m.u-tokyo.ac.jp).




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