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Papers In Press, published online ahead of print December 21, 2005
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Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Osaka 567-0047
Corresponding Author: akihito{at}sanken.osaka-u.ac.jp
Sphingosine 1-phosphate (S1P) is accumulated in platelets and released on stimulation by thrombin or Ca2+. Thrombin-stimulated S1P release was inhibited by staurosporin, whereas Ca2+-stimulated release was not. When the platelet plasma membrane was permeabilized with streptolysin O (SLO), S1P leaked out with cytosol markers, while granular markers remained in the platelets. The SLO-induced S1P leakage required bovine serum albumin (BSA), probably for solubilization of S1P in the medium. These results indicate that S1P is localized in the inner leaflet of the plasma membrane and that its release is a carrier-mediated process. We also used alpha-toxin (ATX) that makes smaller pores in the plasma membrane than SLO, and depletes cytosolic ATP without BSA-dependent S1P leakage. The addition of ATP drove S1P release from ATX-platelets. The ATP-driven S1P release from ATX-platelets was greatly enhanced by thrombin. An ATP binding cassette (ABC) transporter inhibitor, glyburide, prevents ATP- and thrombin-induced S1P release from platelet. Ca2+ also stimulated S1P release from ATX-platelets without ATP, while the Ca2+-induced release was not inhibited by glyburide. Our results indicate that two independent S1P release systems might exist in the platelet plasma membrane, an ATP-dependent system stimulated by thrombin and an ATP-independent one stimulated by Ca2+.
Revised on December 16, 2005
Accepted on December 21, 2005
Sphingosine 1-phosphate is released from the cytosol of rat platelets in a carrier-mediated manner
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