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Am J Physiol Cell Physiol 274: C1456-C1465, 1998;
0363-6143/98 $5.00
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Vol. 274, Issue 6, C1456-C1465, June 1998

IP3 receptor blockade fails to prevent intracellular Ca2+ release by ET-1 and alpha -thrombin

Robert S. Mathias1, Katsuhiko Mikoshiba2, Takayuki Michikawa2, Atsushi Miyawaki2, and Harlan E. Ives3,4

Departments of 1 Pediatrics and 3 Medicine and 4 Cardiovascular Research Institute, University of California, San Francisco, California 94143; and 2 Department of Molecular Neurobiology, Institute of Medical Science, University of Tokyo, Tokyo, Japan

The effect of inositol 1,4,5-trisphosphate (IP3) receptor blockade on platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), endothelin-1 (ET-1), or alpha -thrombin receptor-mediated intracellular Ca2+ (Ca2+i) release was examined using fura 2 microspectrofluorometry in single Chinese hamster ovary cells and myoblasts. Blockade of the IP3 receptor was achieved by microinjection of heparin or monoclonal antibody (MAb) 18A10 into the IP3 type 1 receptor. Heparin completely inhibited Ca2+i release after flash photolysis with caged IP3 and after exposure to PDGF and FGF. In contrast, heparin failed to block Ca2+i release after alpha -thrombin and ET-1. After application of ligand, IP3 levels were five- to sevenfold higher for alpha -thrombin than for ET-1 or PDGF. IP3 levels after PDGF and ET-1 were comparable. Similar to heparin, MAb 18A10 blocked Ca2+i release after PDGF but failed to block Ca2+i release after ET-1 or alpha -thrombin. These data suggest that the mechanisms of Ca2+i release by tyrosine kinase and certain 7-transmembrane receptors may differ. Although both receptor types use the IP3-signaling system, the ET-1 and alpha -thrombin receptors may have a second, alternative mechanism for activating Ca2+i release.

platelet-derived growth factor; endothelin-1; intracellular microinjection of heparin and monoclonal antibody 18A10; Chinese hamster ovary cells; inositol 1,4,5-trisphosphate


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