Dual effect of hydrogen peroxide on store-mediated calcium entry in human platelets.
Redox regulation is important for the modulation of cytosolic Ca(2+) concentration. Hence, we have investigated the effect of H(2)O(2) on store-mediated Ca(2+) entry (SMCE). In fura-2-loaded human platelets treatment with H(2)O(2) resulted in a concentration-dependent increase in Ca(2+) release from intracellular stores, while the effect on Ca(2+) entry was biphasic. In addition, 1mM H(2)O(2) reduced SMCE induced by agonists. The inhibitory effect of 1mM H(2)O(2) was prevented by inhibition of actin polymerization with cytochalasin D. Consistent with this, we found that 10microM H(2)O(2) and store depletion by treatment with thapsigargin plus ionomycin induced a similar temporal sequence of actin reorganization, while exposure to 1mM H(2)O(2) shifted the dynamics between polymerization and depolymerization in favor of the former. One millimolar H(2)O(2)-induced polymerization was reduced by treatment with methyl 2,5-dihydroxycinnamate and farnesylthioacetic acid, inhibitors of tyrosine kinases and Ras superfamily proteins, respectively. Finally, exposure to 1mM H(2)O(2) significantly increased store depletion-induced p60(src) activation. We conclude that H(2)O(2) exerted a biphasic effect on SMCE. The inhibitory role of high H(2)O(2) concentrations is mediated by an abnormal actin reorganization pattern involving both Ras- and tyrosine kinases-dependent pathways.
Redondo
Pedro C
PC
Department of Physiology, Faculty of Veterinary Sciences, University of Extremadura, 10071 Cáceres, Spain. jarosado@unex.es
Salido
Ginés M
GM
Pariente
José A
JA
Rosado
Juan A
JA
D000199:Actins / Q000378:metabolism; D001704:Biopolymers / Q000378:metabolism; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D015638:Cytochalasin D / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D015688:Oncogene Protein pp60(v-src) / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D018631:ras Proteins / Q000378:metabolism
D000199:Actins; D001704:Biopolymers; D019384:Nucleic Acid Synthesis Inhibitors; D015638:Cytochalasin D; D006861:Hydrogen Peroxide; D011505:Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src); D018631:ras Proteins; D002118:Calcium