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Everardo Avelino

Connexin hemichannels-mediated Ca 2+ entry results in nitric oxide production in in situ injured endothelial cells. Everardo Avelino. Secondo anno Ciclo XXII. Introduction. Injury provokes a Ca 2+ wave characterized by two phases, peak and plateau. PEAK

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Everardo Avelino

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  1. Connexin hemichannels-mediated Ca2+ entry results in nitric oxide production in in situ injured endothelial cells Everardo Avelino Secondo anno Ciclo XXII

  2. Introduction Injury provokes a Ca2+ wave characterized by two phases, peak and plateau PEAK ~ 60% > intracellular stores depletion (P2Y1,2,12,13) ~ 40 > Ca2+ influx (P2X and Gap junctions) PLATEAU Ca2+ influx (Gap junctions)

  3. Introduction • Endothelium functions: • Angiogenesis and vasculogenesis • Preserve a surface unreactive toward circulating cells • Mantain thromboresistence • Inhibit vascular tone • Inhibit cells growth in outer layer of the vesell wall.

  4. Nitric oxide measurement DAF FM DA Fluorescent dye Indicator of NO production Emission: 510nm Excitation: 490nm

  5. Injury augments nitric oxide production in rat aortic endothelium

  6. Injury augments nitric oxide production in rat aortic endothelium

  7. Injury induced NOP* depend on extracellular Ca2+ * Nitric oxide production

  8. Capacitative Ca2+ entry does not sustain injury-induced NOP

  9. Putative blockers of CxHcs* reduce NO synthesis induced by Ca2+ entry in injured endothelium *Connexin hemichannels

  10. Putative blockers of CxHcs* reduce NO synthesis induced by Ca2+ entry in injured endothelium *Connexin hemichannels

  11. Connexin mimetic peptides reduce both injury-induced Ca2+ elevation and NO production in ra aortic EC’s Pre-incubation

  12. Connexin mimetic peptides reduce both injury-induced Ca2+ elevation and NO production in ra aortic EC’s Pre-incubation

  13. Connexin mimetic peptides reduce both injury-induced Ca2+ elevation and NO production in ra aortic EC’s Accute effect

  14. Connexin mimetic peptides reduce both injury-induced Ca2+ elevation and NO production in ra aortic EC’s

  15. Summary

  16. Summary ? Ca2+ ATPase ATP ATP Activates NCCE Inhibit CCE TRPV4 eNOS PLC PG ATP CaM IP3 Ca2+ Ca2+ Gap Ca2+ IP3R Intracellular Ca2+ stores Gap Nucleus Ca2+

  17. Preeliminar results

  18. Preeliminar results

  19. Preeliminar results

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