{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T18:08:31Z","timestamp":1778695711149,"version":"3.51.4"},"reference-count":52,"publisher":"American Physiological Society","issue":"6","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["American Journal of Physiology-Cell Physiology"],"published-print":{"date-parts":[[2008,6]]},"abstract":"<jats:p> In this study, we have systematically evaluated the signaling mechanisms underlying stimulated nitric oxide (NO) synthesis by estrogen (E<jats:sub>2<\/jats:sub>) and other vasoactive agents at the level of a single endothelium-derived cell. To do so, we have characterized and contrasted rapid E<jats:sub>2<\/jats:sub>-evoked NO synthesis with that of ATP using single-cell microfluorimetry and patch-clamp recordings to monitor stimulated changes in cellular NO synthesis (via 4-amino-5-methylamino-2\u2032,7\u2032-difluorofluorescein), Ca<jats:sup>2+<\/jats:sup> transients (via Fluo-3), and membrane hyperpolarization in cultured human EA.hy926 cells. E<jats:sub>2<\/jats:sub>-evoked NO synthesis in single cells (EC<jats:sub>50<\/jats:sub> \u223c0.3 nM) was blocked by the E<jats:sub>2<\/jats:sub> receptor antagonist ICI 182,780 and the NO synthase inhibitor N<jats:sup>\u03c9<\/jats:sup>-nitro-l-arginine methyl ester. Although both E<jats:sub>2<\/jats:sub> and ATP stimulated comparable Ca<jats:sup>2+<\/jats:sup> transients, E<jats:sub>2<\/jats:sub>-induced NO synthesis was insensitive to intracellular BAPTA-AM or removal of external Ca<jats:sup>2+<\/jats:sup>. In contrast, ATP-evoked NO production was abolished by either one of these treatments. ATP-evoked hyperpolarizations (\u223c20 mV) and NO production were both inhibited by the respective small-conductance and intermediate-conductance calcium- activated K<jats:sup>+<\/jats:sup> channel blockers apamin and charybdotoxin. E<jats:sub>2<\/jats:sub> minimally affected membrane potential, and stimulated NO synthesis was insensitive to calcium-activated K<jats:sup>+<\/jats:sup> channel blockers. Exposure to either the phosphatidylinositol 3-kinase inhibitor LY-294001 or the MAP kinase inhibitor PD-98059 abolished the NO response to E<jats:sub>2<\/jats:sub>, but not that to ATP. Finally, the NO response evoked by a combined stimulus of E<jats:sub>2<\/jats:sub> plus ATP was similar to that of ATP alone. In conclusion, our data directly demonstrate that an individual human EA.hy926 cell contains at least two distinct mechanisms for stimulated NO synthesis that depend on either calcium or protein kinase signaling events. <\/jats:p>","DOI":"10.1152\/ajpcell.00561.2007","type":"journal-article","created":{"date-parts":[[2008,3,27]],"date-time":"2008-03-27T01:00:59Z","timestamp":1206579659000},"page":"C1531-C1541","source":"Crossref","is-referenced-by-count":11,"title":["Estrogen and the Ca<sup>2+<\/sup>-mobilizing agonist ATP evoke acute NO synthesis via distinct pathways in an individual human vascular endothelium-derived cell"],"prefix":"10.1152","volume":"294","author":[{"given":"Jian-Zhong","family":"Sheng","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Furqan","family":"Arshad","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Janice E.","family":"Braun","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew P.","family":"Braun","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"24","reference":[{"key":"R1","doi-asserted-by":"crossref","unstructured":"Alessi DR, Cuenda A, Cohen P, Dudley DT, Saltiel AR. 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