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The underlying physiological mechanisms driving this vasorelaxation remain unclear. Studies were designed to test the hypothesis that multiple signaling pathways including nitric oxide (NO) and endothelium\u2010derived hyperpolarizing factor (EDHF) in endothelial cells as well as Ca<jats:sup>2+<\/jats:sup> antagonization and TRPM7 channels in vascular smooth muscle cells mediate Mg<jats:sup>2+<\/jats:sup>\u2010dependent vessel relaxation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>To uncover these mechanisms, force development was measured ex\u00a0vivo in aorta rings from mice using isometric wire myography. Concentration responses to Mg<jats:sup>2+<\/jats:sup> were studied in intact and endothelium\u2010denuded aortas. Key findings were confirmed in second\u2010order mesenteric resistance arteries perfused ex\u00a0vivo using pressure myography. Effects of Mg<jats:sup>2+<\/jats:sup> on NO formation were measured in Chinese Hamster Ovary (CHO) cells, isolated mesenteric vessels, and mouse urine.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Mg<jats:sup>2+<\/jats:sup> caused a significant concentration\u2010dependent relaxation of aorta rings. This relaxation was attenuated significantly in endothelium\u2010denuded aortas. The endothelium\u2010dependent portion was inhibited by NO and cGMP blockade but not by cyclooxygenase inhibition. Mg<jats:sup>2+<\/jats:sup> stimulated local NO formation in CHO cells and isolated mesenteric vessels without changing urinary NOx levels. High extracellular Mg<jats:sup>2+<\/jats:sup> augmented acetylcholine\u2010induced relaxation. SK<jats:sub>Ca<\/jats:sub> and IK<jats:sub>Ca<\/jats:sub> channel blockers apamin and TRAM34 inhibited Mg<jats:sup>2+<\/jats:sup>\u2010dependent relaxation. The endothelium\u2010independent relaxation in aorta rings was inhibited by high extracellular Ca<jats:sup>2+<\/jats:sup>. Combined blockade of NO, SK<jats:sub>Ca<\/jats:sub>, and IK<jats:sub>Ca<\/jats:sub> channels significantly reduced Mg<jats:sup>2+<\/jats:sup>\u2010dependent dilatation in mesenteric resistance vessels.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>In mouse conductance and resistance arteries Mg<jats:sup>2+<\/jats:sup>\u2010induced relaxation is contributed by endothelial NO formation, EDHF pathways, antagonism of Ca<jats:sup>2+<\/jats:sup> in smooth muscle cells, and additional unidentified mechanisms.<\/jats:p><\/jats:sec>","DOI":"10.1111\/apha.14096","type":"journal-article","created":{"date-parts":[[2024,1,23]],"date-time":"2024-01-23T10:05:21Z","timestamp":1706004321000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Nitric oxide, endothelium\u2010derived hyperpolarizing factor, and smooth muscle\u2010dependent mechanisms contribute to magnesium\u2010dependent vascular relaxation 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