{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T13:04:02Z","timestamp":1791378242946,"version":"4.3.2"},"reference-count":101,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2018,4,15]],"date-time":"2018-04-15T00:00:00Z","timestamp":1523750400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["R01 DK\u2010091336"],"award-info":[{"award-number":["R01 DK\u2010091336"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["physoc.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Physiology"],"published-print":{"date-parts":[[2018,4,15]]},"abstract":"<jats:sec><jats:title>Key points<\/jats:title><jats:p><jats:list list-type=\"bullet\"><jats:list-item><jats:p>Contraction of urethral smooth muscle cells (USMCs) contributes to urinary continence. Ca<jats:sup>2+<\/jats:sup>signalling in USMCs was investigated in intact urethral muscles using a genetically encoded Ca<jats:sup>2+<\/jats:sup>sensor, GCaMP3, expressed selectively in USMCs.<\/jats:p><\/jats:list-item><jats:list-item><jats:p>USMCs were spontaneously active<jats:italic>in situ<\/jats:italic>, firing intracellular Ca<jats:sup>2+<\/jats:sup>waves that were asynchronous at different sites within cells and between adjacent cells.<\/jats:p><\/jats:list-item><jats:list-item><jats:p>Spontaneous Ca<jats:sup>2+<\/jats:sup>waves in USMCs were myogenic but enhanced by adrenergic or purinergic agonists and decreased by nitric oxide.<\/jats:p><\/jats:list-item><jats:list-item><jats:p>Ca<jats:sup>2+<\/jats:sup>waves arose from inositol trisphosphate type 1 receptors and ryanodine receptors, and Ca<jats:sup>2+<\/jats:sup>influx by store\u2010operated calcium entry was required to maintain Ca<jats:sup>2+<\/jats:sup>release events.<\/jats:p><\/jats:list-item><jats:list-item><jats:p>Ca<jats:sup>2+<\/jats:sup>release and development of Ca<jats:sup>2+<\/jats:sup>waves appear to be the primary source of Ca<jats:sup>2+<\/jats:sup>for excitation\u2013contraction coupling in the mouse urethra, and no evidence was found that voltage\u2010dependent Ca<jats:sup>2+<\/jats:sup>entry via L\u2010type or T\u2010type channels was required for responses to \u03b1 adrenergic responses.<\/jats:p><\/jats:list-item><\/jats:list><\/jats:p><\/jats:sec><jats:sec><jats:title>Abstract<\/jats:title><jats:p>Urethral smooth muscle cells (USMCs) generate myogenic tone and contribute to urinary continence. Currently, little is known about Ca<jats:sup>2+<\/jats:sup>signalling in USMCs<jats:italic>in situ<\/jats:italic>, and therefore little is known about the source(s) of Ca<jats:sup>2+<\/jats:sup>required for excitation\u2013contraction coupling. We characterized Ca<jats:sup>2+<\/jats:sup>signalling in USMCs within intact urethral muscles using a genetically encoded Ca<jats:sup>2+<\/jats:sup>sensor, GCaMP3, expressed selectively in USMCs. USMCs fired spontaneous intracellular Ca<jats:sup>2+<\/jats:sup>waves that did not propagate cell\u2010to\u2010cell across muscle bundles. Ca<jats:sup>2+<\/jats:sup>waves increased dramatically in response to the \u03b11 adrenoceptor agonist phenylephrine (10\u00a0\u03bc<jats:sc>m<\/jats:sc>) and to ATP (10\u00a0\u03bc<jats:sc>m<\/jats:sc>). Ca<jats:sup>2+<\/jats:sup>waves were inhibited by the nitric oxide donor DEA NONOate (10\u00a0\u03bc<jats:sc>m<\/jats:sc>). Ca<jats:sup>2+<\/jats:sup>influx and release from sarcoplasmic reticulum stores contributed to Ca<jats:sup>2+<\/jats:sup>waves, as Ca<jats:sup>2+<\/jats:sup>free bathing solution and blocking the sarcoplasmic Ca<jats:sup>2+<\/jats:sup>\u2010ATPase abolished activity. Intracellular Ca<jats:sup>2+<\/jats:sup>release involved cooperation between ryanadine receptors and inositol trisphosphate receptors, as tetracaine and ryanodine (100\u00a0\u03bc<jats:sc>m<\/jats:sc>) and xestospongin C (1\u00a0\u03bc<jats:sc>m<\/jats:sc>) reduced Ca<jats:sup>2+<\/jats:sup>waves. Ca<jats:sup>2+<\/jats:sup>waves were insensitive to L\u2010type Ca<jats:sup>2+<\/jats:sup>channel modulators nifedipine (1\u00a0\u03bc<jats:sc>m<\/jats:sc>), nicardipine (1\u00a0\u03bc<jats:sc>m<\/jats:sc>), isradipine (1\u00a0\u03bc<jats:sc>m<\/jats:sc>) and FPL 64176 (1\u00a0\u03bc<jats:sc>m<\/jats:sc>), and were unaffected by the T\u2010type Ca<jats:sup>2+<\/jats:sup>channel antagonists NNC\u2010550396 (1\u00a0\u03bc<jats:sc>m<\/jats:sc>) and TTA\u2010A2 (1\u00a0\u03bc<jats:sc>m<\/jats:sc>). Ca<jats:sup>2+<\/jats:sup>waves were reduced by the store operated Ca<jats:sup>2+<\/jats:sup>entry blocker SKF 96365 (10\u00a0\u03bc<jats:sc>m<\/jats:sc>) and by an Orai antagonist, GSK\u20107975A (1\u00a0\u03bc<jats:sc>m<\/jats:sc>). The latter also reduced urethral contractions induced by phenylephrine, suggesting that Orai can function effectively as a receptor\u2010operated channel. In conclusion, Ca<jats:sup>2+<\/jats:sup>waves in mouse USMCs are a source of Ca<jats:sup>2+<\/jats:sup>for excitation\u2013contraction coupling in urethral muscles.<\/jats:p><\/jats:sec>","DOI":"10.1113\/jp275719","type":"journal-article","created":{"date-parts":[[2018,1,31]],"date-time":"2018-01-31T05:04:49Z","timestamp":1517375089000},"page":"1433-1466","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":37,"title":["Ca<sup>2+<\/sup>signalling in mouse urethral smooth muscle<i>in situ<\/i>: role of Ca<sup>2+<\/sup>stores and Ca<sup>2+<\/sup>influx mechanisms"],"prefix":"10.1113","volume":"596","author":[{"given":"Bernard T.","family":"Drumm","sequence":"first","affiliation":[{"name":"Department of Physiology and Cell Biology University of Nevada, Reno School of Medicine Reno NV 89557 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin E.","family":"Rembetski","sequence":"additional","affiliation":[{"name":"Department of Physiology and Cell Biology University of Nevada, Reno School of Medicine Reno NV 89557 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2871-3825","authenticated-orcid":false,"given":"Caroline A.","family":"Cobine","sequence":"additional","affiliation":[{"name":"Department of Physiology and Cell Biology University of Nevada, Reno School of Medicine Reno NV 89557 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Salah A.","family":"Baker","sequence":"additional","affiliation":[{"name":"Department of Physiology and Cell Biology University of Nevada, Reno School of Medicine Reno NV 89557 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9443-5491","authenticated-orcid":false,"given":"Gerard P.","family":"Sergeant","sequence":"additional","affiliation":[{"name":"Smooth Muscle Research Centre Dundalk Institute of Technology Co. 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