{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T07:04:19Z","timestamp":1777964659532,"version":"3.51.4"},"reference-count":56,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2004,5,24]],"date-time":"2004-05-24T00:00:00Z","timestamp":1085356800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The Journal of Physiology"],"published-print":{"date-parts":[[2004,6]]},"abstract":"<jats:p>ClC\u20103, a member of the large superfamily of ClC voltage\u2010dependent Cl<jats:sup>\u2013<\/jats:sup> channels, has been proposed as a molecular candidate responsible for volume\u2010sensitive osmolyte and anion channels (VSOACs) in some cells, including heart and vascular smooth muscle. However, the reported presence of native VSOACs in at least two cell types from transgenic ClC\u20103 disrupted (<jats:italic>Clcn3<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup>) mice casts considerable doubt on this proposed role for ClC\u20103. We compared several properties of native VSOACs and examined mRNA transcripts and membrane protein expression profiles in cardiac and pulmonary arterial smooth muscle cells from <jats:italic>Clcn3<\/jats:italic><jats:sup>+\/+<\/jats:sup> and <jats:italic>Clcn3<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup> mice to: (1) test the hypothesis that native VSOACs are unaltered in cells from <jats:italic>Clcn3<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup> mice, and (2) test the possibility that targeted inactivation of the Clcn3 gene using a conventional murine global knock\u2010out approach may result in compensatory changes in expression of other membrane proteins. Our experiments demonstrate that VSOAC currents in myocytes from <jats:italic>Clcn3<\/jats:italic><jats:sup>+\/+<\/jats:sup> and <jats:italic>Clcn3<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup> mice are remarkably similar in terms of activation and inactivation kinetics, steady\u2010state current densities, rectification, anion selectivity (I<jats:sup>\u2212<\/jats:sup> &gt; Cl<jats:sup>\u2212<\/jats:sup>\u226b Asp<jats:sup>\u2212<\/jats:sup>) and sensitivity to block by glibenclamide, niflumic acid, DIDS and extracellular ATP. However, additional experiments revealed several significant differences in other fundamental properties of native VSOACs recorded from atrial and smooth muscle cells from <jats:italic>Clcn3<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup> mice, including: differences in regulation by endogenous protein kinase C, differential sensitivity to block by anti\u2010ClC\u20103 antibodies, and differential sensitivities to [ATP]<jats:sub>i<\/jats:sub> and free [Mg<jats:sup>2+<\/jats:sup>]<jats:sub>i<\/jats:sub>. These results suggest that in response to <jats:italic>Clcn3<\/jats:italic> gene deletion, there may be compensatory changes in expression of other proteins that alter VSOAC channel subunit composition or associated regulatory subunits that give rise to VSOACs with different properties. Consistent with this hypothesis, in atria from <jats:italic>Clcn3<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup> mice compared to <jats:italic>Clcn3<\/jats:italic><jats:sup>+\/+<\/jats:sup> mice, quantitative analysis of ClC mRNA expression levels revealed significant increases in transcripts for ClC\u20101, ClC\u20102, and ClC\u20103, and protein expression profiles obtained using two\u2010dimensional polyacrylamide gel electrophoresis revealed complex changes in at least 35 different unidentified membrane proteins in cells from <jats:italic>Clcn3<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup> mice. These findings emphasize that caution needs to be exercised in simple attempts to interpret the phenotypic consequences of conventional global <jats:italic>Clcn3<\/jats:italic> gene inactivation.<\/jats:p>","DOI":"10.1113\/jphysiol.2003.059261","type":"journal-article","created":{"date-parts":[[2004,3,16]],"date-time":"2004-03-16T01:44:34Z","timestamp":1079401474000},"page":"439-456","source":"Crossref","is-referenced-by-count":84,"title":["Altered properties of volume\u2010sensitive osmolyte and anion channels (VSOACs) and membrane protein expression in cardiac and smooth muscle myocytes from <i>Clcn3<\/i><sup>\u2010\/\u2010<\/sup> mice"],"prefix":"10.1113","volume":"557","author":[{"given":"Shintaro","family":"Yamamoto\u2010Mizuma","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ge\u2010Xin","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luis 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