{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,13]],"date-time":"2025-09-13T16:30:42Z","timestamp":1757781042177},"reference-count":87,"publisher":"American Physiological Society","issue":"2","funder":[{"name":"Biomedical Sciences Ph.D. Program"},{"name":"Department of Pharmacology & Toxicology"},{"name":"Wright State Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["American Journal of Physiology-Cell Physiology"],"published-print":{"date-parts":[[2019,2,1]]},"abstract":"<jats:p> During aging, and development of atherosclerosis and cardiovascular disease (CVD), aortic vascular smooth muscle cells (VSMCs) transition from healthy contractile to diseased synthetic phenotypes. K-Cl cotransport (KCC) maintains cell volume and ion homeostasis in growth and differentiation, and hence is important for VSMC proliferation and migration. Therefore, KCC activity may play a role in the contractile-to-synthetic VSMC phenotypic transition. Early, medium, and late synthetic passage VSMCs were tested for specific cytoskeletal protein expression. KCC-mediated ouabain- and bumetanide-insensitive Rb<jats:sup>+<\/jats:sup> (a K<jats:sup>+<\/jats:sup> congener) influx was determined as Cl<jats:sup>\u2212<\/jats:sup>-dependent Rb<jats:sup>+<\/jats:sup> influx at different external Rb<jats:sup>+<\/jats:sup> and Cl<jats:sup>\u2212<\/jats:sup> ion concentrations, [Rb<jats:sup>+<\/jats:sup>]<jats:sub>o<\/jats:sub> and [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>o<\/jats:sub>. Expressions of the cytoskeletal proteins \u03b1-actin, vimentin, and desmin fell from early through late synthetic VSMCs. KCC kinetic parameters, such as maximum velocity ( V<jats:sub>m<\/jats:sub>), and apparent Cl<jats:sup>\u2212<\/jats:sup> and Rb<jats:sup>+<\/jats:sup> affinities ( K<jats:sub>m<\/jats:sub>), were calculated with Lineweaver-Burk, Hanes-Woolf, and Hill approximations. V<jats:sub>m<\/jats:sub> values of both Rb<jats:sup>+<\/jats:sup>- and Cl<jats:sup>\u2212<\/jats:sup>-dependent influxes were of equal magnitude, commensurate with a KCC stoichiometry of unity, and rose threefold from early to late synthetic VSMCs. Hill coefficients for Rb<jats:sup>+<\/jats:sup> and Cl<jats:sup>\u2212<\/jats:sup> correlated with cell passage number, suggesting increased KCC ligand cooperativity. However, K<jats:sub>m<\/jats:sub> values for [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>o<\/jats:sub> were strikingly bimodal with 60\u201380 mM in early, ~20\u201330 mM in medium, and 60 mM in late passage cells. In contrast, K<jats:sub>m<\/jats:sub> values for [Rb<jats:sup>+<\/jats:sup>]<jats:sub>o<\/jats:sub> remained steady at ~17 mM. Since total KCC isoform expression was similar with cell passage, structure\/function changes of the KCC signalosome may accompany the transition of aortic VSMCs from a healthy to a diseased phenotype. <\/jats:p>","DOI":"10.1152\/ajpcell.00002.2017","type":"journal-article","created":{"date-parts":[[2019,1,16]],"date-time":"2019-01-16T20:05:50Z","timestamp":1547669150000},"page":"C274-C284","source":"Crossref","is-referenced-by-count":8,"title":["Kinetic studies of K-Cl cotransport in cultured rat vascular smooth muscle cells"],"prefix":"10.1152","volume":"316","author":[{"given":"Peter K","family":"Lauf","sequence":"first","affiliation":[{"name":"The Cell Biophysics Group, Wright State University, Dayton, Ohio"},{"name":"Department of Pharmacology and Toxicology, Wright State University, Dayton, Ohio"},{"name":"Department of Pathology, Wright State University, Dayton, Ohio"},{"name":"Boonshoft School of Medicine, Wright State University, Dayton, 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