{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,16]],"date-time":"2026-03-16T11:27:01Z","timestamp":1773660421251,"version":"3.50.1"},"reference-count":31,"publisher":"American Physiological Society","issue":"3","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["American Journal of Physiology-Cell Physiology"],"published-print":{"date-parts":[[2001,3,1]]},"abstract":"<jats:p> Although Na<jats:sup>+<\/jats:sup>-K<jats:sup>+<\/jats:sup>-2Cl<jats:sup>\u2212<\/jats:sup> cotransport has been demonstrated in cultured bovine corneal endothelial cells, its presence and role in the native tissue have been disputed. Using RT-PCR we have now identified a partial clone of the cotransporter protein in freshly dissected as well as in cultured corneal endothelial and epithelial cells. The deduced amino acid sequence of this protein segment is 99% identical to that of the bovine isoform (bNKCC1). [<jats:sup>3<\/jats:sup>H]bumetanide binding shows that the cotransporter sites are located in the basolateral membrane region at a density of 1.6 pmol\/mg of protein, close to that in lung epithelium. Immunocytochemistry confirms the basolateral location of the cotransporter. We calculate the turnover rate of the cotransporter to be 83 s<jats:sup>\u22121<\/jats:sup>. Transendothelial fluid transport, determined from deepithelialized rabbit corneal thickness measurements, is partially inhibited (30%) by bumetanide in a dose-dependent manner. Our results demonstrate that Na<jats:sup>+<\/jats:sup>-K<jats:sup>+<\/jats:sup>-2Cl<jats:sup>\u2212<\/jats:sup> cotransporters are present in the basolateral domain of freshly dissected bovine corneal endothelial cells and contribute to fluid transport across corneal endothelial preparations. <\/jats:p>","DOI":"10.1152\/ajpcell.2001.280.3.c491","type":"journal-article","created":{"date-parts":[[2017,12,24]],"date-time":"2017-12-24T20:24:39Z","timestamp":1514147079000},"page":"C491-C499","source":"Crossref","is-referenced-by-count":21,"title":["Corneal endothelial NKCC: molecular identification, location, and contribution to fluid transport"],"prefix":"10.1152","volume":"280","author":[{"given":"Kunyan","family":"Kuang","sequence":"first","affiliation":[{"name":"Departments of Ophthalmology,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yansui","family":"Li","sequence":"additional","affiliation":[{"name":"Departments of Ophthalmology,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Quan","family":"Wen","sequence":"additional","affiliation":[{"name":"Departments of Ophthalmology,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zheng","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, State University of New York-Optometry, New York, New York 10010"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Li","sequence":"additional","affiliation":[{"name":"Departments of Ophthalmology,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yingqing","family":"Yang","sequence":"additional","affiliation":[{"name":"Departments of Ophthalmology,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pavel","family":"Iserovich","sequence":"additional","affiliation":[{"name":"Departments of Ophthalmology,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter S.","family":"Reinach","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, State University of New York-Optometry, New York, New York 10010"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Janet","family":"Sparrow","sequence":"additional","affiliation":[{"name":"Anatomy and Cell Biology, Columbia University, New York 10032;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Friedrich P. 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