{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,9]],"date-time":"2026-06-09T22:06:02Z","timestamp":1781042762811,"version":"3.54.1"},"reference-count":43,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2015,3,12]],"date-time":"2015-03-12T00:00:00Z","timestamp":1426118400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biology of the Cell"],"published-print":{"date-parts":[[2015,4]]},"abstract":"<jats:sec><jats:title>Background information<\/jats:title><jats:p>The gain\u2010of\u2010function A843E mutation of the calcium sensing receptor (CaR) causes Bartter syndrome type 5. Patients carrying this CaR variant show a remarkably reduced renal NaCl reabsorption in the thick ascending limb (TAL) of Henle's loop resulting in renal loss of NaCl in the absence of mutations in renal Na<jats:sup>+<\/jats:sup> and Cl<jats:sup>\u2212<\/jats:sup> ion transporters. The molecular mechanisms underlying this clinical phenotype are incompletely understood. We investigated, in human embryonic kidney 293 (HEK 293) cells and porcine kidney epithelial (LLC\u2010PK1) cells, the functional cross\u2010talk of CaR\u2010A843E with the Na<jats:sup>+<\/jats:sup>:K<jats:sup>+<\/jats:sup>:2Cl<jats:sup>\u2013<\/jats:sup> co\u2010transporter, NKCC2, which provides NaCl reabsorption in the TAL.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The expression of the CaR mutant did not alter the apical localisation of NKCC2 in LLC\u2010PK1 cells. However, the steady\u2010state NKCC2 phosphorylation and activity were decreased in cells transfected with CaR\u2010A843E compared with the control wild\u2010type CaR (CaR WT)\u2010transfected cells. Of note, low\u2010Cl<jats:sup>\u2212<\/jats:sup>\u2010dependent NKCC2 activation was also strongly inhibited upon the expression of CaR\u2010A843E mutant. The use of either P450 \u03c9\u2010hydroxylase (CYP4)\u2010 or phospholipase A2 (PLA2)\u2010blockers suggests that this effect is likely mediated by arachidonic acid (AA) metabolites.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The data suggested that the activated CaR affects intracellular pathways modulating NKCC2 activity rather than NKCC2 intracellular trafficking in renal cells, and throw further light on the pathological role played by active CaR mutants in Bartter syndrome type 5.<\/jats:p><\/jats:sec>","DOI":"10.1111\/boc.201400069","type":"journal-article","created":{"date-parts":[[2015,1,28]],"date-time":"2015-01-28T13:27:56Z","timestamp":1422451676000},"page":"98-110","source":"Crossref","is-referenced-by-count":19,"title":["NKCC2 activity is inhibited by the Bartter's syndrome type 5 gain\u2010of\u2010function CaR\u2010A843E mutant in renal cells"],"prefix":"10.1111","volume":"107","author":[{"given":"Monica","family":"Carmosino","sequence":"first","affiliation":[{"name":"Department of Biosciences, Biotechnologies, and Biopharmaceutics University of Bari  70125 Bari Italy"},{"name":"Department of Sciences University of Basilicata  85100 Potenza Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Gerbino","sequence":"additional","affiliation":[{"name":"Department of Biosciences, Biotechnologies, and Biopharmaceutics University of Bari  70125 Bari Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Geoffrey N.","family":"Hendy","sequence":"additional","affiliation":[{"name":"Department of Medicine Physiology and Human Genetics McGill University  Montreal Quebec Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Torretta","sequence":"additional","affiliation":[{"name":"Department of Biosciences, Biotechnologies, and Biopharmaceutics University of Bari  70125 Bari Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Federica","family":"Rizzo","sequence":"additional","affiliation":[{"name":"Department of Biosciences, Biotechnologies, and Biopharmaceutics University of Bari  70125 Bari Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucantonio","family":"Debellis","sequence":"additional","affiliation":[{"name":"Department of Biosciences, Biotechnologies, and Biopharmaceutics University of Bari  70125 Bari Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppe","family":"Procino","sequence":"additional","affiliation":[{"name":"Department of Biosciences, Biotechnologies, and Biopharmaceutics University of Bari  70125 Bari Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Svelto","sequence":"additional","affiliation":[{"name":"Department of Biosciences, Biotechnologies, and Biopharmaceutics University of Bari  70125 Bari Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2015,3,12]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"crossref","first-page":"C455","DOI":"10.1152\/ajpcell.1996.271.2.C455","article-title":"Na(+)\u2010K+(NH4+)\u20102Cl\u2010 cotransport in medullary thick ascending limb: control by PKA, PKC, and 20\u2010HETE","volume":"271","author":"Amlal H.","year":"1996","journal-title":"Am. 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