{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T17:17:51Z","timestamp":1790702271465,"version":"4.1.0"},"reference-count":51,"publisher":"American Physiological Society","issue":"3","funder":[{"DOI":"10.13039\/100000057","name":"HHS | NIH | National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["T32GM099608"],"award-info":[{"award-number":["T32GM099608"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100015627","name":"Quest Diagnostics","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100015627","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.physiology.org"],"crossmark-restriction":true},"short-container-title":["American Journal of Physiology-Cell Physiology"],"published-print":{"date-parts":[[2022,9,1]]},"abstract":"<jats:p> Red cell volume is a major determinant of HbS concentration in sickle cell disease. Cellular deoxy-HbS concentration determines the delay time, the interval between HbS deoxygenation and deoxy-HbS polymerization. Major membrane transporter protein determinants of sickle red cell volume include the SLC12\/KCC K-Cl cotransporters KCC3\/SLC12A6 and KCC1\/SLC12A4, and the KCNN4\/KCa3.1 Ca<jats:sup>2+<\/jats:sup>-activated K<jats:sup>+<\/jats:sup> channel (Gardos channel). Among standard inhibitors of KCC-mediated K-Cl cotransport, only [(dihydroindenyl)oxy]acetic acid (DIOA) has been reported to lack inhibitory activity against the related bumetanide-sensitive erythroid Na-K-2Cl cotransporter NKCC1\/SLC12A2. DIOA has been often used to inhibit K-Cl cotransport when studying the expression and regulation of other K<jats:sup>+<\/jats:sup> transporters and K<jats:sup>+<\/jats:sup> channels. We report here that DIOA at concentrations routinely used to inhibit K-Cl cotransport can also abrogate activity of the KCNN4\/KCa3.1 Gardos channel in human and mouse red cells and in human sickle red cells. DIOA inhibition of A23187-stimulated erythroid K<jats:sup>+<\/jats:sup> uptake (Gardos channel activity) was chloride-independent and persisted in mouse red cells genetically devoid of the principal K-Cl cotransporters KCC3 and KCC1. DIOA also inhibited YODA1-stimulated, chloride-independent erythroid K<jats:sup>+<\/jats:sup> uptake. In contrast, DIOA exhibited no inhibitory effect on K<jats:sup>+<\/jats:sup> influx into A23187-treated red cells of Kcnn4<jats:sup>\u2212\/\u2212<\/jats:sup> mice. DIOA inhibition of human KCa3.1 was validated (IC<jats:sub>50<\/jats:sub> 42 \u00b5M) by whole cell patch clamp in HEK-293 cells. RosettaLigand docking experiments identified a potential binding site for DIOA in the fenestration region of human KCa3.1. We conclude that DIOA at concentrations routinely used to inhibit K-Cl cotransport can also block the KCNN4\/KCa3.1 Gardos channel in normal and sickle red cells. <\/jats:p>","DOI":"10.1152\/ajpcell.00240.2022","type":"journal-article","created":{"date-parts":[[2022,7,18]],"date-time":"2022-07-18T11:12:22Z","timestamp":1658142742000},"page":"C694-C705","update-policy":"https:\/\/doi.org\/10.1152\/aps-crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["The erythroid K-Cl cotransport inhibitor [(dihydroindenyl)oxy]acetic acid blocks erythroid Ca<sup>2+<\/sup>-activated K<sup>+<\/sup> channel KCNN4"],"prefix":"10.1152","volume":"323","author":[{"given":"Alicia","family":"Rivera","sequence":"first","affiliation":[{"name":"Division of Nephrology and Vascular Biology Research Center, Beth Israel Deaconess Medical Center, Boston, Massachusetts"},{"name":"Department of Medicine, Harvard Medical School, Boston, Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joshua A.","family":"Nasburg","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, School of Medicine, University of California, Davis, California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heesung","family":"Shim","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, School of Medicine, University of California, Davis, California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Boris E.","family":"Shmukler","sequence":"additional","affiliation":[{"name":"Division of Nephrology and Vascular Biology Research Center, Beth Israel Deaconess Medical Center, Boston, Massachusetts"},{"name":"Department of Medicine, Harvard Medical School, Boston, Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jason","family":"Kitten","sequence":"additional","affiliation":[{"name":"Meta Isotopes LLC, Lubbock, Texas"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jay G.","family":"Wohlgemuth","sequence":"additional","affiliation":[{"name":"Quest Diagnostics, Secaucus, New Jersey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffrey S.","family":"Dlott","sequence":"additional","affiliation":[{"name":"Quest Diagnostics, Secaucus, New Jersey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L. 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