{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T23:15:04Z","timestamp":1783120504384,"version":"3.54.6"},"reference-count":75,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2025,12,23]],"date-time":"2025-12-23T00:00:00Z","timestamp":1766448000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,12,23]],"date-time":"2025-12-23T00:00:00Z","timestamp":1766448000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["physoc.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Physiology"],"published-print":{"date-parts":[[2026,2]]},"abstract":"\n Abstract<\/jats:title>\n \n CO\n 2<\/jats:sub>\n diffusion across plasma membranes depends on both membrane CO\n 2<\/jats:sub>\n permeability () and the transmembrane CO\n 2<\/jats:sub>\n concentration gradient (\u0394[CO\n 2<\/jats:sub>\n ]) \u2013 Fick's law. Human aquaporin\u20105 (hAQP5) enhances CO\n 2<\/jats:sub>\n diffusion by increasing , whereas carbonic anhydrases (CAs) do so by enhancing CO\n 2<\/jats:sub>\n consumption\/production and thus \u0394[CO\n 2<\/jats:sub>\n ]. Here we systematically assess functional interactions among a gas channel and intra\u2010\/extracellular CAs. On Day 1 we inject\n Xenopus<\/jats:italic>\n oocytes with cRNA encoding hAQP5 (control: H\n 2<\/jats:sub>\n O). On Day 4 we inject hCA\u00a0II protein in \u2018Tris\u2019 buffer (control: \u2018Tris\u2019). We assess CO\n 2<\/jats:sub>\n fluxes by introducing extracellular 1.5% CO\n 2<\/jats:sub>\n \/10 mM HCO\n 3<\/jats:sub>\n \u2212<\/jats:sup>\n and using microelectrodes to measure (1) the maximal increase of extracellular surface pH (\u0394pH\n S<\/jats:sub>\n ), (2) the maximal rate of pH\n S<\/jats:sub>\n relaxation (dpH\n S<\/jats:sub>\n \/dt)\n Max<\/jats:sub>\n and (3) the maximal rate of intracellular pH decrease (dpH\n i<\/jats:sub>\n \/dt)\n Max<\/jats:sub>\n . By itself hCA\u00a0II minimally increases \u0394pH\n S<\/jats:sub>\n \u2013 measured on the side of the membrane opposite to the added cytosolic CA (CA\n i<\/jats:sub>\n ) \u2013 even at our highest doses (100\u00a0ng\/oocyte). However hAQP5 alone triples \u0394pH\n S<\/jats:sub>\n , an effect further doubled by increasing hCA\u00a0II. By itself bovine erythrocyte CA (bCA) in the extracellular fluid doubles (dpH\n i<\/jats:sub>\n \/dt)\n Max<\/jats:sub>\n magnitude \u2013 measured on the side of the membrane opposite to the added extracellular CA (CA\n o<\/jats:sub>\n ) \u2013 an effect further doubled by hAQP5. Our pH measurements (1) confirm synergy between CA\n o<\/jats:sub>\n and CA\n i<\/jats:sub>\n ; establish synergy between hAQP5 and both (2) CA\n o<\/jats:sub>\n and (3) CA\n i<\/jats:sub>\n ; and (4) show that the ability of CA\n i<\/jats:sub>\n to enhance \u0394pH\n S<\/jats:sub>\n is a useful tool for assessing the CO\n 2<\/jats:sub>\n permeability of membrane proteins (e.g. hAQP5).\n \n \n image<\/jats:alt-text>\n <\/jats:graphic>\n <\/jats:boxed-text>\n <\/jats:p>\n <\/jats:sec>\n \n Key points<\/jats:title>\n \n \n \n \n According to Fick's law transmembrane CO\n 2<\/jats:sub>\n flux () is the product of membrane permeability () and transmembrane concentration gradient (\u0394[CO\n 2<\/jats:sub>\n ]): \u00a0=\u00a0\u00d7\u0394[CO\n 2<\/jats:sub>\n ]. Previous work separately showed that (1) human aquaporin\u20105 (hAQP5) enhances , and (2) intracellular and (3) extracellular carbonic anhydrases (CAs) enhance (\u0394[CO\n 2<\/jats:sub>\n ]) by consuming accumulated or replenishing lost CO\n 2<\/jats:sub>\n . We now examine interactions among #1\u2013#3.\n <\/jats:p>\n <\/jats:list-item>\n \n \n We assess CO\n 2<\/jats:sub>\n fluxes \u2013 produced by addition\/removal of extracellular CO\n 2<\/jats:sub>\n \/HCO\n 3<\/jats:sub>\n \u2212<\/jats:sup>\n \u2013 using microelectrodes to monitor extracellular surface pH (pH\n S<\/jats:sub>\n ) and intracellular pH (pH\n i<\/jats:sub>\n ) of\n Xenopus<\/jats:italic>\n oocytes heterologously expressing hAQP5, injected with human CA\u00a0II (hCA\u00a0II), and\/or exposed to extracellular bovine CA (bCA).\n <\/jats:p>\n <\/jats:list-item>\n \n \n Enhancing effects on CO\n 2<\/jats:sub>\n fluxes are synergistic among hAQP5, hCA\u00a0II and bCA, any of which can become rate\u2010limiting, depending on the status of the other two.\n <\/jats:p>\n <\/jats:list-item>\n \n \n CO\n 2<\/jats:sub>\n \/HCO\n 3<\/jats:sub>\n \u2212<\/jats:sup>\n addition transiently increases pH\n S<\/jats:sub>\n (\u0394pH\n S<\/jats:sub>\n ), hCA\u00a0II augments \u0394pH\n S<\/jats:sub>\n (\u0394\u0394pH\n S<\/jats:sub>\n ) and hAQP5 enhances \u0394\u0394pH\n S<\/jats:sub>\n (\u0394\u0394\u0394pH\n S<\/jats:sub>\n ) \u2013 a novel tool to assess potential CO\n 2<\/jats:sub>\n channels.\n <\/jats:p>\n <\/jats:list-item>\n <\/jats:list>\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1113\/jp289145","type":"journal-article","created":{"date-parts":[[2025,12,23]],"date-time":"2025-12-23T15:01:15Z","timestamp":1766502075000},"page":"1174-1234","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Synergistic roles of aquaporin 5 and intra\u2010 and extracellular carbonic anhydrases in promoting CO\n 2<\/sub>\n diffusion across the\n Xenopus<\/i>\n oocyte plasma membrane"],"prefix":"10.1113","volume":"604","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-4857-4650","authenticated-orcid":false,"given":"Deng\u2010Ke","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Physiology and Biophysics Case Western Reserve University School of Medicine Cleveland Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8519-6991","authenticated-orcid":false,"given":"Fraser J.","family":"Moss","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics Case Western Reserve University School of Medicine Cleveland Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4741-7287","authenticated-orcid":false,"given":"Walter F.","family":"Boron","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics Case Western Reserve University School of Medicine Cleveland Ohio USA"},{"name":"Department of Medicine Case Western Reserve University School of Medicine Cleveland Ohio USA"},{"name":"Department of Biochemistry Case Western Reserve University School of Medicine Cleveland Ohio 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