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We performed transcriptomic, molecular and functional studies to identify Cl\u2212 transporters in the seawater-type ionocytes of sea lamprey (Petromyzon marinus). Gill cftr expression was minimal or undetectable in larvae and post-metamorphic juveniles. We identified other Cl\u2212 transporters highly expressed in the gills and\/or upregulated following metamorphosis and further investigated two candidates that stood out in our analysis, a Ca2+-activated Cl\u2212 channel, anoctamin 1 (ano1), and the Clc chloride channel family member 2 (clcn2). Of these, ano1 was expressed 10\u2013100 times more than clcn2 in the gills; moreover, ano1 was upregulated during seawater acclimation, while clcn2 was not. Using an antibody raised against sea lamprey Ano1, we did not detect Ano1 in the gills of larvae, found elevated levels in juveniles and observed a 4-fold increase in juveniles after seawater acclimation. Ano1 was localized to seawater-type branchial ionocytes but, surprisingly, was localized to the basolateral membrane. In vivo pharmacological inhibition experiments demonstrated that a DIDS-sensitive mechanism was critical to the maintenance of osmoregulatory homeostasis in seawater- but not freshwater-acclimated sea lamprey. Taken together, our results provide evidence of a Cftr-independent mechanism for branchial Cl\u2212 secretion in sea lamprey that leverages Ano1-expressing ionocytes. Once further characterized, the Cftr-independent, Ano1-rich ionocytes of sea lamprey could reveal novel strategies for branchial Cl\u2212 secretion, whether by Ano1 or some other Cl\u2212 transporter, not previously known in ionoregulating marine organisms.<\/jats:p>","DOI":"10.1242\/jeb.250110","type":"journal-article","created":{"date-parts":[[2025,2,26]],"date-time":"2025-02-26T03:45:01Z","timestamp":1740541501000},"source":"Crossref","is-referenced-by-count":5,"title":["A Cftr-independent, Ano1-rich seawater-adaptive ionocyte in sea lamprey gills"],"prefix":"10.1242","volume":"228","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2146-9126","authenticated-orcid":false,"given":"Ciaran A.","family":"Shaughnessy","sequence":"first","affiliation":[{"name":"Oklahoma State University 1 Department of Integrative Biology , , Stillwater, OK 74078 ,","place":["USA"]},{"name":"University of Massachusetts 2 Organismic and Evolutionary Biology , , Amherst, MA 01003 ,","place":["USA"]},{"name":"Eastern Ecological Science Center, Conte Research Laboratory 3 US Geological Survey , , Turners Falls, MA 01376 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel J.","family":"Hall","sequence":"additional","affiliation":[{"name":"Eastern Ecological Science Center, Conte Research Laboratory 3 US Geological Survey , , Turners Falls, MA 01376 ,","place":["USA"]},{"name":"University of Massachusetts 4 Department of Biology , , Amherst, MA 01003 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica L.","family":"Norstog","sequence":"additional","affiliation":[{"name":"University of Massachusetts 2 Organismic and Evolutionary Biology , , Amherst, MA 01003 ,","place":["USA"]},{"name":"Eastern Ecological Science Center, Conte Research Laboratory 3 US Geological Survey , , Turners Falls, MA 01376 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andre","family":"Barany","sequence":"additional","affiliation":[{"name":"Eastern Ecological Science Center, Conte Research Laboratory 3 US Geological Survey , , Turners Falls, MA 01376 ,","place":["USA"]},{"name":"Universidad de C\u00e1diz 5 Departamento de Biolog\u00eda , , 11003 C\u00e1diz ,","place":["Spain"]},{"name":"Universidad Complutense 6 Departamento de Gen\u00e9tica, Fisiolog\u00eda y Microbiolog\u00eda , , 28040 Madrid ,","place":["Spain"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amy M.","family":"Regish","sequence":"additional","affiliation":[{"name":"Eastern Ecological Science Center, Conte Research Laboratory 3 US Geological Survey , , Turners Falls, MA 01376 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diogo","family":"Ferreira-Martins","sequence":"additional","affiliation":[{"name":"Eastern Ecological Science Center, Conte Research Laboratory 3 US Geological Survey , , Turners Falls, MA 01376 ,","place":["USA"]},{"name":"University of Massachusetts 4 Department of Biology , , Amherst, MA 01003 ,","place":["USA"]},{"name":"Interdisciplinary Center of Marine and Environmental Research, University of Porto 7 , 4450-208 Matosinhos ,","place":["Portugal"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jason P.","family":"Breves","sequence":"additional","affiliation":[{"name":"Skidmore College 8 Department of Biology , , Saratoga Springs, NY 12866 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa M.","family":"Komoroske","sequence":"additional","affiliation":[{"name":"University of Massachusetts 2 Organismic and Evolutionary Biology , , Amherst, MA 01003 ,","place":["USA"]},{"name":"University of Massachusetts 9 Department of Environmental Conservation , , Amherst, MA 01003 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen D.","family":"McCormick","sequence":"additional","affiliation":[{"name":"Eastern Ecological Science Center, Conte Research Laboratory 3 US Geological Survey , , Turners Falls, MA 01376 ,","place":["USA"]},{"name":"University of Massachusetts 4 Department of Biology , , Amherst, MA 01003 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","published-online":{"date-parts":[[2025,4,2]]},"reference":[{"key":"2025040203590610000_JEB250110C1","doi-asserted-by":"publisher","first-page":"R410","DOI":"10.1152\/ajpregu.00033.2019","article-title":"Osmoregulatory role of the intestine in the sea lamprey (Petromyzon marinus)","volume":"318","author":"Barany","year":"2020","journal-title":"Am. 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