{"id":195013,"date":"2024-08-23T05:31:28","date_gmt":"2024-08-23T10:31:28","guid":{"rendered":"https:\/\/lifeboat.com\/blog\/2024\/08\/gene-expression-and-protein-localization-of-cl%e2%88%92-transporters-slc26a6-and-cftr-in-the-gill-ionocytes-of-rainbow-trout"},"modified":"2024-08-23T05:31:28","modified_gmt":"2024-08-23T10:31:28","slug":"gene-expression-and-protein-localization-of-cl%e2%88%92-transporters-slc26a6-and-cftr-in-the-gill-ionocytes-of-rainbow-trout","status":"publish","type":"post","link":"https:\/\/lifeboat.com\/blog\/2024\/08\/gene-expression-and-protein-localization-of-cl%e2%88%92-transporters-slc26a6-and-cftr-in-the-gill-ionocytes-of-rainbow-trout","title":{"rendered":"Gene expression and protein localization of Cl\u2212 transporters, Slc26a6 and Cftr, in the gill ionocytes of rainbow trout"},"content":{"rendered":"<p><a class=\"aligncenter blog-photo\" href=\"https:\/\/lifeboat.com\/blog.images\/gene-expression-and-protein-localization-of-cle28892-transporters-slc26a6-and-cftr-in-the-gill-ionocytes-of-rainbow-trout.jpg\"><\/a><\/p>\n<p>In rainbow trout, classification of ionocytes based on the expressed transporters is still in progress. The Nhe3-positive ionocytes expressing basolateral Nka and Nkcc1 and apical Nhe3b have been found in both freshwater-and seawater-acclimated rainbow trout. Colocalization of an Rh glycoprotein (Rhcg1, ammonium transporter) and Nhe3b at the apical membrane was also observed, suggesting ammonia-dependent Na<sup>+<\/sup> uptake by Nhe3-positive ionocytes (Hiroi and McCormick <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2012\" title=\"Hiroi J, McCormick SD (2012) New insights into gill ionocyte and ion transporter function in euryhaline and diadromous fish. Respir Physiol Neurobiol 184:257&ndash;268\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR11\" id=\"ref-link-section-d91905136e553\">2012<\/a>). The Nhe3-negative ionocytes, which also lack Nkcc1, are observed mainly in freshwater (Katoh <i>et al.<\/i> <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2008\" title=\"Katoh F, Cozzi RR, Marshall WS, Goss GG (2008) Distinct Na+\/K+\/2Cl-cotransporter localization in kidneys and gills of two euryhaline species, rainbow trout and killifish. Cell Tissue Res 334:265&ndash;281\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR19\" id=\"ref-link-section-d91905136e556\">2008<\/a>; Hiroi and McCormick <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2012\" title=\"Hiroi J, McCormick SD (2012) New insights into gill ionocyte and ion transporter function in euryhaline and diadromous fish. Respir Physiol Neurobiol 184:257&ndash;268\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR11\" id=\"ref-link-section-d91905136e559\">2012<\/a>). Ncc2, the apical Cl<sup>\u2212<\/sup> pathway in tilapia type-II ionocytes, is thought to be absent in the gill of salmonids (Hiroi and McCormick <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2012\" title=\"Hiroi J, McCormick SD (2012) New insights into gill ionocyte and ion transporter function in euryhaline and diadromous fish. Respir Physiol Neurobiol 184:257&ndash;268\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR11\" id=\"ref-link-section-d91905136e565\">2012<\/a>). The Nhe3-positive ionocytes showed basolateral Nka and Nkcc1 both in freshwater and seawater, suggesting that Nhe3-positive ionocytes are analogous to tilapia types-III and-IV and could be equipped with apical Cftr in seawater (Hiroi and McCormick <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2012\" title=\"Hiroi J, McCormick SD (2012) New insights into gill ionocyte and ion transporter function in euryhaline and diadromous fish. Respir Physiol Neurobiol 184:257&ndash;268\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR11\" id=\"ref-link-section-d91905136e568\">2012<\/a>; Takei <i>et al.<\/i> <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2014\" title=\"Takei Y, Hiroi J, Takahashi H, Sakamoto T (2014) Diverse mechanisms for body fluid regulation in teleost fishes. Am J Physiol Regul Integr Comp Physiol 307:R778&ndash;R792\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR35\" id=\"ref-link-section-d91905136e571\">2014<\/a>). However, localization of Cftr proteins by immunohistochemistry has not been successful in salmonids even with homologous antibodies (Takei <i>et al.<\/i> <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2014\" title=\"Takei Y, Hiroi J, Takahashi H, Sakamoto T (2014) Diverse mechanisms for body fluid regulation in teleost fishes. Am J Physiol Regul Integr Comp Physiol 307:R778&ndash;R792\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR35\" id=\"ref-link-section-d91905136e574\">2014<\/a>). The mRNA of <i>slc26a6<\/i> has been reported to be highly expressed in the gills of freshwater-acclimated rainbow trout (Boyle <i>et al.<\/i> <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2015\" title=\"Boyle D, Clifford AM, Orr E, Chamot D, Goss GG (2015) Mechanisms of Cl\u2212 uptake in rainbow trout: cloning and expression of slc26a6, a prospective Cl\u2212\/HCO3\u2212 exchanger. Comp Biochem Physiol A Mol Integr Physiol 180:43&ndash;50\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR2\" id=\"ref-link-section-d91905136e580\">2015<\/a>; Leguen <i>et al.<\/i> <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2015\" title=\"Leguen I, Le Cam A, Montfort J, Peron S, Fautrel A (2015) Transcriptomic analysis of trout gill ionocytes in fresh water and sea water using laser capture microdissection combined with microarray analysis. PLoS ONE 10:e0139938\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR24\" id=\"ref-link-section-d91905136e584\">2015<\/a>) and it is very likely that this transporter is responsible for the uptake of Cl<sup>\u2212<\/sup> in freshwater, but detailed localization of this protein in the gills has not been elucidated. In short, the molecules responsible for the Cl<sup>\u2212<\/sup> transport across the apical membrane have not been identified in both freshwater-and seawater-acclimated rainbow trout.<\/p>\n<p>Salmonids possess two <i>cftr<\/i> genes, <i>cftr1<\/i> and <i>cftr2<\/i> (Chen <i>et al.<\/i> <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2001\" title=\"Chen JM, Cutler C, Jacques C, B\u0153uf G, Denamur E, Lecointre G, Mercier B, Cramb G, F\u00e9rec C (2001) A combined analysis of the cystic fibrosis transmembrane conductance regulator: implications for structure and disease models. Mol Biol Evol 18:1771&ndash;1788\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR4\" id=\"ref-link-section-d91905136e603\">2001<\/a>), and it has been reported that the expression level of both genes increases in the gill of chum salmon <i>Oncorhynchus keta<\/i> after the transfer to seawater during the juvenile stage (Wong <i>et al.<\/i> <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2019\" title=\"Wong MKS, Nobata S, Hyodo S (2019) Enhanced osmoregulatory ability marks the smoltification period in developing chum salmon (Oncorhynchus keta). Comp Biochem Physiol A Mol Integr Physiol 238:110565\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR40\" id=\"ref-link-section-d91905136e610\">2019<\/a>). Expression of <i>cftr1<\/i> in the gills increased also in rainbow trout after seawater transfer (Gerber <i>et al.<\/i> <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2018\" title=\"Gerber L, Jensen FB, Madsen SS (2018) Dynamic changes in nitric oxide synthase expression are involved in seawater acclimation of rainbow trout Oncorhynchus mykiss. Am J Physiol Regul Integr Comp Physiol 314:R552&ndash;R562\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR8\" id=\"ref-link-section-d91905136e616\">2018<\/a>). On the other hand, dietary salt loading reduced <i>cftr2<\/i> expression in the gill of rainbow trout in freshwater (Kolosov and Kelly <a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 2016\" title=\"Kolosov D, Kelly SP (2016) Dietary salt loading and ion-poor water exposure provide insight into the molecular physiology of the rainbow trout gill epithelium tight junction complex. J Comp Physiol B 186:739&ndash;757\" href=\"https:\/\/link.springer.com\/article\/10.1007\/s12562-024-01809-7#ref-CR22\" id=\"ref-link-section-d91905136e622\">2016<\/a>). At this time, it is not clear which of these two molecules is mainly responsible for hypo-osmoregulatory Cl<sup>\u2212<\/sup> secretion in the gills of salmonids.<\/p>\n<p>The objective of the present study is to examine molecules responsible for the active transport of Cl<sup>\u2212<\/sup> in gill ionocytes of rainbow trout. To achieve this goal, we conducted tissue distribution analyses on the expression of <i>slc26a6<\/i>, <i>cftr1<\/i>, and <i>cftr2<\/i> in rainbow trout acclimated to freshwater or seawater. Time-course changes in the expression of these genes were also examined during seawater transfer. We localized these proteins in the gill filaments of rainbow trout acclimated to freshwater or seawater by whole-mount immunohistochemistry.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>In rainbow trout, classification of ionocytes based on the expressed transporters is still in progress. The Nhe3-positive ionocytes expressing basolateral Nka and Nkcc1 and apical Nhe3b have been found in both freshwater-and seawater-acclimated rainbow trout. Colocalization of an Rh glycoprotein (Rhcg1, ammonium transporter) and Nhe3b at the apical membrane was also observed, suggesting ammonia-dependent Na+ [\u2026]<\/p>\n","protected":false},"author":661,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[19],"tags":[],"class_list":["post-195013","post","type-post","status-publish","format-standard","hentry","category-chemistry"],"_links":{"self":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/195013","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/users\/661"}],"replies":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/comments?post=195013"}],"version-history":[{"count":0,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/195013\/revisions"}],"wp:attachment":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/media?parent=195013"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/categories?post=195013"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/tags?post=195013"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}