title: Functional characterization and localization of a gill-specific claudin isoform in Atlantic salmon.
abstract: Claudins are the major determinants of paracellular epithelial permeability in multicellular organisms. In Atlantic salmon (Salmo salar L.), we previously found that mRNA expression of the abundant gill-specific claudin 30 decreases during seawater (SW) acclimation, suggesting that this claudin is associated with remodeling of the epithelium during salinity change. This study investigated localization, protein expression, and function of claudin 30. Confocal microscopy showed that claudin 30 protein was located at cell-cell interfaces in the gill filament in SW- and fresh water (FW)-acclimated salmon, with the same distribution, overall, as the tight junction protein ZO-1. Claudin 30 was located at the apical tight junction interface and in cell membranes deeper in the epithelia. Colocalization with the α-subunit of the Na(+)-K(+)-ATPase was negligible, suggesting limited association with mitochondria-rich cells. Immunoblotting of gill samples showed lower claudin 30 protein expression in SW than FW fish. Retroviral transduction of claudin 30 into Madin-Darby canine kidney cells resulted in a decreased conductance of 19%. The decreased conductance correlated with a decreased permeability of the cell monolayer to monovalent cations, whereas permeability to chloride was unaffected. Confocal microscopy revealed that claudin 30 was expressed in the lateral membrane, as well as in tight junctions of Madin-Darby canine kidney cells, thereby paralleling the findings in the native gill. This study suggests that claudin 30 functions as a cation barrier between pavement cells in the gill and also has a general role in cell-cell adhesion in deeper layers of the epithelium.
authors: Institute of Biology, Univ. of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark. mengelund@biology.sdu.dk
authors: M B
authors: MB
authors: Engelund
authors: A S L
authors: AS
authors: Yu
authors: J
authors: J
authors: Li
authors: S S
authors: SS
authors: Madsen
authors: N J
authors: NJ
authors: Færgeman
authors: C K
authors: CK
authors: Tipsmark
mesh_terms: D000818:Animals; D057167:Claudins / Q000235:genetics / Q000378:metabolism*; D005618:Fresh Water; D005786:Gene Expression Regulation; D005880:Gills / Q000378:metabolism*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D054712:Salinity; D020317:Salmo salar / Q000235:genetics / Q000378:metabolism*; D012623:Seawater; D019108:Tight Junctions / Q000235:genetics / Q000378:metabolism*
chemical_list: D057167:Claudins; D020033:Protein Isoforms