title: Multiplicity of expression of Na+,K+-ATPase {alpha}-subunit isoforms in the gill of Atlantic salmon (Salmo salar): cellular localisation and absolute quantification in response to salinity change.
abstract: The ability to reverse the net direction of gill ion transport in response to a salinity change is critical for euryhaline teleosts and involves a complex cellular and molecular remodelling of the gill epithelium. The present study aimed to clarify the cellular localisation and exact quantitative inter-relationship of Na(+),K(+)-ATPase alpha- and beta-subunit transcripts in Atlantic salmon gill during salinity change. The combined expression level of all alpha-isoforms in the gill increased by 100% after freshwater (FW) to seawater (SW) transfer. The alpha(1a) and alpha(1b) isoforms were both in the range 1-6 amol 20 ng(-1) total RNA; alpha(1a) decreased and alpha(1b) increased after SW-transfer, their ratio changing from 5:1 in FW to 0.26:1 in SW. The alpha(1c) and alpha(3) levels were 10- and 100-fold lower, respectively. The beta(1)-subunit mRNA level was 0.1-0.3 amol 20 ng(-1) total RNA, thus much lower than the sum of alpha-subunits. Even though increasing 3-fold after SW-transfer, beta-subunit availability may still limit functional pump synthesis. The mRNAs of the predominant alpha(1a) and alpha(1b) isoforms were localised by in situ hybridisation in specific gill cells of both FW and SW salmon. Labelling occurred mainly in presumed chloride cells and cells deep in the filament but occasionally also on lamellae. Overall, the salinity-induced variation in labelling pattern and intensity matched the quantification data. In conclusion, the predominant switching of Na(+),K(+)-ATPase alpha-subunit isoform mRNA during salinity acclimation reflects a marked remodelling of mitochondrion-rich cells (MRCs) in the gill and probably tuning of the pump performance to accomplish a net reversal of gill ion transport in hypo- and hypertonic environments.
authors.affiliation: Institute of Biology, University of Southern Denmark, 5230 Odense M, Denmark. steffen@biology.sdu.dk
authors.forename: Steffen S
authors.initials: SS
authors.lastname: Madsen
authors.forename: Pia
authors.initials: P
authors.lastname: Kiilerich
authors.forename: Christian K
authors.initials: CK
authors.lastname: Tipsmark
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D003718:Denmark; D004777:Environment; D005880:Gills / Q000378:metabolism*; D017403:In Situ Hybridization; D007527:Isoenzymes / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020317:Salmo salar / Q000378:metabolism*; D012965:Sodium Chloride / Q000032:analysis*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*; D014867:Water / Q000737:chemistry*
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D007527:Isoenzymes; D014867:Water; D012965:Sodium Chloride; D000254:Sodium-Potassium-Exchanging ATPase