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First published online November 10, 2003

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Influence of salinity on the localization of Na⁺/K⁺-ATPase, Na⁺/K⁺/2Cl^- cotransporter (NKCC) and CFTR anion channel in chloride cells of the Hawaiian goby (Stenogobius hawaiiensis)

Stephen D. McCormick^1,2,*, Kristina Sundell³, Björn Thrandur Björnsson³, Christopher L. Brown⁴ and Junya Hiroi^1,2,

¹ USGS, Leetown Science Center, Conte Anadromous Fish Research Center, Turners Falls, MA 01370, USA
² Department of Biology, University of Massachusetts, Amherst, MA 01003, USA
³ Fish Endocrinology Laboratory, Department of Zoology/Zoophysiology, Göteborg University, Box 463, S405 30 Göteborg, Sweden
⁴ Marine Biology Program, Florida International University, 3000 NE 151st St, North Miami, FL 33181, USA

View larger version (106K): [in a new window]   Fig. 1. Na+/K+-ATPase (green) and Na+/K+/2Cl- cotransporter (NKCC; red) immunoreactivity in gills of Hawaiian goby acclimated to freshwater (A–C) and 30 seawater (D–F). Double-staining was performed on the same sections, and the images combined in C and F. Images C and F were merged (from A,B and D,E, respectively) to highlight the colocalization of Na+/K+-ATPase and NKCC1 (red + green is yellow-orange). The width of each image is 207 µm.

View larger version (77K): [in a new window]   Fig. 2. Na+/K+-ATPase (green) and cystic fibrosis transmembrane conductance regulator (CFTR; red) immunoreactivity in gills of Hawaiian goby acclimated to seawater (A). Image B contains only CFTR staining (white). Scale bar, 12 µm.

View larger version (31K): [in a new window]   Fig. 3. Quantification of Na+/K+-ATPase-immunoreactive (left panels) and cystic fibrosis transmembrane conductance regulator (CFTR; right panels)-immunoreactive chloride cell number, size and staining intensity. Chloride cells on the primary filament (1°) and secondary lamellae (2°) were counted and analyzed separately. Groups with different letters were significantly different from one another (P<0.05, Student–Newman–Keuls test). Values are expressed as means ± S.E.M. At least 50 cells per individual and five individuals in each treatment group were measured.

View larger version (114K): [in a new window]   Fig. 4. Na+/K+-ATPase (green) and cystic fibrosis transmembrane conductance regulator (CFTR; red) immunoreactivity in gills of Hawaiian goby acclimated to freshwater (A–C) and 30 seawater (D–F). Double-staining was performed on the same sections, and the images combined in C and F. Images C and F were overlaid (from A,B and D,E, respectively) to highlight the colocalization of Na+/K+-ATPase and CFTR. The width of each image is 207 µm.

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