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First published online March 2, 2007
Journal of Experimental Biology 210, 1015-1024 (2007)
Published by The Company of Biologists 2007
doi: 10.1242/jeb.002030
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Variation in salinity tolerance, gill Na+/K+-ATPase, Na+/K+/2Cl– cotransporter and mitochondria-rich cell distribution in three salmonids Salvelinus namaycush, Salvelinus fontinalis and Salmo salar

Junya Hiroi* and Stephen D. McCormick

USGS, Conte Anadromous Fish Research Center, Turners Falls, MA 01376, USA and Department of Biology, University of Massachusetts, Amherst, MA 01003, USA


Figure 1
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  		Fig. 1. Changes in plasma Na+, Cl–, K+ (A–C) and cortisol (D–F) concentrations, and gill Na+/K+-ATPase activity (G–I) in lake trout (A,D,G), brook trout (B,E,H) and Atlantic salmon (C,F,I) following direct transfer from 0 p.p.t. to 30 p.p.t. Each value represents the mean ± s.e.m., N=10 (except for 7 days in A,D,G: N=8; in B,E,H: N=5). Symbols in A–C: circles, Na+; triangles, Cl–; squares, K+. Values with different lowercase letters are significantly different (e.g. a significant difference is observed between `a' and `b', but not between `a' and `a,b', or between `b' and `a,b', P<0.05, Tukey–Kramer test).

 

Figure 2
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  		Fig. 2. Changes in plasma Na+, Cl–, K+ (A–C) and cortisol (D–F) concentrations and gill Na+/K+-ATPase activity (G–I) in lake trout (A,D,G), brook trout (B,E,H) and Atlantic salmon (C,F,I) following gradual transfer from 0 p.p.t. to 10, 20 and 30 p.p.t. Salinities: before day 0, 0 p.p.t.; day 0–7, 10 p.p.t.; day 7–14, 20 p.p.t.; day 14–35, 30 p.p.t. Symbols in A–C: circles, Na+; triangles, Cl–; squares, K+. Each value represents the mean ± s.e.m., N=8. Groups with different lowercase letters are significantly different (P<0.05, Tukey–Kramer test).

 

Figure 3
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  		Fig. 3. Representative western blots of Na+/K+-ATPase (A) and Na+/K+/2Cl– cotransporter (B) from the gills of lake trout, brook trout and Atlantic salmon kept in 0 p.p.t. (FW) and those transferred gradually and kept in 30 p.p.t. for 3 weeks (SW). Molecular mass standards are indicated on the left.

 

Figure 4
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  		Fig. 4. Band intensities of western blots of Na+/K+-ATPase (A) and Na+/K+/2Cl– cotransporter (B) from the gills of lake trout, brook trout and Atlantic salmon kept in 0 p.p.t. (FW) and those transferred gradually and kept in 30 p.p.t. for 3 weeks (SW). Each value represents the mean ± s.e.m., N=4. Asterisks indicate significant differences between FW and SW values (P<0.05, Wilcoxon rank-sum test).

 

Figure 5
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  		Fig. 5. Na+/K+-ATPase (green in A,C,E,G,I,K) and Na+/K+/2Cl– cotransporter (red in B,D,F,H,J,L) immunoreactivity in the gills of lake trout (A–D), brook trout (E–H) and Atlantic salmon (I–L) kept in 0 p.p.t. (freshwater; A,B,E,F,I,J) and those transferred gradually and kept in 30 p.p.t. (seawater) for 3 weeks (C,D,G,H,K,L). The nuclei were counterstained with DAPI (blue). The primary filaments are horizontal, and about three secondary lamellae are seen perpendicular to each filament. Scale bar, 20 µm.

 

Figure 6
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  		Fig. 6. Quantification of Na+/K+-ATPase-immunoreactive mitochondrion-rich cell numbers (A,B) and size (C,D) in the gills of lake trout, brook trout and Atlantic salmon kept in 0 p.p.t. (FW) and those transferred gradually and kept in 30 p.p.t. for 3 weeks (SW). Mitochondria-rich cells on the primary filaments (A,C) and secondary lamellae (B,D) were counted and analyzed separately. Each value represents the mean ± s.e.m., N=4. Asterisks indicate significant differences between FW and SW (P<0.05, Wilcoxon rank-sum test).

 




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