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Fig. 8. Quantification of Na⁺/K⁺-ATPase per chloride cell (CC) and CC area dependence on salinity and acclimation period. (A) Tissue microarray containing 1 mm cores of gill filament from fish acclimated to FW or SW for different times. Bar, 1 mm. (B) Na⁺/K⁺-ATPase antibody combined with PacificBlue-conjugated secondary antibody specifically labels CC. (C) Automatic laser scanning cytometer (LSC) contouring of CC in the same area as shown in A. (D) Quantification of Na⁺/K⁺-ATPase content per CC based on LSC analysis (FW, blue diamonds; 1x SW, green circles; 2.4x SW, red triangles). Na⁺/K⁺-ATPase in CC is significantly but transiently elevated only at 1 week for fish acclimated to 1x SW (P<0.05). In contrast, Na⁺/K⁺-ATPase in CC is significantly elevated at all times measured in fish acclimated to 2.4x SW (P<0.05) (E) CC area based on contouring of Na⁺/K⁺-ATPase fluorescence as shown in B. Although elevated at all times for both 1x and 2.4x SW groups the increase is statistically significant only at 1 week (P<0.05; ^aFW vs. 1x SW, ^bFW vs. 2.4x SW, ^c1x SW vs. 2.4x SW). N=4 animals for each salinity and time.

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