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First published online February 6, 2004
Journal of Experimental Biology 207, 905-912 (2004)
Published by The Company of Biologists 2004
doi: 10.1242/jeb.00844

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Seawater acclimation causes independent alterations in Na⁺/K⁺- and H⁺-ATPase activity in isolated mitochondria-rich cell subtypes of the rainbow trout gill

Guy S. Hawkings, Fernando Galvez^* and Greg G. Goss

Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada T6E 4W1

View larger version (12K): [in a new window]   Fig. 1. Ouabain-sensitive Na+/K+-ATPase activity of control and seawater-adapted fish in a total cell fraction [i.e. cells collected from combined PVC/MR (pavement cell/mitochondria-rich) fraction, the 1.03–1.09 g ml–1 Percoll interface]. Both partial-strength seawater (10) and full-strength seawater (30)-adapted fish were used for testing. Significant differences are indicated by different letters. Values represent means ± S.E.M. (N=5 fish).

View larger version (11K): [in a new window]   Fig. 2. Bafilomycin (50 nmol l–1)-sensitive H+-ATPase activity in the total cell fraction [i.e. cells collected from combined PVC/MR (pavement cell/mitochondria-rich) fraction, the 1.03–1.09 g ml–1 Percoll interface]. Freshwater control fish compared with both 10 and 30 seawater-adapted fish. Significant differences are indicated by different letters. Values represent means ± S.E.M. (N=4–6 fish).

View larger version (66K): [in a new window]   Fig. 3. Differential interference contrast (DIC) and fluorescence images of dispersed gill cells from freshwater rainbow trout (A–C) and seawater-adapted (30) trout (D–F). Cells were obtained from the mitochondria-rich (MR) 1.05–1.09 g ml–1 Percoll layer and stained with Mitotracker Green-FM (100 nmol l–1, 20 min; B,E) or labelled with PNA Alexa fluor 594 (40 µg ml–1; C,F) for identification of MR and PNA+ cell types, respectively. All pictures in A–C and D–F are of the same field of view. Scale bar, 10 µm.

View larger version (16K): [in a new window]   Fig. 4. Measured sizes of cells in freshwater and seawater-adapted trout. Cell sizes measured by calibrated measuring tool in viewing software from differential interference contrast (DIC) images taken for seawater adaptation experiments. Values represent means ± S.E.M. (N=30–40 cells for each population).

View larger version (14K): [in a new window]   Fig. 5. Relative percentage of PNA– and PNA+ cell types in populations of gill cells isolated from the MR 1.05–1.09 g ml–1 Percoll interface. Control fish (freshwater) are compared with seawater-adapted fish at partial-strength (10) and full-strength seawater (30). PNA+ and PNA– cells counted under 100x objective of fluorescence microscope with a minimum of five fields of view counted for each freshwater and seawater fish. Values represent means ± S.E.M. (N=6 fish for each treatment group).

View larger version (13K): [in a new window]   Fig. 6. Ouabain-sensitive Na+/K+-ATPase activity of control and seawater-adapted fish in the PNA– and PNA+ cell fractions. Both partial-strength seawater (10) and full-strength seawater (30)-adapted fish used for testing. Values represent means ± S.E.M. (N=5–6 fish).

View larger version (13K): [in a new window]   Fig. 7. Bafilomycin (50 nmol l–1)-sensitive H+-ATPase activity in the PNA+ and PNA– gill cell fractions. Freshwater control fish compared with both 10 and 30 seawater-adapted fish. Values represent means ± S.E.M. (N=3–5 fish).