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Fig. 4. The effects of exposure to hypercarbia on the expression of trout
cytoplasmic carbonic anhydrase (tCAc) protein in the gills of rainbow trout
(Oncorhynchus mykiss). An antibody raised against a peptide sequence
specific to tCAc was used to detect tCAc protein expression by (A) western
analysis and (B,C) immunohistochemistry in perfused gills from fish held under
control conditions or exposed to hypercarbia (nominal
PWCO2=6 Torr; 1 Torr=133.3 Pa) for 24 h. In A,
expression of a 
30 kDa protein was significantly greater (one-tailed
Student's t-test, P=0.031) in gills extracted from
hypercarbic trout (N=6) than in those extracted from normocarbic fish
(N=6). Protein expression from the immunoblots presented on the right
was quantified by digital image processing software and depicted in the figure
on the left. In B and C, representative overlay images visualizing tCAc
immunoreactivity (green), 
5 immunoreactivity (red) and nuclei
(4',6'-diamidino-2-phenylindole; blue) are presented for gill
sections from trout exposed to (B) control conditions and (C) hypercarbia for
24 h. Co-localization of tCAc and 5 is indicated in yellow. Scale bars,
20 µm.
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