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First published online March 14, 2005
Journal of Experimental Biology 208, 1011-1017 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01488

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Nitric oxide synthase in the gill of Atlantic salmon: colocalization with and inhibition of Na⁺,K⁺-ATPase

Lars O. E. Ebbesson^1,*, Christian K. Tipsmark², Bo Holmqvist³, Tom Nilsen¹, Eva Andersson¹, Sigurd O. Stefansson¹ and Steffen S. Madsen²

¹ Department of Biology, University of Bergen, Bergen High Technology Centre, N-5020 Bergen, Norway
² Institute of Biology, University of Southern Denmark, Odense University, Campusvej 55, DK-5230 Odense M, Denmark
³ Department of Pathology, Lund University, Sölvegatan 25, S-221 85 Lund, Sweden

View larger version (145K): [in a new window]   Fig. 1. (A-D) The presence of NO-producing cells (A-C) was compared with that of Na+,K+-ATPase (NKA) -subunit distribution (D) in the gill of Atlantic salmon. nNOS (A) and eNOS (B) immunoreactive cells was mainly restricted to large cells at the base of and along the secondary lamellae (large arrowheads) and in the interlamellar spaces of the primary filaments. Only small cells located deep in the filament were iNOS immunoreactive (C, small arrowheads). (E) NADPHd histochemistry revealed strong staining of large cells similar to the staining with nNOS and eNOS. In addition, a distinct NADPHd punctuate staining (arrows) and band staining (*) were observed on the lamellae (E). Scale bar, 100 µm.

View larger version (187K): [in a new window]   Fig. 2. (A) The expression of Na+,K+-ATPase (NKA) mRNA in the gill was determined using the antisense probe and was found in distinct cells on the primary filament and lamellae (large arrowheads). (B) The sense Na+,K+-ATPase mRNA probe revealed no staining. (C,E) The Na+,K+-ATPase mRNA and protein were colocalized within the same cells (small arrows). (D,F) Double labeling for Na+,K+-ATPase mRNA and nNOS revealed colocalization of these enzymes (arrowheads). Scale bar, 100 µm.

View larger version (182K): [in a new window]   Fig. 3. NADPHd histochemical staining in the gill of salmon parr (A,C) and smolt (B,D). Scale bar, 200 µm (A,B), 100 µm (C,D).