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First published online November 10, 2003
Influence of salinity on the localization of Na+/K+-ATPase, Na+/K+/2Cl- cotransporter (NKCC) and CFTR anion channel in chloride cells of the Hawaiian goby (Stenogobius hawaiiensis)
1 USGS, Leetown Science Center, Conte Anadromous Fish Research Center,
Turners Falls, MA 01370, USA
2 Department of Biology, University of Massachusetts, Amherst, MA 01003,
USA
3 Fish Endocrinology Laboratory, Department of Zoology/Zoophysiology,
Göteborg University, Box 463, S405 30 Göteborg, Sweden
4 Marine Biology Program, Florida International University, 3000 NE 151st
St, North Miami, FL 33181, USA
* Author for correspondence (e-mail: stephen_mccormick{at}usgs.gov)
Accepted 9 September 2003
Na+/K+-ATPase,
Na+/K+/2Cl- cotransporter (NKCC) and cystic
fibrosis transmembrane conductance regulator (CFTR) are the three major
transport proteins thought to be involved in chloride secretion in teleost
fish. If this is the case, the levels of these transporters should be high in
chloride cells of seawater-acclimated fish. We therefore examined the
influence of salinity on immunolocalization of
Na+/K+-ATPase, NKCC and CFTR in the gills of the
Hawaiian goby (Stenogobius hawaiiensis). Fish were acclimated to
freshwater and 20
and 30 seawater for 10 days.
Na+/K+-ATPase and NKCC were localized specifically to
chloride cells and stained throughout most of the cell except for the nucleus
and the most apical region, indicating a basolateral/tubular distribution. All
Na+/K+-ATPase-positive chloride cells were also positive
for NKCC in all salinities. Salinity caused a slight increase in chloride cell
number and size and a slight decrease in staining intensity for
Na+/K+-ATPase and NKCC, but the basic pattern of
localization was not altered. Gill Na+/K+-ATPase
activity was also not affected by salinity. CFTR was localized to the apical
surface of chloride cells, and only cells staining positive for
Na+/K+-ATPase were CFTR-positive. CFTR-positive cells
greatly increased in number (5-fold), area stained (53%) and intensity (29%)
after seawater acclimation. In freshwater, CFTR immunoreactivity was light and
occurred over a broad apical surface on chloride cells, whereas in seawater
there was intense immunoreactivity around the apical pit (which was often
punctate in appearance) and a light subapical staining. The results indicate
that Na+/K+-ATPase, NKCC and CFTR are all present in
chloride cells and support current models that all three are responsible for
chloride secretion by chloride cells of teleost fish.
Key words: Na+/K+-ATPase, Na+/K+/2Cl- cotransporter, NKCC, CFTR, teleost fish, chloride secretion, salinity, Stenogobius hawaiiensis, goby
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