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First published online January 31, 2006
Journal of Experimental Biology 209, 599-609 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02059
Microtubule-dependent relocation of branchial V-H+-ATPase to the basolateral membrane in the Pacific spiny dogfish (Squalus acanthias): a role in base secretion
Dept of Biological Sciences, University of Alberta, Edmonton, Alberta, T5G 2E9, Canada and Bamfield Marine Research Centre, Bamfield, British Columbia, V0R 1B0, Canada
* Author for correspondence (e-mail: martint{at}ualberta.ca)
Accepted 22 December 2005
We have previously shown that continuous intravenous infusion of
NaHCO3 for 24 h (
1000 µmol kg-1 h-1)
results in the relocation of V-H+-ATPase from the cytoplasm to the
basolateral membrane in the gills of the Pacific dogfish. To further
investigate this putative base-secretive process we performed similar
experiments with the addition of colchicine, an inhibitor of
cytoskeleton-dependent cellular trafficking processes. Blood pH and plasma
total CO2 were significantly higher in the colchicines-treated,
HCO3--infused fish compared with fish infused with
HCO3- alone. The effect of colchicine was highest after
24 h of infusion (8.33±0.06 vs 8.02±0.03 pH units,
15.72±3.29 vs 6.74±1.34 mmol CO2
l-1, N=5). Immunohistochemistry and western blotting
confirmed that colchicine blocked the transit of V-H+-ATPase to the
basolateral membrane. Furthermore, western blotting analyses from whole gill
and cell membrane samples suggest that the short-term (6 h) response to
alkaline stress consists of relocation of V-H+-ATPases already
present in the cell to the basolateral membrane, while in the longer term (24
h) there is both relocation of preexistent enzyme and upregulation in the
synthesis of new units. Our results strongly suggest that cellular relocation
of V-H+-ATPase is necessary for enhanced
HCO3- secretion across the gills of the Pacific
dogfish.
Key words: fish, dogfish, Squalus acanthias, gill, acid base regulation, H+-ATPase, base infusion, alkalosis, colchicine, basolateral H+-ATPase, microtubule, vesicular trafficking.
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