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First published online May 24, 2005
Journal of Experimental Biology 208, 2023-2036 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01611
Functional classification of mitochondrion-rich cells in euryhaline Mozambique tilapia (Oreochromis mossambicus) embryos, by means of triple immunofluorescence staining for Na+/K+-ATPase, Na+/K+/2Cl- cotransporter and CFTR anion channel
1 Department of Anatomy, St Marianna University School of Medicine,
Miyamae-ku, Kawasaki 216-8511, Japan
2 USGS, Conte Anadromous Fish Research Center, Turners Falls, MA 01376,
USA
3 Department of Biology, University of Massachusetts, Amherst, MA 01003,
USA
4 Department of Aquatic Bioscience, Graduate School of Agricultural and Life
Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan
* Author for correspondence (e-mail: j-hiroi{at}marianna-u.ac.jp)
Accepted 16 March 2005
Mozambique tilapia Oreochromis mossambicus embryos were transferred from freshwater to seawater and vice versa, and short-term changes in the localization of three major ion transport proteins, Na+/K+-ATPase, Na+/K+/2Cl- cotransporter (NKCC) and cystic fibrosis transmembrane conductance regulator (CFTR) were examined within mitochondrion-rich cells (MRCs) in the embryonic yolk-sac membrane. Triple-color immunofluorescence staining allowed us to classify MRCs into four types: type I, showing only basolateral Na+/K+-ATPase staining; type II, basolateral Na+/K+-ATPase and apical NKCC; type III, basolateral Na+/K+-ATPase and basolateral NKCC; type IV, basolateral Na+/K+-ATPase, basolateral NKCC and apical CFTR. In freshwater, type-I, type-II and type-III cells were observed. Following transfer from freshwater to seawater, type-IV cells appeared at 12 h and showed a remarkable increase in number between 24 h and 48 h, whereas type-III cells disappeared. When transferred from seawater back to freshwater, type-IV cells decreased and disappeared at 48 h, type-III cells increased, and type-II cells, which were not found in seawater, appeared at 12 h and increased in number thereafter. Type-I cells existed consistently irrespective of salinity changes. These results suggest that type I is an immature MRC, type II is a freshwater-type ion absorptive cell, type III is a dormant type-IV cell and/or an ion absorptive cell (with a different mechanism from type II), and type IV is a seawater-type ion secretory cell. The intracellular localization of the three ion transport proteins in type-IV cells is completely consistent with a widely accepted model for ion secretion by MRCs. A new model for ion absorption is proposed based on type-II cells possessing apical NKCC.
Key words: mitochondrion-rich cell, chloride cell, Na+/K+-ATPase, Na+/K+/2Cl- cotransporter, cystic fibrosis transmembrane conductance regulator, tilapia, Oreochromis mossambicus
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