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First published online October 10, 2003
Short-term transformation and long-term replacement of branchial chloride cells in killifish transferred from seawater to freshwater, revealed by morphofunctional observations and a newly established `time-differential double fluorescent staining' technique
Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan
* Author for correspondence (e-mail: fkatoh{at}marine.fs.a.u-tokyo.ac.jp)
Accepted 11 August 2003
Short- and long-term responses to direct transfer from seawater to freshwater were examined in gill chloride cells of killifish, which developed distinct freshwater- and seawater-type chloride cells in the respective environments. In a short-term response within 24 h after transfer, seawater-type chloride cells forming a pit structure on the apical surface were transformed into freshwater-type cells equipped with developed microvilli on the flat or projecting apical membrane, via the intermediate type. The transformation process was accompanied by the disappearance of apically located Cl- channel (cystic fibrosis transmembrane conductance regulator) and neighboring accessory cells. Chloride cell replacement was also examined as a long-term adaptation to freshwater transfer, using a newly established `time-differential double fluorescent staining (TDS)' technique. In the TDS technique, in vivo labeling of chloride cells was performed on two separate days, using two distinguishable mitochondria-specific fluorescent probes. For 3 days after freshwater transfer, 14.7% of seawater-type cells were replaced with newly differentiated freshwater-type cells, whereas these ratios of chloride cell replacement were much lower (1.2% and 1.8%) in seawater- and freshwater-maintained groups, respectively. In consequence, following direct transfer of killifish from seawater to freshwater, seawater-type chloride cells were transformed morphologically and functionally into freshwater-type cells as a short-term response, followed by the promotion of chloride cell replacement as a long-term response.
Key words: branchial chloride cell, killifish, Fundulus heteroclitus, time-differential double fluorescent staining
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