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First published online October 7, 2005
Journal of Experimental Biology 208, 3851-3858 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01848

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Chloride turnover and ion-transporting activities of yolk-sac preparations (yolk balls) separated from Mozambique tilapia embryos and incubated in freshwater and seawater

Junya Hiroi^1,*, Hiroaki Miyazaki², Fumi Katoh³, Ritsuko Ohtani-Kaneko¹ and Toyoji Kaneko³

¹ Department of Anatomy, St Marianna University School of Medicine, Miyamae, Kawasaki 216-8511, Japan
² Department of Molecular Cell Biology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kamigyo, Kyoto 602-8566, Japan
³ Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo, Tokyo 113-8657, Japan

View larger version (66K): [in a new window]   Fig. 1. Yolk balls prepared from Mozambique tilapia embryos and incubated in freshwater (A) and seawater (B) for 48 h. The yolk sac is completely enclosed by the yolk-sac membrane. The yolk balls slightly swelled in freshwater and shrunk in seawater. Bar, 200 µm.

View larger version (104K): [in a new window]   Fig. 2. Distribution patterns of ion-transporting proteins in mitochondrion-rich cells (MRCs) in the yolk-sac membrane of tilapia yolk balls incubated in freshwater (A–D) and seawater (E–H). The yolk-sac membranes were stained with anti-Na+/K+-ATPase (red; A,E), anti-Na+/K+/2Cl– cotransporter (NKCC) (blue; B,F), and anti-cystic fibrosis transmembrane conductance regulator (CFTR) (green; C,G). (D,H) Merged images of A–C (D) and E–G (H). Arrowheads and arrows indicate apical NKCC staining and apical CFTR staining, respectively. (I, II, III, IV) Functional classification of MRCs is based on distribution patterns of Na+/K+-ATPase, NKCC and CFTR, according to Hiroi et al. (2005). Scale bar, 10 µm.

View larger version (49K): [in a new window]   Fig. 3. Enlarged images of a typical mitochondrion-rich cell in freshwater yolk balls. (A,C,E,G) X–Z optical sections, cut transversely at the horizontal lines indicated in H and I. (B,D,F,H,I) X–Y optical sections, cut transversely at the lines indicated in G. n, nucleus. Scale bar, 10 µm.

View larger version (52K): [in a new window]   Fig. 4. Enlarged images of a typical mitochondrion-rich cell in seawater yolk balls. (A,C,E,G) X–Z optical sections. (B,D,F,H,I) X–Y optical sections. n, nucleus; ac, nucleus of accessory cell. Scale bar, 10 µm.

View larger version (170K): [in a new window]   Fig. 5. Scanning electron micrographs of the yolk-sac membrane of tilapia yolk balls incubated in freshwater (A) and seawater (B) for 48 h. Apical openings (arrows) of mitochondrion-rich cells are located at the boundary of pavement cells possessing microridge structures. Scale bar, 10 µm.