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Daily variations of endolymph composition: relationship with the otolith calcification process in trout

G. Borelli1, M. E. Guibbolini1, N. Mayer-Gostan1, F. Priouzeau1, H. De Pontual2, D. Allemand1, S. Puverel3, E. Tambutte3 and P. Payan1,*

1 Laboratoire Réponse des Organismes aux Stress Environnementaux, UMR INRA-UNSA 1112, Université de Nice-Sophia Antipolis, Faculté des Sciences, BP 71, 06108 Nice Cedex 2, France
2 IFREMER, DRV, RH, Laboratoire de Sclérochronologie des Animaux Aquatiques, BP 70, 29280 Plouzané, France
3 Centre Scientifique de Monaco, Avenue Saint-Martin, MC-98000 Monaco, Principality of Monaco



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  		Fig. 1. Protein concentrations in trout endolymph during the day:night cycle. Solid squares, proximal and open squares, distal endolymph. Values are means ± S.E.M. (N=4). Comparisons between reference (6:00 h) and day and night values were considered significant at P≤0.05.

 


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  		Fig. 2. Comparative analysis of endolymph samples collected during day and night periods. (A) Electrophoresis of proximal endolymph. Samples were run at 150 V for 1 h at 4°C on 12% SDS gels under reduced conditions. 5 µg protein was loaded per well. Proteins were stained with `silver stain plus kit'. (B) Analysis of the variations in endolymph composition was performed using N.I.H. software (see Materials and methods). Dark (day) and light (night) traces represent the level of band density on gel electrophoresis. Arrows represent the main variations observed in day and night samples. (C) Western blot of endolymph, showing the presence of specific precursors of otolith organic matrix in endolymph. Western blots were performed using 7.5% gel electrophoresis under the conditions described in Materials and methods. Nitrocellulose was probed using rabbit anti-OM (1/2000 dilution), followed by horse radish peroxidase-conjugated goat anti-rabbit globulin. (D) Densitometric analysis of the immunoblot in C. Dark (day) and light (night) traces represent the level of band density on western blotting. Arrows represent the main variations observed in day and night samples.

 


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  		Fig. 3. Daily variations of calcification factors in the endolymph. Schematic representation of overall variations in these factors observed during the day and night periods. OM, organic matrix; Sa, saturation state of aragonite.

 

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© The Company of Biologists Ltd 2003