In the teleost fish otolith, the alternate deposition of CaCO 3 -rich and protein-rich layers results in the formation of daily increments. In order to clarify the mechanism of daily increment formation, a precise understanding of the relationship between ionic and organic controls of otolith growth is essential. In the present study, we studied diel variations in the aragonite saturation rate (S a ) of the endolymph and the mRNA expression of 2 major otolith matrix proteins, OMP-1 and otolin-1, in the saccular tissue. A new technique for simultaneously quantifying endolymph S a and mRNA expression in the saccular tissue from an individual rainbow trout was developed. Endolymph Ca 2+ levels, CO 2 partial pressure (P CO 2 ) and pH were simultaneously measured using an automatic pH/gas/electrolyte analyzer, and the S a was calculated. Total RNA was isolated from sacculi of individual fish after the endolymph was obtained. cDNAs were synthesized and used to quantify OMP-1 and otolin-1 mRNA expression levels by real-time PCR. Significant diel variations were observed in endolymph pH and P CO 2 levels in an antiphasic manner. Endolymph S a did not exhibit significant diel variations and was maintained at a value of more than 2.0, indicating that the endolymph was kept supersaturated with respect to aragonite during the day-night cycle. Expression of otolin-1 mRNA had apparent diel variations with high levels at night, whereas that of OMP-1 mRNA was almost constant. These data strongly suggest an organic control of daily increment formation in the otolith. The most probable candidate protein for daily increment formation is otolin-1.KEY WORDS: Calcification · mRNA expression · Endolymph · Daily increment · Aragonite · Otolith · Rainbow trout
Resale or republication not permitted without written consent of the publisherMar Ecol Prog Ser 294: [249][250][251][252][253][254][255][256] 2005 and the dark one is referred to as the D-zone (Kalish et al. 1995). Morphological examinations have revealed that the D-zones contain more organic matrices and less calcium carbonate relative to the L-zones (Mugiya & Muramatsu 1982, Watabe et al. 1982, Morales-Nin 1987; therefore, the alternate manner in which the deposition of mineral-rich and organic-matrix-rich layers occurs is manifested as daily increments. Indeed, Mugiya (1987) reported the discrete antiphasic diel cycles of 45 Ca and 3 H-glutamic-acid uptake into the otoliths in the rainbow trout in vitro and suggested that the daily increments of otoliths were formed by the antiphasic deposition of calcium and organic matrices. Daily variations in otolith calcification have also been demonstrated in the goldfish (Mugiya et al. 1981, Tohse & Mugiya 2002 and in the Atlantic salmon (Wright et al. 1992).The mechanism of these diel variations in otolith calcification and the deposition of the organic matrix is unknown. The otolith can be defined as acellular tissue that grows in an inner ear sacculus filled with endolymph. Consequently, endolymph must contain all the pr...