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Immunohistochemical localization of two otolith matrix proteins in the otolith and inner ear of the rainbow trout, Oncorhynchus mykiss: comparative aspects between the adult inner ear and embryonic otocysts

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Abstract

The fish otolith consists mainly of calcium carbonate and organic matrices, the latter of which may play important roles in the process of otolith formation. We previously identified two otolith matrix proteins, named otolith matrix protein-1 (OMP-1) and otolin-1, from the rainbow trout, Oncorhynchus mykiss, and the chum salmon, O. keta. In this study, recombinant proteins corresponding to OMP-1 and otolin-1 were synthesized using yeast and bacterial expression systems, respectively, to produce specific antibodies against each protein. Immunohistochemical analysis using these antisera revealed that in the otoliths of adult fish, OMP-1 and otolin-1 were colocalized along the daily rings possibly formed by alternate deposition of calcium carbonate and organic matrices. In the adult inner ear, OMP-1 was produced at most of the saccular epithelium, while otolin-1 was produced at a limited part of cylindrical cells located at the marginal zone of the sensory epithelium. In the embryonic inner ear, these proteins had already existed in the otolith primordia when calcification had commenced. In addition, otolin-1 was localized in the fibrous materials connecting otolith primordia and sensory epithelium at this stage. These results indicate that these proteins are required as essential components for otolith formation and calcification.

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Acknowledgements

We are grateful to Dr. Goro Yoshizaki and Dr. Yutaka Takeuchi of the Department of Aquatic Biosciences, Tokyo University of Fisheries, for their generous gift of rainbow trout embryos and valuable discussion concerning embryonic development. We also thank Dr. M.N. Wilder of the Japan International Research Center for Agricultural Sciences for critical reading of the manuscript. This work was supported by Grants-in-Aid for Creative Basic Research (number 12NP0201) and for Scientific Research (numbers 12876025, 13660176, and 13660178) from the Ministry of Education, Culture, Sports, Science and Technology of Japan. The first author (E.M.) was supported by a Research Fellowship of the Japan Society for the Promotion of Science for Young Scientists.

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  1. Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo, 113-8657, Tokyo, Japan

    Hiromichi Nagasawa

  2. Laboratory of Bioorganic Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo, 113-8657, Tokyo, Japan

    Emi Murayama & Hiromichi Nagasawa

  3. Laboratory of Comparative Physiology, Division of Marine Bioscience, Graduate School of Fisheries Sciences, Hokkaido University, Minatomachi, Hakodate, 041-8611, Hokkaido, Japan

    Yasuaki Takagi

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  1. Emi Murayama
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  2. Yasuaki Takagi
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  3. Hiromichi Nagasawa
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Correspondence to Hiromichi Nagasawa.

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Murayama, E., Takagi, Y. & Nagasawa, H. Immunohistochemical localization of two otolith matrix proteins in the otolith and inner ear of the rainbow trout, Oncorhynchus mykiss: comparative aspects between the adult inner ear and embryonic otocysts. Histochem Cell Biol 121, 155–166 (2004). https://doi.org/10.1007/s00418-003-0605-5

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