Unique expression patterns of matrix metalloproteinases in regenerating newt limbs

Authors

  • Tomoko Kato,

    1. Department of Ophthalmology and Visual Science, Division of Frontier Medical Science, Biomedical Research, Graduate School of Biomedical Science, Hiroshima University, Hiroshima, Japan
    2. Developmental Biology Laboratory, Department of Biological Science, Graduate School of Science, Hiroshima University, Higashihiroshima, Japan
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    • Drs. Kato and Miyazaki contributed equally to this work.

  • Koyomi Miyazaki,

    1. Yoshizato MophoMatrix Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Higashihiroshima, Japan
    Current affiliation:
    1. Clock Cell Biology Group, Institute of Molecular and Cell Biology, National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan
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    • Drs. Kato and Miyazaki contributed equally to this work.

  • Keiko Shimizu-Nishikawa,

    1. Regenerative Biology Laboratory, Department of Biological Science, Graduate School of Science, Hiroshima University, Higashihiroshima, Japan
    2. Precursory Research for Embryonic Science and Technology, Japan Science and Technology Corporation, Higashihiroshima, Japan
    Current affiliation:
    1. Department of Biological Science, Faculty of Life and Environmental Science, Shimane University, Matsue, Japan
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  • Kazuko Koshiba,

    1. Yoshizato MophoMatrix Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Higashihiroshima, Japan
    Current affiliation:
    1. Department of Molecular Biology, Nara Institute of Science and Technology, Ikoma, Japan
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  • Masanobu Obara,

    1. Developmental Biology Laboratory, Department of Biological Science, Graduate School of Science, Hiroshima University, Higashihiroshima, Japan
    2. Tissue Regeneration Project, Hiroshima Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence, Japan Science and Technology Corporation, Hiroshima Prefectural Institute of Science and Technology, Higashihiroshima, Japan
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  • Hiromu K. Mishima,

    1. Department of Ophthalmology and Visual Science, Division of Frontier Medical Science, Biomedical Research, Graduate School of Biomedical Science, Hiroshima University, Hiroshima, Japan
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  • Katsutoshi Yoshizato

    Corresponding author
    1. Developmental Biology Laboratory, Department of Biological Science, Graduate School of Science, Hiroshima University, Higashihiroshima, Japan
    2. Yoshizato MophoMatrix Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Higashihiroshima, Japan
    3. Tissue Regeneration Project, Hiroshima Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence, Japan Science and Technology Corporation, Hiroshima Prefectural Institute of Science and Technology, Higashihiroshima, Japan
    • Developmental Biology Laboratory, Department of Biological Science, Graduate School of Science, Hiroshima University, Kagamiyama 1-3-1, Higashihiroshima, Hiroshima 739-8526, Japan
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Abstract

The process of regeneration of urodele limbs includes a drastic remodeling of extracellular matrices (ECMs) that is induced by matrix metalloproteinases (MMPs) and is thought to be one of the triggers of the regeneration. We studied this remodeling in limbs of Japanese newt, Cynops pyrrhogaster, by using five genes of newt MMPs (nMMPs) as probes: nMMP9, nMMP3/10-a, nMMP3/10-b, and nMMP13 that had been characterized previously, and nMMPe that was newly cloned in the present study. nMMPe was 502 amino acid residues long and showed a low homology to other known vertebrate MMPs. Reverse transcriptase-polymerase chain reactions analysis localized the transcript of nMMPe in the apical epidermal cap (AEC) and the non–blastemal wound epidermis but not in the blastemal mesenchyme or the normal epidermis. Northern blot analysis localized the transcripts of nMMP9, nMMP3/10-a, and nMMP13 in the bone of regenerating limbs, whereas those of nMMP3/10-b in AEC. mRNA in situ hybridization experiments identified the nMMP-expressing cells. nMMP9 gene was strongly expressed in chondrocytes of the cartilage of epiphysis. Of interest, basal cells of AEC, but not those of the normal skin, expressed nMMP3/10-b intensely. Immunohistochemical analysis showed that the nMMP9 proteins synthesized by chondrocytes were secreted and distributed widely in the basement membrane of bone and ECMs of the amputation plane. These nMMPs characterized in the present study might cooperatively work to remodel ECMs of regenerating limbs. Developmental Dynamics 226:366–376, 2003. © 2003 Wiley-Liss, Inc.

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