Expression of osteoblastic and osteoclastic genes during spontaneous regeneration and autotransplantation of goldfish scale: A new tool to study intramembranous bone regeneration

Thiparpa Aime Thamamongood, Ryo Furuya, Shunsuke Fukuba, Masahisa Nakamura, Nobuo Suzuki, Atsuhiko Hattori

    Research output: Contribution to journalArticle

    24 Citations (Scopus)

    Abstract

    Complementary DNA of osteoblast-specific genes (dlx5, runx2a, runx2b, osterix, RANKL, type I collagen, ALP, and osteocalcin) was cloned from goldfish (Carassius auratus) scale. Messenger RNA expressions were analyzed during spontaneous scale regeneration. Dlx5 had an early peak of expression on day 7, whereas osterix was constantly expressed during days 7-21. Runx2, a major osteoblastic transcription factor in mammalian bone, did not show any significant expression. The expressions of two functional genes, type I collagen and ALP, continually increased after day 7, while that of osteocalcin increased on day 14. As for osteoclastic markers, in addition to the cloning of two functional genes, TRAP and cathepsin K, in our previous study, we here cloned the transcription factor NFATc1 to use as an early osteoclastic marker. Using these bone markers, we investigate the signal key that controls the onset of scale resorption and regeneration by performing intra-scale-pocket autotransplantation of five groups of modified scales, namely, 1) methanol-fixed scale, 2) proteinase K-treated cell-free scale, 3) polarity reversal (upside-down) scale, 4) U-shape trimmed scale, and 5) circular-hole perforated scale. In this autotransplantation, each ontogenic scale was pulled out, modified, and then re-inserted into the same scale pocket. At post-transplant, inside the pockets of all modified transplant groups, new regenerating scales formed, attaching to the ongoing resorbed transplants. Autotransplantation of methanol-fixed scale, proteinase K-treated cell-free scale, and polarity reversal (upside-down) scale triggered scale resorption and scale regeneration. These two processes of scale resorption and regeneration occurred in accordance with osteoclastic and osteoblastic marker gene expressions. These results were microscopically confirmed using TRAP and ALP staining. Regarding the autotransplantation of U-shape trimmed and circular-hole perforated scales, new scales regenerated and grew at the trimmed/perforated part of each transplant, while scale resorption occurred apparently only around the trimmed/perforated area. In contrast, no scale resorption or regeneration was detected in sham transplantations. Our finding suggests that loss of correct cell-to-cell contact between the scale-pocket lining cells and the scale cortex cells is the key to switch on the onset of scale resorption and regeneration. Overall, the present study shows that goldfish scale regeneration shares similarities in gene expression with intramembranous bone regeneration. Improved understanding of goldfish scale regeneration will help elucidate the process of intramembranous bone regeneration and make goldfish scale a possible new tool to study bone regeneration.

    Original languageEnglish
    Pages (from-to)1240-1249
    Number of pages10
    JournalBone
    Volume50
    Issue number6
    DOIs
    Publication statusPublished - 2012 Jun

    Fingerprint

    Goldfish
    Bone Regeneration
    Autologous Transplantation
    Regeneration
    Genes
    Transplants
    Endopeptidase K
    Osteocalcin
    Collagen Type I
    Methanol
    Transcription Factors
    Cathepsin K
    Gene Expression
    Bone and Bones
    Osteoblasts
    Organism Cloning
    Complementary DNA
    Transplantation
    Staining and Labeling
    Messenger RNA

    Keywords

    • Cell-to-cell contact
    • Intramembranous bone regeneration
    • Osteoblastic genes
    • Osteoclastic genes
    • Scale autotransplantation
    • Teleost scale regeneration

    ASJC Scopus subject areas

    • Physiology
    • Endocrinology, Diabetes and Metabolism
    • Histology

    Cite this

    Expression of osteoblastic and osteoclastic genes during spontaneous regeneration and autotransplantation of goldfish scale : A new tool to study intramembranous bone regeneration. / Thamamongood, Thiparpa Aime; Furuya, Ryo; Fukuba, Shunsuke; Nakamura, Masahisa; Suzuki, Nobuo; Hattori, Atsuhiko.

    In: Bone, Vol. 50, No. 6, 06.2012, p. 1240-1249.

    Research output: Contribution to journalArticle

    Thamamongood, Thiparpa Aime ; Furuya, Ryo ; Fukuba, Shunsuke ; Nakamura, Masahisa ; Suzuki, Nobuo ; Hattori, Atsuhiko. / Expression of osteoblastic and osteoclastic genes during spontaneous regeneration and autotransplantation of goldfish scale : A new tool to study intramembranous bone regeneration. In: Bone. 2012 ; Vol. 50, No. 6. pp. 1240-1249.
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    abstract = "Complementary DNA of osteoblast-specific genes (dlx5, runx2a, runx2b, osterix, RANKL, type I collagen, ALP, and osteocalcin) was cloned from goldfish (Carassius auratus) scale. Messenger RNA expressions were analyzed during spontaneous scale regeneration. Dlx5 had an early peak of expression on day 7, whereas osterix was constantly expressed during days 7-21. Runx2, a major osteoblastic transcription factor in mammalian bone, did not show any significant expression. The expressions of two functional genes, type I collagen and ALP, continually increased after day 7, while that of osteocalcin increased on day 14. As for osteoclastic markers, in addition to the cloning of two functional genes, TRAP and cathepsin K, in our previous study, we here cloned the transcription factor NFATc1 to use as an early osteoclastic marker. Using these bone markers, we investigate the signal key that controls the onset of scale resorption and regeneration by performing intra-scale-pocket autotransplantation of five groups of modified scales, namely, 1) methanol-fixed scale, 2) proteinase K-treated cell-free scale, 3) polarity reversal (upside-down) scale, 4) U-shape trimmed scale, and 5) circular-hole perforated scale. In this autotransplantation, each ontogenic scale was pulled out, modified, and then re-inserted into the same scale pocket. At post-transplant, inside the pockets of all modified transplant groups, new regenerating scales formed, attaching to the ongoing resorbed transplants. Autotransplantation of methanol-fixed scale, proteinase K-treated cell-free scale, and polarity reversal (upside-down) scale triggered scale resorption and scale regeneration. These two processes of scale resorption and regeneration occurred in accordance with osteoclastic and osteoblastic marker gene expressions. These results were microscopically confirmed using TRAP and ALP staining. Regarding the autotransplantation of U-shape trimmed and circular-hole perforated scales, new scales regenerated and grew at the trimmed/perforated part of each transplant, while scale resorption occurred apparently only around the trimmed/perforated area. In contrast, no scale resorption or regeneration was detected in sham transplantations. Our finding suggests that loss of correct cell-to-cell contact between the scale-pocket lining cells and the scale cortex cells is the key to switch on the onset of scale resorption and regeneration. Overall, the present study shows that goldfish scale regeneration shares similarities in gene expression with intramembranous bone regeneration. Improved understanding of goldfish scale regeneration will help elucidate the process of intramembranous bone regeneration and make goldfish scale a possible new tool to study bone regeneration.",
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    AU - Thamamongood, Thiparpa Aime

    AU - Furuya, Ryo

    AU - Fukuba, Shunsuke

    AU - Nakamura, Masahisa

    AU - Suzuki, Nobuo

    AU - Hattori, Atsuhiko

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    N2 - Complementary DNA of osteoblast-specific genes (dlx5, runx2a, runx2b, osterix, RANKL, type I collagen, ALP, and osteocalcin) was cloned from goldfish (Carassius auratus) scale. Messenger RNA expressions were analyzed during spontaneous scale regeneration. Dlx5 had an early peak of expression on day 7, whereas osterix was constantly expressed during days 7-21. Runx2, a major osteoblastic transcription factor in mammalian bone, did not show any significant expression. The expressions of two functional genes, type I collagen and ALP, continually increased after day 7, while that of osteocalcin increased on day 14. As for osteoclastic markers, in addition to the cloning of two functional genes, TRAP and cathepsin K, in our previous study, we here cloned the transcription factor NFATc1 to use as an early osteoclastic marker. Using these bone markers, we investigate the signal key that controls the onset of scale resorption and regeneration by performing intra-scale-pocket autotransplantation of five groups of modified scales, namely, 1) methanol-fixed scale, 2) proteinase K-treated cell-free scale, 3) polarity reversal (upside-down) scale, 4) U-shape trimmed scale, and 5) circular-hole perforated scale. In this autotransplantation, each ontogenic scale was pulled out, modified, and then re-inserted into the same scale pocket. At post-transplant, inside the pockets of all modified transplant groups, new regenerating scales formed, attaching to the ongoing resorbed transplants. Autotransplantation of methanol-fixed scale, proteinase K-treated cell-free scale, and polarity reversal (upside-down) scale triggered scale resorption and scale regeneration. These two processes of scale resorption and regeneration occurred in accordance with osteoclastic and osteoblastic marker gene expressions. These results were microscopically confirmed using TRAP and ALP staining. Regarding the autotransplantation of U-shape trimmed and circular-hole perforated scales, new scales regenerated and grew at the trimmed/perforated part of each transplant, while scale resorption occurred apparently only around the trimmed/perforated area. In contrast, no scale resorption or regeneration was detected in sham transplantations. Our finding suggests that loss of correct cell-to-cell contact between the scale-pocket lining cells and the scale cortex cells is the key to switch on the onset of scale resorption and regeneration. Overall, the present study shows that goldfish scale regeneration shares similarities in gene expression with intramembranous bone regeneration. Improved understanding of goldfish scale regeneration will help elucidate the process of intramembranous bone regeneration and make goldfish scale a possible new tool to study bone regeneration.

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    KW - Teleost scale regeneration

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