The Experts below are selected from a list of 7104 Experts worldwide ranked by ideXlab platform

Hidetaka Sakai - One of the best experts on this subject based on the ideXlab platform.

  • Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) expression and possible function in mouse Tooth Germ development
    Journal of Molecular Histology, 2016
    Co-Authors: Kana Hasegawa, Hiroko Wada, Hidetaka Sakai, Kengo Nagata, Hiroaki Fujiwara, Naohisa Wada, Hirotaka Someya, Yurie Mikami, Tamotsu Kiyoshima
    Abstract:

    Abnormal expression of Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) is involved in the pathogenesis of FSHD. FRG1 is also important for the normal muscular and vascular development. Our previous study showed that FRG1 is one of the highly expressed genes in the mandible on embryonic day 10.5 (E10.5) than on E12.0. In this study, we investigated the temporospatial expression pattern of FRG1 mRNA and protein during the development of the mouse lower first molar, and also evaluated the subcellular localization of the FRG1 protein in mouse dental epithelial (mDE6) cells. The FRG1 expression was identified in the dental epithelial and mesenchymal cells at the initiation and bud stages. It was detected in the inner enamel epithelium at the cap and early bell stages. At the late bell and root formation stages, these signals were detected in ameloblasts and odontoblasts during the formation of enamel and dentin matrices, respectively. The FRG1 protein was localized in the cytoplasm in the mouse Tooth Germ in vivo, while FRG1 was detected predominantly in the nucleus and faintly in the cytoplasm in mDE6 cells in vitro. In mDE6 cells treated with bone morphogenetic protein 4 (BMP4), the protein expression of FRG1 increased in cytoplasm, suggesting that FRG1 may translocate to the cytoplasm. These findings suggest that FRG1 is involved in the morphogenesis of the Tooth Germ, as well as in the formation of enamel and dentin matrices and that FRG1 may play a role in the odontogenesis in the mouse following BMP4 stimulation.

  • The expression and function of thymosin beta 10 in Tooth Germ development.
    The International journal of developmental biology, 2013
    Co-Authors: Maho Shiotsuka, Tamotsu Kiyoshima, Hiroko Wada, Kana Hasegawa, Kengo Nagata, Hiroaki Fujiwara, Hirotaka Someya, Ichiro Takahashi, Makiko Kihara, Hidetaka Sakai
    Abstract:

    This study presents the expression pattern and functions of thymosin beta 10 (Tbeta10), a Tbeta4 homologue during the development of mouse lower first molars. An in situ signal of Tbeta10 was detected on embryonic day 10.5 (E10.5)-E15.5 mainly in dental mesenchymal cells as well as in dental epithelial cells, while Tbeta4 was expressed in dental epithelial cells. In the late bell stage, preodontoblasts with strong Tbeta10 expression and preameloblasts with strong Tbeta4 expression exhibited face-to-face localization, suggesting that an intimate cell-cell interaction might exist between preodontoblasts and preameloblasts to form dentin and enamel matrices. A strong Tbeta10 signal was found in odontoblasts in the lateral side of the dental pulp and in Hertwig’s epithelial root sheath, thus suggesting that Tbeta10 participates in the formation of the outline of the Tooth root. An inhibition assay using Tbeta10-siRNA in E11.0 mandibles showed significant growth inhibition in the Tooth Germ. The Tbeta10-siRNA-treated E15.0 Tooth Germ also showed significant developmental arrest. The number of Ki67-positive cells significantly decreased in the Tbeta10-siRNA-treated mandibles. The cellular proliferative activity was also significantly suppressed in Tb10-siRNA-treated cultured mouse dental pulpal and epithelial cells. These results indicate that developmental arrest of the Tooth Germ might be caused by a reduction in cell proliferative activity. The stage-specific temporal and spatial expression pattern of Tbeta10 in the developing Tooth Germ is indicative of multiple functions of Tbeta10 in the developmental course from initiation to root formation of the Tooth Germ.

  • Multiple functional involvement of Thymosin beta-4 in Tooth Germ development
    Histochemistry and cell biology, 2012
    Co-Authors: Yukiko Ookuma, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Haruyoshi Yamaza, Hiroko Wada, Kengo Nagata, Hiroaki Fujiwara, Kazuaki Nonaka, Hidetaka Sakai
    Abstract:

    Thymosin beta-4 (Tβ4) is known to be ubiquitously involved in the actin monomer sequestering on the cytoskeleton. Our previous study showed specific temporal and special in situ expression pattern of Tβ4 mRNA in dental epithelial and mesenchymal cells in the developing Tooth Germ of the mouse lower first molar. In this study, we examined the functional implications of Tβ4 in the developmental course of the mouse lower first molar. An inhibition assay using Tβ4 antisense sulfur-substituted oligodeoxynucleotide (AS S-ODN) in cultured embryonic day 11.0 (E11.0) mandibles showed a significant growth inhibition of the Tooth Germ. However, no growth arrest of the cultured E15.0 Tooth Germ was observed by using Tβ4 AS S-ODN. The Tβ4 knockdown led to significantly decreased expression levels of type II/III runt-related transcription factor 2 (Runx2) and nucleolin (Ncl) in the cultured E11.0 mandibles. Since our previous studies proved that the inhibition of type II/III Runx2 and Ncl translations resulted in the developmental arrest of the Tooth Germ in the cultured E11.0 mandible, Tβ4 appears to play roles in Tooth Germ development via the regulation of the type II/III Runx2 and Ncl expressions. Tβ4 knockdown also resulted in decreased secretion of matrix metalloproteinase (Mmp)-2, a reduced cell motility activity and upregulation of E-cadherin in dental epithelial mDE6 cells. These results suggest that Tβ4 plays multiple functional roles in odontogenic epithelial cells in the early stages of Tooth Germ development by regulating the expression of odontogenesis-related genes.

  • in situ expression of the mitochondrial atpase6 gene in the developing Tooth Germ of the mouse lower first molar
    Journal of Molecular Histology, 2011
    Co-Authors: Jun-ya Honda, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Hiroko Wada, Kengo Nagata, Hiroaki Fujiwara, Yukiko Ookuma, Ichiro Takahashi, Maho Shiotsuka, Hidetaka Sakai
    Abstract:

    We previously performed cDNA subtraction between the mouse mandibles on embryonic day 10.5 (E10.5) in the pre-initiation stage of the odontogenesis and E12.0 in the late initiation stage to identify genes expressed at its beginning. Adenosine triphosphate synthase subunit a (Atpase6) is one of the highly expressed genes in the E12.0 mandible including Tooth Germs. In situ hybridization was conducted using the mouse mandibular first molar from E10.5 to E18.0 to determine the precise expression patterns of Atpase6 mRNA in the developing Tooth Germ. Atpase6 mRNA was strongly expressed in the presumptive dental epithelium and the underlying mesenchyme at E10.5, and in the thickened dental epithelium at E12.0 and E13.0. Strong in situ signals were observed in the epithelium at E14.0, and in the enamel organ excluded the area of the primary enamel knot at E15.0. Atpase6 was strongly expressed in the inner enamel epithelium, the adjacent stratum intermedium, and the outer enamel epithelium in the cervical loops from E16.0 to E18.0. In addition, strong Atpase6 signals were coincidently demonstrated in various developing cranio-facial organs. These results suggest that Atpase6 participates in the high energy-utilizing functions of the cells related to the initiation and the development of the Tooth Germ as well as those of the other cranio-facial organs.

  • glycolytic enzyme pgk1 is strongly expressed in the developing Tooth Germ of the mouse lower first molar
    Histology and Histopathology, 2008
    Co-Authors: Jun-ya Honda, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Haruyoshi Yamaza, Kengo Nagata, Ming Xie, Keiko Takahashi, Norio Enoki, Akihiko Nakashima, Hidetaka Sakai
    Abstract:

    This study examined detailed in situ expression patterns and possible functional roles of phosphoglycerate kinase 1 (Pgk1) gene in the developing Tooth Germ of the mouse lower first molar. The strong expression of Pgk1 mRNA was seen in the odontogenic epithelial cells and surrounding mesenchymal cells of the Tooth Germ from embryonic day 10.5 (E10.5) to E18.0. Western blotting analysis demonstrated that Pgk1 protein formed 84-kDa protein complex in these embryonic organs. The results of immunoprecipitation-western blotting also suggested this complex to be formed with glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Moreover, the immunofluorescence expression of those proteins was shown to overlap each other in the Tooth Germ at E15.0. A strong immunofluorescence expression of both Pgk1 and GAPDH also corresponded to the in situ expression of those mRNAs. These results suggested that Pgk1 plays some functional roles in the development of Tooth Germ and other embryonic organs by forming protein complex with GAPDH.

Tamotsu Kiyoshima - One of the best experts on this subject based on the ideXlab platform.

  • wnt β catenin signaling which is activated in odontomas reduces sema3a expression to regulate odontogenic epithelial cell proliferation and Tooth Germ development
    Scientific Reports, 2019
    Co-Authors: Shinsuke Fujii, Tamotsu Kiyoshima, Kengo Nagata, Shinji Matsumoto, Ken-ichi Kohashi, Akira Kikuchi, Yoshinao Oda, Naohisa Wada
    Abstract:

    Odontomas, developmental anomalies of Tooth Germ, frequently occur in familial adenomatous polyposis patients with activated Wnt/β-catenin signaling. However, roles of Wnt/β-catenin signaling in odontomas or odontogenic cells are unclear. Herein, we investigated β-catenin expression in odontomas and functions of Wnt/β-catenin signaling in Tooth Germ development. β-catenin frequently accumulated in nucleus and/or cellular cytoplasm of odontogenic epithelial cells in human odontoma specimens, immunohistochemically. Wnt/β-catenin signaling inhibited odontogenic epithelial cell proliferation in both cell line and Tooth Germ development, while inducing immature epithelial bud formation. We identified Semaphorin 3A (Sema3A) as a downstream molecule of Wnt/β-catenin signaling and showed that Wnt/β-catenin signaling-dependent reduction of Sema3A expression resulted in suppressed odontogenic epithelial cell proliferation. Sema3A expression is required in appropriate epithelial budding morphogenesis. These results suggest that Wnt/β-catenin signaling negatively regulates odontogenic epithelial cell proliferation and Tooth Germ development through decreased-Sema3A expression, and aberrant activation of Wnt/β-catenin signaling may associate with odontoma formation.

  • Wnt/β-catenin signaling, which is activated in odontomas, reduces Sema3A expression to regulate odontogenic epithelial cell proliferation and Tooth Germ development
    Nature Publishing Group, 2019
    Co-Authors: Shinsuke Fujii, Tamotsu Kiyoshima, Kengo Nagata, Shinji Matsumoto, Ken-ichi Kohashi, Akira Kikuchi, Yoshinao Oda, Naohisa Wada
    Abstract:

    Abstract Odontomas, developmental anomalies of Tooth Germ, frequently occur in familial adenomatous polyposis patients with activated Wnt/β-catenin signaling. However, roles of Wnt/β-catenin signaling in odontomas or odontogenic cells are unclear. Herein, we investigated β-catenin expression in odontomas and functions of Wnt/β-catenin signaling in Tooth Germ development. β-catenin frequently accumulated in nucleus and/or cellular cytoplasm of odontogenic epithelial cells in human odontoma specimens, immunohistochemically. Wnt/β-catenin signaling inhibited odontogenic epithelial cell proliferation in both cell line and Tooth Germ development, while inducing immature epithelial bud formation. We identified Semaphorin 3A (Sema3A) as a downstream molecule of Wnt/β-catenin signaling and showed that Wnt/β-catenin signaling-dependent reduction of Sema3A expression resulted in suppressed odontogenic epithelial cell proliferation. Sema3A expression is required in appropriate epithelial budding morphogenesis. These results suggest that Wnt/β-catenin signaling negatively regulates odontogenic epithelial cell proliferation and Tooth Germ development through decreased-Sema3A expression, and aberrant activation of Wnt/β-catenin signaling may associate with odontoma formation

  • Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) expression and possible function in mouse Tooth Germ development
    Journal of Molecular Histology, 2016
    Co-Authors: Kana Hasegawa, Hiroko Wada, Hidetaka Sakai, Kengo Nagata, Hiroaki Fujiwara, Naohisa Wada, Hirotaka Someya, Yurie Mikami, Tamotsu Kiyoshima
    Abstract:

    Abnormal expression of Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) is involved in the pathogenesis of FSHD. FRG1 is also important for the normal muscular and vascular development. Our previous study showed that FRG1 is one of the highly expressed genes in the mandible on embryonic day 10.5 (E10.5) than on E12.0. In this study, we investigated the temporospatial expression pattern of FRG1 mRNA and protein during the development of the mouse lower first molar, and also evaluated the subcellular localization of the FRG1 protein in mouse dental epithelial (mDE6) cells. The FRG1 expression was identified in the dental epithelial and mesenchymal cells at the initiation and bud stages. It was detected in the inner enamel epithelium at the cap and early bell stages. At the late bell and root formation stages, these signals were detected in ameloblasts and odontoblasts during the formation of enamel and dentin matrices, respectively. The FRG1 protein was localized in the cytoplasm in the mouse Tooth Germ in vivo, while FRG1 was detected predominantly in the nucleus and faintly in the cytoplasm in mDE6 cells in vitro. In mDE6 cells treated with bone morphogenetic protein 4 (BMP4), the protein expression of FRG1 increased in cytoplasm, suggesting that FRG1 may translocate to the cytoplasm. These findings suggest that FRG1 is involved in the morphogenesis of the Tooth Germ, as well as in the formation of enamel and dentin matrices and that FRG1 may play a role in the odontogenesis in the mouse following BMP4 stimulation.

  • The expression and function of thymosin beta 10 in Tooth Germ development.
    The International journal of developmental biology, 2013
    Co-Authors: Maho Shiotsuka, Tamotsu Kiyoshima, Hiroko Wada, Kana Hasegawa, Kengo Nagata, Hiroaki Fujiwara, Hirotaka Someya, Ichiro Takahashi, Makiko Kihara, Hidetaka Sakai
    Abstract:

    This study presents the expression pattern and functions of thymosin beta 10 (Tbeta10), a Tbeta4 homologue during the development of mouse lower first molars. An in situ signal of Tbeta10 was detected on embryonic day 10.5 (E10.5)-E15.5 mainly in dental mesenchymal cells as well as in dental epithelial cells, while Tbeta4 was expressed in dental epithelial cells. In the late bell stage, preodontoblasts with strong Tbeta10 expression and preameloblasts with strong Tbeta4 expression exhibited face-to-face localization, suggesting that an intimate cell-cell interaction might exist between preodontoblasts and preameloblasts to form dentin and enamel matrices. A strong Tbeta10 signal was found in odontoblasts in the lateral side of the dental pulp and in Hertwig’s epithelial root sheath, thus suggesting that Tbeta10 participates in the formation of the outline of the Tooth root. An inhibition assay using Tbeta10-siRNA in E11.0 mandibles showed significant growth inhibition in the Tooth Germ. The Tbeta10-siRNA-treated E15.0 Tooth Germ also showed significant developmental arrest. The number of Ki67-positive cells significantly decreased in the Tbeta10-siRNA-treated mandibles. The cellular proliferative activity was also significantly suppressed in Tb10-siRNA-treated cultured mouse dental pulpal and epithelial cells. These results indicate that developmental arrest of the Tooth Germ might be caused by a reduction in cell proliferative activity. The stage-specific temporal and spatial expression pattern of Tbeta10 in the developing Tooth Germ is indicative of multiple functions of Tbeta10 in the developmental course from initiation to root formation of the Tooth Germ.

  • Multiple functional involvement of Thymosin beta-4 in Tooth Germ development
    Histochemistry and cell biology, 2012
    Co-Authors: Yukiko Ookuma, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Haruyoshi Yamaza, Hiroko Wada, Kengo Nagata, Hiroaki Fujiwara, Kazuaki Nonaka, Hidetaka Sakai
    Abstract:

    Thymosin beta-4 (Tβ4) is known to be ubiquitously involved in the actin monomer sequestering on the cytoskeleton. Our previous study showed specific temporal and special in situ expression pattern of Tβ4 mRNA in dental epithelial and mesenchymal cells in the developing Tooth Germ of the mouse lower first molar. In this study, we examined the functional implications of Tβ4 in the developmental course of the mouse lower first molar. An inhibition assay using Tβ4 antisense sulfur-substituted oligodeoxynucleotide (AS S-ODN) in cultured embryonic day 11.0 (E11.0) mandibles showed a significant growth inhibition of the Tooth Germ. However, no growth arrest of the cultured E15.0 Tooth Germ was observed by using Tβ4 AS S-ODN. The Tβ4 knockdown led to significantly decreased expression levels of type II/III runt-related transcription factor 2 (Runx2) and nucleolin (Ncl) in the cultured E11.0 mandibles. Since our previous studies proved that the inhibition of type II/III Runx2 and Ncl translations resulted in the developmental arrest of the Tooth Germ in the cultured E11.0 mandible, Tβ4 appears to play roles in Tooth Germ development via the regulation of the type II/III Runx2 and Ncl expressions. Tβ4 knockdown also resulted in decreased secretion of matrix metalloproteinase (Mmp)-2, a reduced cell motility activity and upregulation of E-cadherin in dental epithelial mDE6 cells. These results suggest that Tβ4 plays multiple functional roles in odontogenic epithelial cells in the early stages of Tooth Germ development by regulating the expression of odontogenesis-related genes.

Ieyoshi Kobayashi - One of the best experts on this subject based on the ideXlab platform.

  • Multiple functional involvement of Thymosin beta-4 in Tooth Germ development
    Histochemistry and cell biology, 2012
    Co-Authors: Yukiko Ookuma, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Haruyoshi Yamaza, Hiroko Wada, Kengo Nagata, Hiroaki Fujiwara, Kazuaki Nonaka, Hidetaka Sakai
    Abstract:

    Thymosin beta-4 (Tβ4) is known to be ubiquitously involved in the actin monomer sequestering on the cytoskeleton. Our previous study showed specific temporal and special in situ expression pattern of Tβ4 mRNA in dental epithelial and mesenchymal cells in the developing Tooth Germ of the mouse lower first molar. In this study, we examined the functional implications of Tβ4 in the developmental course of the mouse lower first molar. An inhibition assay using Tβ4 antisense sulfur-substituted oligodeoxynucleotide (AS S-ODN) in cultured embryonic day 11.0 (E11.0) mandibles showed a significant growth inhibition of the Tooth Germ. However, no growth arrest of the cultured E15.0 Tooth Germ was observed by using Tβ4 AS S-ODN. The Tβ4 knockdown led to significantly decreased expression levels of type II/III runt-related transcription factor 2 (Runx2) and nucleolin (Ncl) in the cultured E11.0 mandibles. Since our previous studies proved that the inhibition of type II/III Runx2 and Ncl translations resulted in the developmental arrest of the Tooth Germ in the cultured E11.0 mandible, Tβ4 appears to play roles in Tooth Germ development via the regulation of the type II/III Runx2 and Ncl expressions. Tβ4 knockdown also resulted in decreased secretion of matrix metalloproteinase (Mmp)-2, a reduced cell motility activity and upregulation of E-cadherin in dental epithelial mDE6 cells. These results suggest that Tβ4 plays multiple functional roles in odontogenic epithelial cells in the early stages of Tooth Germ development by regulating the expression of odontogenesis-related genes.

  • in situ expression of the mitochondrial atpase6 gene in the developing Tooth Germ of the mouse lower first molar
    Journal of Molecular Histology, 2011
    Co-Authors: Jun-ya Honda, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Hiroko Wada, Kengo Nagata, Hiroaki Fujiwara, Yukiko Ookuma, Ichiro Takahashi, Maho Shiotsuka, Hidetaka Sakai
    Abstract:

    We previously performed cDNA subtraction between the mouse mandibles on embryonic day 10.5 (E10.5) in the pre-initiation stage of the odontogenesis and E12.0 in the late initiation stage to identify genes expressed at its beginning. Adenosine triphosphate synthase subunit a (Atpase6) is one of the highly expressed genes in the E12.0 mandible including Tooth Germs. In situ hybridization was conducted using the mouse mandibular first molar from E10.5 to E18.0 to determine the precise expression patterns of Atpase6 mRNA in the developing Tooth Germ. Atpase6 mRNA was strongly expressed in the presumptive dental epithelium and the underlying mesenchyme at E10.5, and in the thickened dental epithelium at E12.0 and E13.0. Strong in situ signals were observed in the epithelium at E14.0, and in the enamel organ excluded the area of the primary enamel knot at E15.0. Atpase6 was strongly expressed in the inner enamel epithelium, the adjacent stratum intermedium, and the outer enamel epithelium in the cervical loops from E16.0 to E18.0. In addition, strong Atpase6 signals were coincidently demonstrated in various developing cranio-facial organs. These results suggest that Atpase6 participates in the high energy-utilizing functions of the cells related to the initiation and the development of the Tooth Germ as well as those of the other cranio-facial organs.

  • glycolytic enzyme pgk1 is strongly expressed in the developing Tooth Germ of the mouse lower first molar
    Histology and Histopathology, 2008
    Co-Authors: Jun-ya Honda, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Haruyoshi Yamaza, Kengo Nagata, Ming Xie, Keiko Takahashi, Norio Enoki, Akihiko Nakashima, Hidetaka Sakai
    Abstract:

    This study examined detailed in situ expression patterns and possible functional roles of phosphoglycerate kinase 1 (Pgk1) gene in the developing Tooth Germ of the mouse lower first molar. The strong expression of Pgk1 mRNA was seen in the odontogenic epithelial cells and surrounding mesenchymal cells of the Tooth Germ from embryonic day 10.5 (E10.5) to E18.0. Western blotting analysis demonstrated that Pgk1 protein formed 84-kDa protein complex in these embryonic organs. The results of immunoprecipitation-western blotting also suggested this complex to be formed with glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Moreover, the immunofluorescence expression of those proteins was shown to overlap each other in the Tooth Germ at E15.0. A strong immunofluorescence expression of both Pgk1 and GAPDH also corresponded to the in situ expression of those mRNAs. These results suggested that Pgk1 plays some functional roles in the development of Tooth Germ and other embryonic organs by forming protein complex with GAPDH.

  • type ii iii runx2 cbfa1 is required for Tooth Germ development
    Bone, 2006
    Co-Authors: Ieyoshi Kobayashi, Tamotsu Kiyoshima, Kou Matsuo, Jun-ya Honda, Hiroko Wada, Kazuaki Nonaka, Kiyoshi Koyano, Hidetaka Sakai
    Abstract:

    Runx2/Cbfa1 is an essential transcription factor for osteoblast differentiation and bone formation. Runx2/Cbfa1 knockout mice showed both a complete lack of ossification and the developmental arrest of Tooth Germ. We here report Runx2/Cbfa1 isoform-type specific functional roles in the development of Tooth Germ by the administration of antisense phosphorothioate oligodioxynucleotides (S-ODNs) into cultured mouse mandibles. The administration of type II/III Runx2/Cbfa1 antisense S-ODNs into the culture media resulted in an arrest of Tooth Germ growth at the bud-like stage in cultured mandible taken from the 11-day-old embryos, while also causing the inhibition of the differentiation of odontogenic cells into ameloblast and odontoblast in cultured Tooth Germs taken from the 15-day-old embryos. The expression of dentin matrix protein 1, dentin sialophosphoprotein, amelogenin, and ameloblastin was shown to be markedly suppressed in cultured Tooth Germ by the semi-quantitative RT-PCR. Meanwhile, no developmental arrest of Tooth Germ, no inhibition of gene expression, or differentiation of odontogenic cells was observed in samples treated with the type I Runx2/Cbfa1 antisense S-ODNs. The same findings were also observed in either the control or the sense and random sequence S-ODNs-treated samples. These data indicate that the type II/III Runx2/Cbfa1 isoform is closely related to the development and differentiation of Tooth Germ.

  • Type II/III Runx2/Cbfa1 is required for Tooth Germ development
    Bone, 2005
    Co-Authors: Ieyoshi Kobayashi, Tamotsu Kiyoshima, Kou Matsuo, Jun-ya Honda, Hiroko Wada, Kazuaki Nonaka, Kiyoshi Koyano, Hidetaka Sakai
    Abstract:

    Runx2/Cbfa1 is an essential transcription factor for osteoblast differentiation and bone formation. Runx2/Cbfa1 knockout mice showed both a complete lack of ossification and the developmental arrest of Tooth Germ. We here report Runx2/Cbfa1 isoform-type specific functional roles in the development of Tooth Germ by the administration of antisense phosphorothioate oligodioxynucleotides (S-ODNs) into cultured mouse mandibles. The administration of type II/III Runx2/Cbfa1 antisense S-ODNs into the culture media resulted in an arrest of Tooth Germ growth at the bud-like stage in cultured mandible taken from the 11-day-old embryos, while also causing the inhibition of the differentiation of odontogenic cells into ameloblast and odontoblast in cultured Tooth Germs taken from the 15-day-old embryos. The expression of dentin matrix protein 1, dentin sialophosphoprotein, amelogenin, and ameloblastin was shown to be markedly suppressed in cultured Tooth Germ by the semi-quantitative RT-PCR. Meanwhile, no developmental arrest of Tooth Germ, no inhibition of gene expression, or differentiation of odontogenic cells was observed in samples treated with the type I Runx2/Cbfa1 antisense S-ODNs. The same findings were also observed in either the control or the sense and random sequence S-ODNs-treated samples. These data indicate that the type II/III Runx2/Cbfa1 isoform is closely related to the development and differentiation of Tooth Germ.

Hiroko Wada - One of the best experts on this subject based on the ideXlab platform.

  • Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) expression and possible function in mouse Tooth Germ development
    Journal of Molecular Histology, 2016
    Co-Authors: Kana Hasegawa, Hiroko Wada, Hidetaka Sakai, Kengo Nagata, Hiroaki Fujiwara, Naohisa Wada, Hirotaka Someya, Yurie Mikami, Tamotsu Kiyoshima
    Abstract:

    Abnormal expression of Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) is involved in the pathogenesis of FSHD. FRG1 is also important for the normal muscular and vascular development. Our previous study showed that FRG1 is one of the highly expressed genes in the mandible on embryonic day 10.5 (E10.5) than on E12.0. In this study, we investigated the temporospatial expression pattern of FRG1 mRNA and protein during the development of the mouse lower first molar, and also evaluated the subcellular localization of the FRG1 protein in mouse dental epithelial (mDE6) cells. The FRG1 expression was identified in the dental epithelial and mesenchymal cells at the initiation and bud stages. It was detected in the inner enamel epithelium at the cap and early bell stages. At the late bell and root formation stages, these signals were detected in ameloblasts and odontoblasts during the formation of enamel and dentin matrices, respectively. The FRG1 protein was localized in the cytoplasm in the mouse Tooth Germ in vivo, while FRG1 was detected predominantly in the nucleus and faintly in the cytoplasm in mDE6 cells in vitro. In mDE6 cells treated with bone morphogenetic protein 4 (BMP4), the protein expression of FRG1 increased in cytoplasm, suggesting that FRG1 may translocate to the cytoplasm. These findings suggest that FRG1 is involved in the morphogenesis of the Tooth Germ, as well as in the formation of enamel and dentin matrices and that FRG1 may play a role in the odontogenesis in the mouse following BMP4 stimulation.

  • The expression and function of thymosin beta 10 in Tooth Germ development.
    The International journal of developmental biology, 2013
    Co-Authors: Maho Shiotsuka, Tamotsu Kiyoshima, Hiroko Wada, Kana Hasegawa, Kengo Nagata, Hiroaki Fujiwara, Hirotaka Someya, Ichiro Takahashi, Makiko Kihara, Hidetaka Sakai
    Abstract:

    This study presents the expression pattern and functions of thymosin beta 10 (Tbeta10), a Tbeta4 homologue during the development of mouse lower first molars. An in situ signal of Tbeta10 was detected on embryonic day 10.5 (E10.5)-E15.5 mainly in dental mesenchymal cells as well as in dental epithelial cells, while Tbeta4 was expressed in dental epithelial cells. In the late bell stage, preodontoblasts with strong Tbeta10 expression and preameloblasts with strong Tbeta4 expression exhibited face-to-face localization, suggesting that an intimate cell-cell interaction might exist between preodontoblasts and preameloblasts to form dentin and enamel matrices. A strong Tbeta10 signal was found in odontoblasts in the lateral side of the dental pulp and in Hertwig’s epithelial root sheath, thus suggesting that Tbeta10 participates in the formation of the outline of the Tooth root. An inhibition assay using Tbeta10-siRNA in E11.0 mandibles showed significant growth inhibition in the Tooth Germ. The Tbeta10-siRNA-treated E15.0 Tooth Germ also showed significant developmental arrest. The number of Ki67-positive cells significantly decreased in the Tbeta10-siRNA-treated mandibles. The cellular proliferative activity was also significantly suppressed in Tb10-siRNA-treated cultured mouse dental pulpal and epithelial cells. These results indicate that developmental arrest of the Tooth Germ might be caused by a reduction in cell proliferative activity. The stage-specific temporal and spatial expression pattern of Tbeta10 in the developing Tooth Germ is indicative of multiple functions of Tbeta10 in the developmental course from initiation to root formation of the Tooth Germ.

  • Multiple functional involvement of Thymosin beta-4 in Tooth Germ development
    Histochemistry and cell biology, 2012
    Co-Authors: Yukiko Ookuma, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Haruyoshi Yamaza, Hiroko Wada, Kengo Nagata, Hiroaki Fujiwara, Kazuaki Nonaka, Hidetaka Sakai
    Abstract:

    Thymosin beta-4 (Tβ4) is known to be ubiquitously involved in the actin monomer sequestering on the cytoskeleton. Our previous study showed specific temporal and special in situ expression pattern of Tβ4 mRNA in dental epithelial and mesenchymal cells in the developing Tooth Germ of the mouse lower first molar. In this study, we examined the functional implications of Tβ4 in the developmental course of the mouse lower first molar. An inhibition assay using Tβ4 antisense sulfur-substituted oligodeoxynucleotide (AS S-ODN) in cultured embryonic day 11.0 (E11.0) mandibles showed a significant growth inhibition of the Tooth Germ. However, no growth arrest of the cultured E15.0 Tooth Germ was observed by using Tβ4 AS S-ODN. The Tβ4 knockdown led to significantly decreased expression levels of type II/III runt-related transcription factor 2 (Runx2) and nucleolin (Ncl) in the cultured E11.0 mandibles. Since our previous studies proved that the inhibition of type II/III Runx2 and Ncl translations resulted in the developmental arrest of the Tooth Germ in the cultured E11.0 mandible, Tβ4 appears to play roles in Tooth Germ development via the regulation of the type II/III Runx2 and Ncl expressions. Tβ4 knockdown also resulted in decreased secretion of matrix metalloproteinase (Mmp)-2, a reduced cell motility activity and upregulation of E-cadherin in dental epithelial mDE6 cells. These results suggest that Tβ4 plays multiple functional roles in odontogenic epithelial cells in the early stages of Tooth Germ development by regulating the expression of odontogenesis-related genes.

  • in situ expression of the mitochondrial atpase6 gene in the developing Tooth Germ of the mouse lower first molar
    Journal of Molecular Histology, 2011
    Co-Authors: Jun-ya Honda, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Hiroko Wada, Kengo Nagata, Hiroaki Fujiwara, Yukiko Ookuma, Ichiro Takahashi, Maho Shiotsuka, Hidetaka Sakai
    Abstract:

    We previously performed cDNA subtraction between the mouse mandibles on embryonic day 10.5 (E10.5) in the pre-initiation stage of the odontogenesis and E12.0 in the late initiation stage to identify genes expressed at its beginning. Adenosine triphosphate synthase subunit a (Atpase6) is one of the highly expressed genes in the E12.0 mandible including Tooth Germs. In situ hybridization was conducted using the mouse mandibular first molar from E10.5 to E18.0 to determine the precise expression patterns of Atpase6 mRNA in the developing Tooth Germ. Atpase6 mRNA was strongly expressed in the presumptive dental epithelium and the underlying mesenchyme at E10.5, and in the thickened dental epithelium at E12.0 and E13.0. Strong in situ signals were observed in the epithelium at E14.0, and in the enamel organ excluded the area of the primary enamel knot at E15.0. Atpase6 was strongly expressed in the inner enamel epithelium, the adjacent stratum intermedium, and the outer enamel epithelium in the cervical loops from E16.0 to E18.0. In addition, strong Atpase6 signals were coincidently demonstrated in various developing cranio-facial organs. These results suggest that Atpase6 participates in the high energy-utilizing functions of the cells related to the initiation and the development of the Tooth Germ as well as those of the other cranio-facial organs.

  • type ii iii runx2 cbfa1 is required for Tooth Germ development
    Bone, 2006
    Co-Authors: Ieyoshi Kobayashi, Tamotsu Kiyoshima, Kou Matsuo, Jun-ya Honda, Hiroko Wada, Kazuaki Nonaka, Kiyoshi Koyano, Hidetaka Sakai
    Abstract:

    Runx2/Cbfa1 is an essential transcription factor for osteoblast differentiation and bone formation. Runx2/Cbfa1 knockout mice showed both a complete lack of ossification and the developmental arrest of Tooth Germ. We here report Runx2/Cbfa1 isoform-type specific functional roles in the development of Tooth Germ by the administration of antisense phosphorothioate oligodioxynucleotides (S-ODNs) into cultured mouse mandibles. The administration of type II/III Runx2/Cbfa1 antisense S-ODNs into the culture media resulted in an arrest of Tooth Germ growth at the bud-like stage in cultured mandible taken from the 11-day-old embryos, while also causing the inhibition of the differentiation of odontogenic cells into ameloblast and odontoblast in cultured Tooth Germs taken from the 15-day-old embryos. The expression of dentin matrix protein 1, dentin sialophosphoprotein, amelogenin, and ameloblastin was shown to be markedly suppressed in cultured Tooth Germ by the semi-quantitative RT-PCR. Meanwhile, no developmental arrest of Tooth Germ, no inhibition of gene expression, or differentiation of odontogenic cells was observed in samples treated with the type I Runx2/Cbfa1 antisense S-ODNs. The same findings were also observed in either the control or the sense and random sequence S-ODNs-treated samples. These data indicate that the type II/III Runx2/Cbfa1 isoform is closely related to the development and differentiation of Tooth Germ.

Kengo Nagata - One of the best experts on this subject based on the ideXlab platform.

  • wnt β catenin signaling which is activated in odontomas reduces sema3a expression to regulate odontogenic epithelial cell proliferation and Tooth Germ development
    Scientific Reports, 2019
    Co-Authors: Shinsuke Fujii, Tamotsu Kiyoshima, Kengo Nagata, Shinji Matsumoto, Ken-ichi Kohashi, Akira Kikuchi, Yoshinao Oda, Naohisa Wada
    Abstract:

    Odontomas, developmental anomalies of Tooth Germ, frequently occur in familial adenomatous polyposis patients with activated Wnt/β-catenin signaling. However, roles of Wnt/β-catenin signaling in odontomas or odontogenic cells are unclear. Herein, we investigated β-catenin expression in odontomas and functions of Wnt/β-catenin signaling in Tooth Germ development. β-catenin frequently accumulated in nucleus and/or cellular cytoplasm of odontogenic epithelial cells in human odontoma specimens, immunohistochemically. Wnt/β-catenin signaling inhibited odontogenic epithelial cell proliferation in both cell line and Tooth Germ development, while inducing immature epithelial bud formation. We identified Semaphorin 3A (Sema3A) as a downstream molecule of Wnt/β-catenin signaling and showed that Wnt/β-catenin signaling-dependent reduction of Sema3A expression resulted in suppressed odontogenic epithelial cell proliferation. Sema3A expression is required in appropriate epithelial budding morphogenesis. These results suggest that Wnt/β-catenin signaling negatively regulates odontogenic epithelial cell proliferation and Tooth Germ development through decreased-Sema3A expression, and aberrant activation of Wnt/β-catenin signaling may associate with odontoma formation.

  • Wnt/β-catenin signaling, which is activated in odontomas, reduces Sema3A expression to regulate odontogenic epithelial cell proliferation and Tooth Germ development
    Nature Publishing Group, 2019
    Co-Authors: Shinsuke Fujii, Tamotsu Kiyoshima, Kengo Nagata, Shinji Matsumoto, Ken-ichi Kohashi, Akira Kikuchi, Yoshinao Oda, Naohisa Wada
    Abstract:

    Abstract Odontomas, developmental anomalies of Tooth Germ, frequently occur in familial adenomatous polyposis patients with activated Wnt/β-catenin signaling. However, roles of Wnt/β-catenin signaling in odontomas or odontogenic cells are unclear. Herein, we investigated β-catenin expression in odontomas and functions of Wnt/β-catenin signaling in Tooth Germ development. β-catenin frequently accumulated in nucleus and/or cellular cytoplasm of odontogenic epithelial cells in human odontoma specimens, immunohistochemically. Wnt/β-catenin signaling inhibited odontogenic epithelial cell proliferation in both cell line and Tooth Germ development, while inducing immature epithelial bud formation. We identified Semaphorin 3A (Sema3A) as a downstream molecule of Wnt/β-catenin signaling and showed that Wnt/β-catenin signaling-dependent reduction of Sema3A expression resulted in suppressed odontogenic epithelial cell proliferation. Sema3A expression is required in appropriate epithelial budding morphogenesis. These results suggest that Wnt/β-catenin signaling negatively regulates odontogenic epithelial cell proliferation and Tooth Germ development through decreased-Sema3A expression, and aberrant activation of Wnt/β-catenin signaling may associate with odontoma formation

  • Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) expression and possible function in mouse Tooth Germ development
    Journal of Molecular Histology, 2016
    Co-Authors: Kana Hasegawa, Hiroko Wada, Hidetaka Sakai, Kengo Nagata, Hiroaki Fujiwara, Naohisa Wada, Hirotaka Someya, Yurie Mikami, Tamotsu Kiyoshima
    Abstract:

    Abnormal expression of Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) is involved in the pathogenesis of FSHD. FRG1 is also important for the normal muscular and vascular development. Our previous study showed that FRG1 is one of the highly expressed genes in the mandible on embryonic day 10.5 (E10.5) than on E12.0. In this study, we investigated the temporospatial expression pattern of FRG1 mRNA and protein during the development of the mouse lower first molar, and also evaluated the subcellular localization of the FRG1 protein in mouse dental epithelial (mDE6) cells. The FRG1 expression was identified in the dental epithelial and mesenchymal cells at the initiation and bud stages. It was detected in the inner enamel epithelium at the cap and early bell stages. At the late bell and root formation stages, these signals were detected in ameloblasts and odontoblasts during the formation of enamel and dentin matrices, respectively. The FRG1 protein was localized in the cytoplasm in the mouse Tooth Germ in vivo, while FRG1 was detected predominantly in the nucleus and faintly in the cytoplasm in mDE6 cells in vitro. In mDE6 cells treated with bone morphogenetic protein 4 (BMP4), the protein expression of FRG1 increased in cytoplasm, suggesting that FRG1 may translocate to the cytoplasm. These findings suggest that FRG1 is involved in the morphogenesis of the Tooth Germ, as well as in the formation of enamel and dentin matrices and that FRG1 may play a role in the odontogenesis in the mouse following BMP4 stimulation.

  • The expression and function of thymosin beta 10 in Tooth Germ development.
    The International journal of developmental biology, 2013
    Co-Authors: Maho Shiotsuka, Tamotsu Kiyoshima, Hiroko Wada, Kana Hasegawa, Kengo Nagata, Hiroaki Fujiwara, Hirotaka Someya, Ichiro Takahashi, Makiko Kihara, Hidetaka Sakai
    Abstract:

    This study presents the expression pattern and functions of thymosin beta 10 (Tbeta10), a Tbeta4 homologue during the development of mouse lower first molars. An in situ signal of Tbeta10 was detected on embryonic day 10.5 (E10.5)-E15.5 mainly in dental mesenchymal cells as well as in dental epithelial cells, while Tbeta4 was expressed in dental epithelial cells. In the late bell stage, preodontoblasts with strong Tbeta10 expression and preameloblasts with strong Tbeta4 expression exhibited face-to-face localization, suggesting that an intimate cell-cell interaction might exist between preodontoblasts and preameloblasts to form dentin and enamel matrices. A strong Tbeta10 signal was found in odontoblasts in the lateral side of the dental pulp and in Hertwig’s epithelial root sheath, thus suggesting that Tbeta10 participates in the formation of the outline of the Tooth root. An inhibition assay using Tbeta10-siRNA in E11.0 mandibles showed significant growth inhibition in the Tooth Germ. The Tbeta10-siRNA-treated E15.0 Tooth Germ also showed significant developmental arrest. The number of Ki67-positive cells significantly decreased in the Tbeta10-siRNA-treated mandibles. The cellular proliferative activity was also significantly suppressed in Tb10-siRNA-treated cultured mouse dental pulpal and epithelial cells. These results indicate that developmental arrest of the Tooth Germ might be caused by a reduction in cell proliferative activity. The stage-specific temporal and spatial expression pattern of Tbeta10 in the developing Tooth Germ is indicative of multiple functions of Tbeta10 in the developmental course from initiation to root formation of the Tooth Germ.

  • Multiple functional involvement of Thymosin beta-4 in Tooth Germ development
    Histochemistry and cell biology, 2012
    Co-Authors: Yukiko Ookuma, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Haruyoshi Yamaza, Hiroko Wada, Kengo Nagata, Hiroaki Fujiwara, Kazuaki Nonaka, Hidetaka Sakai
    Abstract:

    Thymosin beta-4 (Tβ4) is known to be ubiquitously involved in the actin monomer sequestering on the cytoskeleton. Our previous study showed specific temporal and special in situ expression pattern of Tβ4 mRNA in dental epithelial and mesenchymal cells in the developing Tooth Germ of the mouse lower first molar. In this study, we examined the functional implications of Tβ4 in the developmental course of the mouse lower first molar. An inhibition assay using Tβ4 antisense sulfur-substituted oligodeoxynucleotide (AS S-ODN) in cultured embryonic day 11.0 (E11.0) mandibles showed a significant growth inhibition of the Tooth Germ. However, no growth arrest of the cultured E15.0 Tooth Germ was observed by using Tβ4 AS S-ODN. The Tβ4 knockdown led to significantly decreased expression levels of type II/III runt-related transcription factor 2 (Runx2) and nucleolin (Ncl) in the cultured E11.0 mandibles. Since our previous studies proved that the inhibition of type II/III Runx2 and Ncl translations resulted in the developmental arrest of the Tooth Germ in the cultured E11.0 mandible, Tβ4 appears to play roles in Tooth Germ development via the regulation of the type II/III Runx2 and Ncl expressions. Tβ4 knockdown also resulted in decreased secretion of matrix metalloproteinase (Mmp)-2, a reduced cell motility activity and upregulation of E-cadherin in dental epithelial mDE6 cells. These results suggest that Tβ4 plays multiple functional roles in odontogenic epithelial cells in the early stages of Tooth Germ development by regulating the expression of odontogenesis-related genes.