Study on mechanisms of odontoblasts processes formation and dentin development

2009 Fiscal Year Final Research Report

• Project/Area Number: 19390466
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Morphological basic dentistry
• Research Institution: Iwate Medical University
• Principal Investigator: HARADA Hidemitsu Iwate Medical University, 歯学部, 教授 (70271210)
• Co-Investigator(Renkei-kenkyūsha): ISHIZEKI Kiyoto 岩手医科大学, 歯学部, 准教授 (50057775) ; FUJIWARA Naoki 岩手医科大学, 歯学部, 講師 (20190100) ; KAGIYA Tadayoshi 岩手医科大学, 歯学部, 助教 (30405774)
• Project Period (FY): 2007 – 2009
• Keywords: 歯の発生 / 象牙芽細胞 / リアルタイムイメージング / RECK / ROCK / 細胞極性 / マトリックスメタロプロテアーゼ / アメロゲニン

Research Abstract

We designed the real time imaging system in a slice tooth germ culture of actin promoter GFP transgenic mouse. Using the technique, we could capture the cell division and movement of mesenchymal cells in the dental papilla and demonstrate the formation of odontoblasts processes. We examined the role of Rho-associated kinase (ROCK) in odontoblasts differentiation using an organ culture of mouse incisor and mouse dental mesenchaml cell line. ROCK inhibitors and ROCK siRNA inhibited actin polymerization and E-cadherin expression on the cytoplasmic membrane. ROCK plays an important role in maintenance of cellular polarization through regulations of cytoskeleton and cell adhesion in the odontoblasts differentiation. Odotoblasts expressed the amelogenin, MMP inhibitors and MMP. Our data suggested that these factors play an important role in the maintenance of dentin canal.

Research Products

• [Journal Article] Identification of a suppressor element in the amelogenin promoter. (2010)
  • Author(s): Xu L., Matsumoto A., Sasaki A., Harada H., Taniguchi A.
  • Journal Title: J. Dent. Res 89(3) Pages: 246-51
• [Journal Article] Reduction of Egf signaling decides transition from crown to root in the development of mouse molars. (2009)
  • Author(s): ujiwara N, Akimoto T, Otsu K, Kagiya T, Ishizeki K, Harada H.
  • Journal Title: J Exp Zoolog B Mol Dev Evol 312B(5) Pages: 486-94
• [Journal Article] MAPK mediates Hsp25 signaling in incisor development. (2009)
  • Author(s): Lee MJ, Cai J, Kwak SW, Cho SW, Harada H, Jung HS
  • Journal Title: Histochem Cell Biol 131(5) Pages: 593-603
• [Journal Article] Secreted Frizzled Related Protein (sFRP)-2 Inhibits Bone Formation and Promotes Cell Proliferation in Ameloblastoma. (2009)
  • Author(s): Gulsan AS, Inoue M, Harada H, Rodriguez AP, Tamamura R, Tsujigiwa H, Borkosky SS, Gunduz M, Nagatsuka H
  • Journal Title: Oral Oncol 45(10) Pages: 856-60
• [Journal Article] Gene expression analyses of human macrophage phagocytizing sub-micro titanium particles by allergy DNA chip (Genopal). (2009)
  • Author(s): Taira M, Nezu T, Sasaki M, Kimura S, Kagiya T, Harada H, Narushima T, Araki Y
  • Journal Title: Biomed Mater Eng 19(1) Pages: 63-70
• [Journal Article] Expression of osteogenic proteins during the intrasplenic transplantation of Meckel's chondrocytes: A histochemical and immunohistochemical study. (2009)
  • Author(s): Ishizeki K, Kagiya T, Fu, jiwara N, Otsu K, Harada H
  • Journal Title: Arch Histol Cytol 72(1) Pages: 1-12
• [Journal Article] Apc inhibition of Wnt signaling regulates supernumerary tooth formation during embryogenesis and throughout adulthood. (2009)
  • Author(s): Wang XP, 0'Connell DJ, Lund JJ, Saadi I, Kuraguchi M, Turbe-Doan A, Cavallesco R, Kim H, Park PJ, Harada H, Kucherlapati R, Maas RL
  • Journal Title: Development 136(11) Pages: 1939-49
• [Journal Article] Preparation and in vivo evaluation of apatite/collagen packed composite by alternate immersion method and Newton press. (2009)
  • Author(s): Taira, M., Nezu, T., Sasaki, K., Saitoh, S., Kagiya, T., Harada, H., Takada, Y., Araki, Y.
  • Journal Title: J. Biomed. Mater. Res. B Appl. Biomater. 90 Pages: 566-573
• [Journal Article] Microscopic observations and inflammatory cytokine productions of human macrophage phagocytising submicron titanium particles. (2009)
  • Author(s): Taira, M., Kagiya, T., Harada, H., Sasaki, M., Kimura, S., Narushima, T., Nezu, T., Araki, Y.
  • Journal Title: J. Mater. Sci. Mater. Med. 20 Pages: 10. 1007/s10856-009-3834
• [Journal Article] Critical role of heparin binding domains of ameloblastin for dental epithelium cell adhesion and ameloblastoma proliferation. (2009)
  • Author(s): Sonoda A, Iwamoto T, Nakamura T, Fukumoto E, Yoshizaki K, Yamada A, Arakaki M, Harada H, Nonaka K, Nakamura S, Yamada Y, Fukumoto S.
  • Journal Title: J Biol. Chem. 2;284(40) Pages: 27176-84
• [Journal Article] The epithelial-mesenchymal interaction plays a role in the maintenance of the stem cell niche of mouse incisors via Fgf10 and Fgf9 signaling. (2008)
  • Author(s): Yokohama-Tamaki T, Fujiwara N, Shibata S, Wakisaka S, Harada H
  • Journal Title: Open Biotechnol J 2 Pages: 111-115
• [Journal Article] Hydroxyapatite-and amelogenin protein-induced expression of mineralization-related genes in a dental epithelial cell line. (2008)
  • Author(s): Xu L., Harada H., Ikoma T., Taniguchi A
  • Journal Title: Open Biotechnol J 2 Pages: 116-120
• [Journal Article] BMP2-induced gene profiling in dental epithelial cell line. (2008)
  • Author(s): Keiko Miyoshi, Hideya Nagata, Taigo Horiguchi, Kaori Abe, Ivan Arie Wahyudi, Yoshinobu Baba, Hidemitsu Harada, Takafumi Noma
  • Journal Title: J. Med. Invest. 55 Pages: 216-226
• [Journal Article] The relationship between the cusp pattern and plural stem cell compartments in guinea pig cheek teeth by chasing BrdU-labeling. (2008)
  • Author(s): Hashimoto E, Nakakura-Ohshima K, Kenmotsu S, Suzuki H, Nakasone N, Saito C, Harada H, Ohshima H
  • Journal Title: Arch Histol Cytol 71(5) Pages: 317-32
• [Journal Article] Neurotrophic factor NT-4 regulates ameloblastin expression via full-length TrkB. (2008)
  • Author(s): Yoshizaki K, Yamamoto S, Yamada A, Yuasa K, Iwamoto T, Fukumoto E, Harada H, Saito M, Nakasima A, Nonaka K, Yamada Y, Fukumoto S
  • Journal Title: J Biol Chem 283(6) Pages: 3385-3391
• [Journal Article] The effects of LAMP1 and LAMP3 on M180 amelogenin uptake, localization and amelogenin mRNA induction by amelogenin protein. (2008)
  • Author(s): Xu L, Harada H, Taniguchi A
  • Journal Title: J Biochem 144(4) Pages: 531-537
• [Journal Article] Establishment of immortalized dental follicle cells for generating periodontal ligament in vivo. (2007)
  • Author(s): Yokoi, T., Saito, M., Kiyono, T., Iseki, S., Kosaka, K., Nishida, E., Tsubakimoto, T., Harada, H., Eto, K., Noguchi, T., Teranaka, T.
  • Journal Title: Cell Tissue Res 327(2) Pages: 301-11
• [Journal Article] Effect of BMP-2 on gene expression of enamel matrix proteins s at the dental epithelial cell line. (2007)
  • Author(s): iming Xu, Ryoya Takahashi, Hidemitsu Harada, Akiyoshi Taniguchi
  • Journal Title: Open Biotechnol J 1 Pages: 18-20
• [Journal Article] Immunocytochemical and biochemical detection of EMMPRIN in the rat tooth germ: differentiation-dependent co-expression with MMPs and co-localization with caveolin-1 in membrane rafts of dental epithelial cells. (2007)
  • Author(s): Schwab W, HaradaH, Goetz W, Nowicki M, Witt M, Kasper M, Barth K
  • Journal Title: Histochem Cell Biol 128(3) Pages: 195-203
• [Presentation] Rho.キナーゼによるエナメル芽細胞の極性制御機構 (2009)
  • Author(s): 大津圭史、藤原尚樹、石関清人、鍵谷忠慶、佐々木憲明, 原田英光
  • Organizer: 第51回歯科基礎医学会学術大会
  • Place of Presentation: 新潟
  • Year and Date: 20090910-20090911
• [Presentation] GFPマウスを用いたメッケル軟骨細胞の形質転換のイメージング解析 (2009)
  • Author(s): 石関清人、大津圭史、鍵谷忠慶、藤原尚樹、原田英光
  • Organizer: 第51回歯科基礎医学会学術大会
  • Place of Presentation: 新潟
  • Year and Date: 20090910-20090911
• [Presentation] エナメル芽細胞分化課程における細胞間結合の役割 (2009)
  • Author(s): 山田亜矢、岩本勉、鈴木宏冶、中村卓志、原田英光、斉藤正寛、福本敏
  • Organizer: 第51回歯科基礎医学会学術大会
  • Place of Presentation: 新潟
  • Year and Date: 20090910-20090911
• [Presentation] ヘルトヴィッヒ上皮鞘形成過程におけるサービカルループでの細胞動態 (2009)
  • Author(s): 原田英光、石関清人、大津圭史、鍵谷忠慶、藤原尚樹
  • Organizer: 第51回歯科基礎医学会学術大会
  • Place of Presentation: 新潟
  • Year and Date: 20090910-20090911
• [Presentation] 天然低分子化合物を用いた歯根・歯周組織再生. (2009)
  • Author(s): 米澤貴之, 太田正人, 藤原尚樹, 車柄允, 照屋俊明, 鍵谷忠慶, 原田英光, 矢ヶ崎一三, 永井和夫, 禹済泰
  • Organizer: 第21回日本動物細胞工学会
  • Place of Presentation: つくば
  • Year and Date: 20090700
• [Presentation] 歯の再生を目的としたiPS細胞の上皮 (2009)
  • Author(s): 大津圭史、石関清人、鍵谷忠慶、藤原尚樹、原田英光
  • Organizer: 間葉細胞への分化誘導と分離培養第114回日本解剖学会総会・全国学術大会
  • Place of Presentation: 岡山
  • Year and Date: 20090328-20090330
• [Presentation] 歯胚組織中の細胞動態を観察するためのimaging systemの開発第114回日本解剖学会総会 (2009)
  • Author(s): 藤原尚樹、大津圭史、秋元義、鍵谷忠慶、石関清人、原田英光
  • Organizer: 全国学術大会
  • Place of Presentation: 岡山
  • Year and Date: 20090328-20090330
• [Presentation] 銅イオンのマクロファージへ及ぼす細胞障害作用について第114回日本解剖学会総会 (2009)
  • Author(s): 鍵谷忠慶、秋元義、大津圭史、藤原尚樹、石関清人、原田英光, 吉田康夫、平雅之
  • Organizer: 全国学術大会
  • Place of Presentation: 岡山
  • Year and Date: 20090328-20090330
• [Presentation] メッケル軟骨の形質転換の意味するもの (2008)
  • Author(s): 石関清人、鍵谷忠慶、藤原尚樹、原田英光
  • Organizer: 第50回歯科基礎医学会学術大会
  • Place of Presentation: 昭和大学(東京)
  • Year and Date: 20080923-20080925
• [Presentation] 歯冠から歯根形成への移行過程におけるEGF signalingの役割 (2008)
  • Author(s): 藤原尚樹、大津圭史、秋元義、鍵谷忠慶、石関清人、原田英光
  • Organizer: 第50回歯科基礎医学会学術大会
  • Place of Presentation: 昭和大学(東京)
  • Year and Date: 20080923-20080925
• [Presentation] 高濃度銅イオンによるマクロファージの細胞形態変化と障害作用の評価 (2008)
  • Author(s): 平雅之, 鍵谷忠慶, 原田英光, 佐々木実, 木村重信
  • Organizer: 第50回歯科基礎医学会学術大会
  • Place of Presentation: 昭和大学(東京)
  • Year and Date: 20080923-20080925
• [Presentation] エナメル上皮腫における細胞生物学的特徴とその病態について (2008)
  • Author(s): 光安岳志、川野真太郎、中尾祐、ヘンダーミンライファ、原田英光、中村典史、中村誠司
  • Organizer: 日本口腔科学会学術集会
  • Place of Presentation: 福岡
  • Year and Date: 20080417-20080418
• [Presentation] ヘルトビィッヒ上皮鞘細胞の不死化 (2008)
  • Author(s): 秋元義、藤原尚樹、鍵谷忠慶、石関清人、原田英光
  • Organizer: 第113回日本解剖学会総会、全国学術集会
  • Place of Presentation: 大分大学(大分)
  • Year and Date: 20080327-20080329
• [Presentation] マウス切歯の組織幹細胞を用いた歯の再生 (2008)
  • Author(s): 原田英光、藤原尚樹、鍵谷忠慶、石関清人
  • Organizer: 第7回日本再生医療学会シンポジウム
  • Place of Presentation: 名古屋
  • Year and Date: 20080313-20080314
• [Presentation] 破骨細胞の細胞死におけるカルパインの関与について (2008)
  • Author(s): 鍵谷忠慶、佐々木憲明、石関清人、藤原尚樹、原田英光
  • Organizer: 岩手医科大学歯学会第66回例会
  • Place of Presentation: 岩手医大(盛岡)
  • Year and Date: 2008-07-05
• [Presentation] 破骨細胞の細胞死におけるカルパインの関与について (2008)
  • Author(s): 鍵谷忠慶、佐々木憲明、石関清人、藤原尚樹、原田英光
  • Organizer: 岩手医科大学歯学会第66回例
  • Place of Presentation: 岩手医大(盛岡)
  • Year and Date: 2008-07-05
• [Presentation] マウス切歯幹細胞を用いた歯の再生 (2008)
  • Author(s): 原田英光、鍵谷忠慶、藤原尚樹、石関清人
  • Organizer: 第113回日本解剖学会全国学術集会シンポジウム
  • Place of Presentation: 大分大学(大分市)
  • Year and Date: 2008-03-29
• [Presentation] 石関清人歯の幹細胞の維持機構の解明から歯の再生への展開第30回日本分子生物学会 (2007)
  • Author(s): 原田英光、藤原尚樹、鍵谷忠慶
  • Organizer: 第80回日本生化学会
  • Place of Presentation: 横浜
  • Year and Date: 20071211-20071215
• [Presentation] 原田英光歯の発生時におけるアメロゲニンの大量発現メカニズム第30回日本分子生物学会 (2007)
  • Author(s): 谷口彰良、徐麗明
  • Organizer: 第80回日本生化学会
  • Place of Presentation: 横浜
  • Year and Date: 20071211-20071215
• [Presentation] ラットアメロゲニン遺伝子の細胞特異的転写調節 (2007)
  • Author(s): 徐麗明、松本阿佐子、原田英光、谷口彰良
  • Organizer: 第30回日本分子生物学会・第80回日本生化学会
  • Place of Presentation: 横浜
  • Year and Date: 20071211-20071215
• [Presentation] エナメル上皮細胞と間葉細胞との共培養によるエナメルマトリックス遺伝子の発現変化 (2007)
  • Author(s): 松本阿佐子, 徐麗明, 斉藤正寛, 原田英光, 谷口彰良
  • Organizer: 第30回日本分子生物学会・第80回日本生化学会
  • Place of Presentation: 横浜
  • Year and Date: 20071211-20071215
• [Presentation] Mechanisms on maintenance of dental stem cells and how to make a biotooth. (2007)
  • Author(s): H Harada N Fujiwara, T Yokohama-Tamaki, T Kagiya, Y Tabata, K Ishizaki
  • Organizer: lst ABMC
  • Place of Presentation: Tsukuba, Japan
  • Year and Date: 20071206-20071208
• [Presentation] Effects of amelogenin and Hap on dental epithelial cell differentiation and enamel formation. (2007)
  • Author(s): A Taniguchi, L Xu, H Harada
  • Organizer: lst ABMC
  • Place of Presentation: Tsukuba, Japan
  • Year and Date: 20071206-20071208
• [Presentation] Mechanisms on maintenance of dental stem cells and how to make a biotooth. (2007)
  • Author(s): Harada, H., Fujiwara, N., Y-Tamaki, T., Kagiya, T., Tabata, Y., Ishizek.K.
  • Place of Presentation: Tsukuba, Japan
  • Year and Date: 20071206-20071208
• [Presentation] Molecular Mechanisms of Amelogenin over Expression and Enamel Formation at Tooth Development. (2007)
  • Author(s): Taniguchi, A., Xu, L., Harada, H.
  • Organizer: 1st ABMC
  • Place of Presentation: Tsukuba, Japan
  • Year and Date: 20071206-20071208
• [Presentation] Increased ameloblastin expression in dental epithelial cell line by co-culture with mesenchymal cell line. (2007)
  • Author(s): Matsumoto, A., Xu, L., Saito, M., Harada, H., Taniguchi, A.
  • Organizer: 1st ABMC
  • Place of Presentation: Tsukuba, Japan
  • Year and Date: 20071206-20071208
• [Presentation] 間葉系細胞との共培養によるエナメル上皮細胞の分化マーカー遺伝子発現の変化 (2007)
  • Author(s): 松本阿佐子, 徐麗明, 斉藤正寛, 原田英光, 谷口彰良
  • Organizer: 第29回日本バイオマテリアル学会
  • Place of Presentation: 大阪
  • Year and Date: 20071126-20071127
• [Presentation] ヘルトビッヒの上皮鞘(HERS)形成過程の新規仮説と歯根発生メカニズム (2007)
  • Author(s): 原田英光, 鍵谷忠慶, 藤原尚樹石関清人
  • Organizer: 第49回歯科基礎医学会学術大会サテライトシンポジウム
  • Place of Presentation: 札幌
  • Year and Date: 20070829-20070831
• [Presentation] 幹細胞を用いた歯科再生研究の現状と臨床への展望と課題. (2007)
  • Author(s): 原田英光
  • Organizer: 第24回日本顎顔面補綴学会総会
  • Place of Presentation: 盛岡
  • Year and Date: 20070720-20070721
• [Presentation] EGF prevents formation of Hertwig' s epithelial root sheath during developing mouse molar tooth in vitro. (2007)
  • Author(s): Naoki Fujiwara, Tadayoshi Kagiya, Kiyoto Ishizeki, Hidemitsu Harada
  • Organizer: 9th International Conference on Tooth Morphogenesis and Differentiation 2007
  • Place of Presentation: Zurich, Switzerland
  • Year and Date: 20070409-20070809
• [Presentation] A role of Wnt5a in continuously growing mice incisors. (2007)
  • Author(s): Kagiya, T., Fujiwara, N., Ishizeki, K., J Xiao, Harada, H.
  • Organizer: 9th International Conference on Tooth Morphogenesis and Differentiation
  • Place of Presentation: Zurich, Switzerland
  • Year and Date: 20070409-20070809
• [Presentation] The exon 6ABC region of amelogenin mRNA contribute to increased levels of amelogenin mRNA through amelogenin protein-enhanced mRNA stabilization. (2007)
  • Author(s): Xu L., Harada, H., Taniguchi, A.
  • Organizer: 9th International Conference on Tooth Morphogenesis and Differentiation
  • Place of Presentation: Zurich, Switzerland
  • Year and Date: 20070409-20070809
• [Presentation] Reuptake of extracellular amelogenin by dental epithelial cells results in increased levels of amelogenin mRNA through enhanced mRNA stabilization. (2007)
  • Author(s): Taniguchi, A., Xu, L., Harada, H.
  • Organizer: 9th International Conference on Tooth Morphogenesis and Differentiation
  • Place of Presentation: Zurich, Switzerland
  • Year and Date: 20070409-20070809
• [Presentation] Gap junctional communication regulates ameloblast differentiation. (2007)
  • Author(s): Yamada, A., Fukumoto, E.Yoshizaki, K., Yuasa, K., Yamamoto, S., Iwamoto, T., Furukawa, S., Harada, H., Saito, M., Nonaka, K., Fukumoto, S
  • Organizer: 9th International Conference on Tooth Morphogenesis and Differentiation
  • Place of Presentation: Zurich, Switzerland
  • Year and Date: 20070409-20070809
• [Presentation] Role of P38 kinase and JNK in tooth development. (2007)
  • Author(s): Yamaguchi, Y., Ichioka, H., Toyosawa, S., Harada, H., Nishimura, R., T Yoneda T.
  • Organizer: 9th International Conference on Tooth Morphogenesis and Differentiation
  • Place of Presentation: Zurich, Switzerland
  • Year and Date: 20070409-20070809
• [Presentation] Fgf-9 play a role for the maintenance of stem cell niche via Fgf-10 expression in the mouse incisors. (2007)
  • Author(s): Yokohama-Tamaki, T., Shibata, S., Wakisaka, S., Harada, H.
  • Organizer: 9th International Conference on Tooth Morphogenesis and Differentiation
  • Place of Presentation: Zurich, Switzerland
  • Year and Date: 20070409-20070809
• [Presentation] Neurotrophic factor NT-4 regulates ameloblastin expression. (2007)
  • Author(s): Yoshizaki, K., Yamada, A., Yuasa, K., Yamamoto, S., Iwamoto, T., Harada, H., Saito, M., Fukumoto, E., Nonaka, K., Fukumoto, S.
  • Organizer: 9th International Conference on Tooth Morphogenesis and Differentiation
  • Place of Presentation: Zurich, Switzerland
  • Year and Date: 20070409-20070809
• [Presentation] メッケル軟骨細胞の脾臓内移植からみた細胞の形質転換 (2007)
  • Author(s): 石関清人, 原田英光, 鍵谷忠慶, 藤原尚樹
  • Organizer: 第49回歯科基礎医学会学術大会・総会
  • Place of Presentation: 札幌市
  • Year and Date: 2007-08-30
• [Presentation] ヘルトビッヒの上皮鞘(HERS)形成過程の新規仮説と歯根発生メカニズム (2007)
  • Author(s): 原田英光, 鍵谷忠慶, 藤原尚樹, 石関清人
  • Organizer: 第49回歯科基礎医学会学術大会サテライトシンポジウム
  • Place of Presentation: 札幌市
  • Year and Date: 2007-08-29
• [Presentation] メッケル軟骨の局所形態発生とその運命 (2007)
  • Author(s): 石関清人、鍵谷忠慶、藤原尚樹、原田英光
  • Organizer: 第112回日本解剖学会全国学術集会シンポジウム
  • Place of Presentation: 大阪市
  • Year and Date: 2007-03-28
• [Presentation] Calpain阻害剤を用いた破骨細胞のアポトーシス抑制第112回日本解剖学会総会 (2007)
  • Author(s): 鍵谷忠慶、佐々木憲明、石関清人、藤原尚樹、原田英光
  • Organizer: 全国学術集会
  • Place of Presentation: 大阪市
  • Year and Date: 2007-03-28
• [Presentation] Wnt5aノックアウトマウス歯胚の組織学所見について (2007)
  • Author(s): 鍵谷忠慶、藤原尚樹、石関清人、原田英光
  • Organizer: 岩手医科大学歯学会第36回例会
  • Place of Presentation: 盛岡市
  • Year and Date: 2007-02-24
• [Remarks]
  • URL: http://oralhist.iwate-med.ac.jp/