Molecular cytochemical analysis of correlation,between nuclear distribution of transcription factors and transcriptional activation or cell differentiation inductivity

• Project/Area Number: 18590189
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General anatomy (including Histology/Embryology)
• Research Institution: Nagasaki University
• Principal Investigator: IZUMI Shin-ichi Nagasaki University, Graduate School of Biomedical Sciences, Assistant Professor (40264246)
• Co-Investigator(Kenkyū-buntansha): KOMORI Toshihisa Nagasaki University, Graduate School of Biomedical Sciences, Professor (00252677) ; MIYAZAKI Toshihiro Nagasaki University, Graduate School of Biomedical Sciences, Assistant Professor (10174161)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥3,760,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥360,000); Fiscal Year 2007: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2006: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: Transcription factor / Nucleus / Differentiation / Cytochemistry / Runx2 / Gene / Osteoblast / Transgenic / 発生・分化 / 核酸 / 細胞・組織 / 生体分子

Research Abstract

Runx2, a transcription factor plays an important role in cell differentiation of pluripotential undifferentiated mesenchyme into osteoblasts and chondrocytes, and regulates transcription of the target genes by binding to the promoter. Molecular cytochemistry was performed to clarify whether nuclear distribution of Runx2 correlates with the transcription activation and/or DNA binding activity of the transcription factors for the differentiation.
1. When bone morphology protein-2 was introduced to pre-osteoblasts to promote the differentiation by increasing transcription activation of Runx2 gene, Runx2 was strongly stained in the euchromatin area in nuclei.
2. In fetal cartilage of transgenic mice with CoIII-Akt-Tg-GFP, Runx2 appeared in the nuclei of undifferentiated perichondrial cells and proliferating chondrocytes. The staining was increased in the nuclear matrix of mature chondrocytes, and decreased in the differentiated hypertrophy chondrocytes. Nuclear distribution of Runx2 seemed to correlate with DNA binding changes of Runx2 by Akt and chondrocyte differentiation.
3. In tibia of new-born mice of Runx2-Tg, Runx2 and β-catenin appeared in the nuclei of undifferentiated perichondrial cells, and Runx2 was distributed in the nuclear matrix of osteoblasts but not in the osteocytes. In fetal mice of Runx2-/-, neither β-catenin nor osterix was stained in the nuclei of undifferentiated perichondrial cells. Nuclear distribution of Runx2 and β-catenin seemed to be related with the differentiation.
4. When Runx2/GFP were induced in C3H10T1/2 pluripotent undifferentiated fibroblasts, the cytoplasmic processes of the cells were decreased and became short. Immunostaining of actin was decreased and membranous β-catenin aggregated in the cytoplasm. Nuclear matrix distribution of Runx2 seemed to be related with induction of the cell differentiation.

Research Products

• [Journal Article] Localization of specific molecules by electron microscopy (2008)
  • Author(s): Izumi S
  • Journal Title: Journal of Electron Microscopy Technology for Medicine and Biology 21(2) Pages: 167-171
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Localization of specific molecules by electron microscopy (2008)
  • Author(s): Izumi, S
  • Journal Title: Journal of Electron Microscopy Technology for Medicine and Biology 21(2) Pages: 167-171
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Antibody size for immuno-detection of mitochondrial adrenodoxin and adrenodoxin reductase in the rat adrenal tissues treated with aminoglutethimide or 4-aminopyrazolopyrimidine and in the human adrenal with hyperaldosteroinism (2007)
  • Author(s): Izumi S
  • Journal Title: Journal of Electron Microscopy Technology forMedicine and Biology 21(1) Pages: 71-75
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Runx2 determines bone maturity and turnover rate in postnatal bone development and is involved in bone loss in estrogen deficiency (2007)
  • Author(s): Maruyama Z
  • Journal Title: Developmental Dynamics 236(7) Pages: 1876-1890
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] 免疫電顕におけるトラブルシューティング (2007)
  • Author(s): 和泉 伸一
  • Journal Title: 顕微鏡 42(1) Pages: 16-18
  • NAID: 10018912844
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Antibody size for immuno-detection of mitochondrial adrenodoxin and adrenodoxin reductase in the rat adrenal tissues treated with aminoglutethimide or 4-aminopyrazolopyrimidine and in the human adrenal with hyperaldosteroinism (2007)
  • Author(s): Izumi, S
  • Journal Title: Journal of Electron Microscopy Technology for Medicine and Biology 21(1) Pages: 71-75
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Runx2 determines bone maturity and turnover rate in postnatal bone development and is involved in bone loss in estrogen deficiency (2007)
  • Author(s): Maruyama, Z., Yoshida, CA., Furuichi, T., Amizuka, N., Ito, M., Fukuyama, R., Miyazaki, T., Kitaura, H., Nakamura, K., Fujita, T., Kanatani, N., Moriishi, T., Yamana, K., Liu, W., Kawaguchi, H., Nakamura, K., Komori, T
  • Journal Title: Developmental Dynamics 236(7) Pages: 1876-1890
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Trouble-shooting for immuno-electron microscopy.(In Japanese) (2007)
  • Author(s): Izumi, S
  • Journal Title: Kenbikyou 42(1) Pages: 16-18
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Runx2 determines bone maturity and turnover rate in postnatal bone development and is involved in bone loss in estrogen deficiency (2007)
  • Author(s): Maruyama Z.
  • Journal Title: Developmental Dynamics 236(7) Pages: 1876-1890
  • Peer Reviewed
• [Journal Article] Antibody size for immuno-detection of mitochondrial adrenodoxin and adrenodoxin reductase in the rat adrenal tissues treated with aminoglute thimide or 4-aminopyrazolopyrimidine and in the human adrenal with hyperaldosteroinism (2007)
  • Author(s): Izumi S
  • Journal Title: Journal of Electron Microscopy Technology for Medicine and Biology 21(1) Pages: 71-75
  • Peer Reviewed
• [Journal Article] Localization of specific molecules by electron microscopy (2007)
  • Author(s): Izumi S
  • Journal Title: Journal of Electron Microscopy Technology for Medicine and Biology 21(2) Pages: 1-5
  • Peer Reviewed
• [Journal Article] Localization of specific genes,transcripts and transcription factors in the cellorganelles (2006)
  • Author(s): Izumi S
  • Journal Title: Journal of Electron Microscopy Technology for Medicine and Biology 20(2) Pages: 101-102
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Diethylstilbestrol increases the densily of prolactin cells in male mouse pituitary by inducing proliferation of prolactin cells and transdifferentiation of gonadotropic cells (2006)
  • Author(s): Shukuwa K
  • Journal Title: Histochemistry and Cell Biology 126 Pages: 111-123
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] 選択的透過性を失ったミトコンドリアの電顕酵素抗原法による検出 (2006)
  • Author(s): 和泉 伸一
  • Journal Title: 医学生物学電子顕微鏡技術学会誌 20(1) Pages: 30-30
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 象牙芽細胞の分化過程におけるRunx2の作用に関する組織学的解析 (2006)
  • Author(s): 宮崎 敏博
  • Journal Title: Journal of Oral Biosciences 48・Suppl.
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Cbfβ regulates Runx2 function isoform-dependency in postnatal bone development (2006)
  • Author(s): Kanatani N
  • Journal Title: Developmental Biology 296 Pages: 48-61
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Localization of specific genes, transcripts and transcription factors in the cell organelles (2006)
  • Author(s): Izumi, S
  • Journal Title: Journal of Electron Microscopy Technology for Medicine and Biology 20(2) Pages: 101-102
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Diethylstilbestrol increases the density of prolactin cells in male mouse pituitary by inducing proliferation of prolactin cells and transdifferentiation of gonadotropic cells (2006)
  • Author(s): Shukuwa, K., Izumi, S., Hishikawa, Y., Ejima, K., Inoue, S., Muramatsu, M., Ouchi, Y., Kitaoka, T., Koji, T
  • Journal Title: Histochemistry and Cell Biology 126 Pages: 111-123
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Electron microscopic immunocytochemical detection of mitochondria in a permeability transition.(In Japanese) (2006)
  • Author(s): Izumi, S
  • Journal Title: Journal of Electron Microscopy Technology for Medicine and Biology 20, 30
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Histological analysis concerning Runx2 activity on differentiation processes of odontoblasts 48 Suppl.(In Japanese) (2006)
  • Author(s): Miyazaki, T., Eguchi, T., Moriishi, T., Kanatani, N., Izumi, S., Toyosawa, S., Komori, T
  • Journal Title: Journal of Oral Biosciences 154(468)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cbfβregulates Runx2 function isoform-dependently in postnatal bone development (2006)
  • Author(s): Kanatani, N., Fujita, T., Fukuyama, R., Liu, W., Yoshida, CA., Moriishi, T., Yamana, K., Miyazaki, T., Toyosawa, S., Koji, T
  • Journal Title: Developmental Biology 296 Pages: 48-61
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Localization of specific genes, transcripts and transcription factors in the cell organelles. (2006)
  • Author(s): Izumi S
  • Journal Title: Journal of Electron Microscopy Technology for Medicine and Biology 20・2 Pages: 101-102
• [Journal Article] Diethylstilbestrol increases the density of prolactin cells in male mouse pituitary by inducing proliferation of prolactin cells and transdifferentiation of gonadotropic cells. (2006)
  • Author(s): Shukuwa K
  • Journal Title: Histochemistry and Cell Biology 126 Pages: 111-123
• [Journal Article] 選択的透過性を失ったミトコンドリアの電顕酵素抗原法による検出 (2006)
  • Author(s): 和泉伸一
  • Journal Title: 医学生物学電子顕微鏡技術学会会誌 20・1 Pages: 30-30
• [Journal Article] 象牙芽細胞の分化過程におけるRunx2の作用に関する組織学的解析 (2006)
  • Author(s): 宮崎敏博
  • Journal Title: Journal of Oral Biosciences 48・Suppl.
• [Journal Article] Cbfβ regulates Runx2 function isoform-dependency in postnatal bone development. (2006)
  • Author(s): Kanatani N
  • Journal Title: Developmental Biology 296 Pages: 48-61
• [Presentation] 転写因子の核内分布と遺伝子発現 (2008)
  • Author(s): 和泉 伸一
  • Organizer: 第113回日本解剖学会総会・全国学術集会
  • Place of Presentation: 大分
  • Year and Date: 2008-03-27
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Nuclear distribution of transcription factors and gene expression(In Japanese) (2008)
  • Author(s): Izumi, S
  • Organizer: The 113th annual meeting of Japanese society of anatomy
  • Place of Presentation: Oita(Symposium)
  • Year and Date: 2008-03-27
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 電顕in situ hybridizationによる核小体rRNAの局在 (2007)
  • Author(s): 末松 貴史
  • Organizer: 日本解剖学会第63回九州支部学術集会
  • Place of Presentation: 長崎
  • Year and Date: 2007-10-20
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 骨芽細胞特異的Bcl-2トランスジェニックマウスを用いた、骨細胞による骨量調節の検証 (2007)
  • Author(s): 森石 武史
  • Organizer: 日本解剖学会第63回九州支部学術集会
  • Place of Presentation: 長崎
  • Year and Date: 2007-10-20
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Localization of nucleolar rRNA by electron microscopic in situ hybridization(In Japanese) (2007)
  • Author(s): Suematsu, T., Izumi, S., Hishikawa, Y., Koji, T
  • Organizer: The 63rd annual meeting of Japanese society of anatomy(Kyushu region meeting)
  • Place of Presentation: Nagasaki
  • Year and Date: 2007-10-20
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Verification of bone mass regulation by osteocytes with use of osteoblast targeted Bcl-2 transgenic mice(In Japanese) (2007)
  • Author(s): Moriishi, T., Miyazaki, T., Izumi, S., Kanatani, N., Komori, T
  • Organizer: The 63rd annual meeting of Japanese society of anatomy(Kyushu region meeting)
  • Place of Presentation: Nagasaki
  • Year and Date: 2007-10-20
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 骨芽細胞特異的Bc1-2トランスジェニックマウスを用いた、骨細胞による骨量調節の検証 (2007)
  • Author(s): 森石 武史
  • Organizer: 日本解剖学会第63回九州支部学術集会
  • Place of Presentation: 長崎市(長崎大学)
  • Year and Date: 2007-10-20
• [Presentation] 電顕in situ hybridizationによる核小体rRNAの局在 (2007)
  • Author(s): 末松 貴史
  • Organizer: 日本解剖学会第63回九州支部学術集会
  • Place of Presentation: 長崎市(長崎大学)
  • Year and Date: 2007-10-20
• [Presentation] LR White Resinを用いた倒立法埋法の検討 (2007)
  • Author(s): 末松 貴史
  • Organizer: 医学生物学電子顕微鏡技術学会第23回学術講演会
  • Place of Presentation: 北九州
  • Year and Date: 2007-05-20
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] A study of the inverting-embedding method by using LR White Resin(In Japanese) (2007)
  • Author(s): Suematsu, T., Izumi, S
  • Organizer: The 23rd annual meeting of society of electron microscopy technology for medicine and biology
  • Place of Presentation: Kitakyushu
  • Year and Date: 2007-05-20
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Post-embedding電顕in situ hybridization法 (2007)
  • Author(s): 末松 貴史
  • Organizer: 医学生物学電子顕微鏡技術学会第23回学術講演会
  • Place of Presentation: 北九州(産業医科大学)
  • Year and Date: 2007-05-20
• [Presentation] LR White Resinを用いた倒立法埋法の検討 (2007)
  • Author(s): 末松 貴史
  • Organizer: 医学生物学電子顕微鏡技術学会第23回学術講演会
  • Place of Presentation: 北九州(産業医科大学)
  • Year and Date: 2007-05-20
• [Presentation] Post-embedding電顕in situ hybridization法 (2007)
  • Author(s): 末松 貴史
  • Organizer: 医学生物学電子顕微鏡技術学会第23回学術講演会
  • Place of Presentation: 北九州
  • Year and Date: 2007-05-19
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Post-embedding electron microscopic in situ hybridization(In Japanese) (2007)
  • Author(s): Suematsu, T., Izumi, S., Hishikawa, Y., Koji, T
  • Organizer: The 23rd annual meeting of society of electron microscopy technology for medicine and biology
  • Place of Presentation: Kitakyushu
  • Year and Date: 2007-05-19
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Overexpression of Runx2 in mesenchymal cells enhallced hone formation but inhibited cartiage formation and limb development (2007)
  • Author(s): Yoshida CA
  • Organizer: The 29th Annual Meeting of ASBMR(the American Society of Bone and Mineral Metabolism)
  • Place of Presentation: Honolulu
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Over-expression of Runx2 in mesenchymal cells enhanced bone formation but inhibited cartilage formation and limb development (2007)
  • Author(s): Yoshida, CA., Maeno, T., Kanatani, N., Izumi, S., Komori, T
  • Organizer: The 29th Annual Meeting of ASBMR(the American Society of Bone and Mineral Metabolism)
  • Place of Presentation: Honolulu, USA
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 電顕ISHによる核小体での18S rRNA及び28S rRNAの局在検討 (2006)
  • Author(s): 末松 貴史
  • Organizer: 第48回日本顕微鏡学会九州支部総会・学術講演会
  • Place of Presentation: 長崎
  • Year and Date: 2006-12-02
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Localization of 18S rRNA and 28S rRNA in nucleolus by electron microscopic ISH(In Japanese) (2006)
  • Author(s): Suematsu, T., Izumi, S., Hishikawa, Y., Koji, T
  • Organizer: The 48th annual meeting of Japanese society of microscopy(Kyushu region meeting)
  • Place of Presentation: Nagasaki
  • Year and Date: 2006-12-02
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 抗体の大きさと副腎組織切片におけるミFコンドリア内アドレノドキシンの免疫電顕による検出度の関係 (2006)
  • Author(s): 和泉 伸一
  • Organizer: 医学生物学電子顕微鏡技術学会第22回学術講演会
  • Place of Presentation: 浜松
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Subnucleolar localization of ribosomal RNA gene transcripts in mouse Sertoli cell nucleolus by light and electron microscopic in situ hybridization (2006)
  • Author(s): Koji T
  • Organizer: The 16th International Microscopy Congress
  • Place of Presentation: Sapporo
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 象牙芽細胞の分化過程におけるRunx2の作用に関する組織学的解析 (2006)
  • Author(s): 宮崎 敏博
  • Organizer: 第48回歯科基礎医学会学術大会
  • Place of Presentation: 横浜
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Relationship between antibody size and electron microscopic immuno-detection of mitochondrial adrenodoxin in the adrenals,(In Japanese) (2006)
  • Author(s): Izumi, S
  • Organizer: The 22nd annual meeting of society of electron microscopy technology for medicine and biology
  • Place of Presentation: Hamamatsu
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Subnucleolar localization of ribosomal RNA gene transcripts in mouse Sertoli cell nucleolus by light and electron microscopic in situ hybridization (2006)
  • Author(s): Koji, T., Hishikawa, Y., Izumi, S., Suematsu, T
  • Organizer: The 16th International Microscopy Congress
  • Place of Presentation: Sapporo, Japan(Symposium)
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Histological analysis concerning Runx2 activity on differentiation processes of odontoblasts(In Japanese) (2006)
  • Author(s): Miyazaki, T., Eguchi, T., Moriishi, T., Kanatani, N., Izumi, S., Toyosawa, S., Komori, T
  • Organizer: The 48th annual meeting of society of oral basic medicine
  • Place of Presentation: Yokohama
  • Description: 「研究成果報告書概要(欧文)」より