1993 年度実績報告書

骨折治癒の分子病理学的解析

研究課題

研究課題/領域番号	04670206
研究機関	大阪大学
研究代表者	野村慎太郎大阪大学, 医学部, 助教授 (80159087)
キーワード	骨折治癒 / BMP-4 / 遺伝子発現 / 骨基質 / オステオポンチン / 仮骨 / 発生過程 / 石灰化
研究概要	骨折治癒過程において出現する仮骨の構成蛋白としては、成熟骨と違ってオステオポンチン、オステオネクチンが多量に含まれることが判明した。この現象はBMP-4を埋植したときに形成される異所性骨の形態、蛋白組成に類似するものである。実際、仮骨形成時においてはその初期にBMP-4遺伝子の多量合成がin situハイブリダイゼーション、ノーザンハイブリダイゼーション法を用いて明らかにされた。BMP-4遺伝子が活性化される胎児骨形成期においてもオステオポンチン、オステオネクチンが多量に合成されることより、膜性骨化、内軟骨性骨化の如何に関わらず、未分化間葉系細胞はまず、BMP-4を合成し、増殖と分化の過程を経た後にオステオポンチン、オステオネクチンという骨基質蛋白の合成を起こして仮骨組識を形成していく共通の反応系が分子生物学的な手法を用いて明らかにされた。さらにこの過程が病理学的な石灰化においても再現されるかという問題についても検討を加え、ノーザンハイブリダイゼーション、in situハイブリダイゼーション法を用いて粥状動脈硬化層、乳癌組織、腎臓結石形成時におけるオステオポンチン、オステオネクチン遺伝子mRNA合成を検討したところ、特にオステオポンチン遺伝子を発現する細胞が石灰化に深く関わっていることが明らかとなった。しかし、骨組織における場合と異なり、粥状動脈硬化層、乳癌組織においてオステオポンチン遺伝子を発現する細胞はマクロファージであり、腎臓結石形成の場合は遠位腎尿細管上皮細胞であった。このことは、全ての石灰化になんらかの共通機構が存在することを示しており、わけてもオステオポンチンの遺伝子発現をもってこれを予想することが可能であることを示している。

研究成果

(34件)

すべてその他

すべて文献書誌 (34件)

[文献書誌] E,Kondo: "Detection of bcl-2 messenger RNA in normal and neoplastic lymphoid tissues by immunohistochemical and in situ hybridization techniques." Blood. 80. 2044-2051 (1992)
[文献書誌] D,R,Edwards: "Expression of TIMP in fetal and adult mouse tissues studied by in situ hybridization." Matrix,Suppl.1. 286-293 (1992)
[文献書誌] T,Tono: "c-kit gene was not transcribed in cultured mast cells of mast cell deficient W^<sh>/W^<sh> mice that have normal number of erythrocyte and normal c-kit coding region." Blood. 80. 1448-1453 (1992)
[文献書誌] H,Onoue: "Absence of immature mast cells in the skin of Ws/Ws rats with a small deletion at tyrosine kinase domain of the c-kit gene." Ann.J.Pathol.142. 1001-1007 (1993)
[文献書誌] S,Nomura: "Method for detecting the expression of bone matrix proteins by in situ hybridization using decalcified mineralized tissue." Acta.Histochem.Cytochem. 26. 299-305 (1993)
[文献書誌] K,Kohri: "Structure and Expression of the mRNA encoding urinary stone protein(osteopontin)." J.Biol Chem.268. 15180-15184 (1993)
[文献書誌] S,Osada: "Predominant expression of nPKCη,a Ca2+ -independent isoform of protein kinase C in Epithelial tissue,in association with epithelial differentiation." Cell,Growth and Differentiation. 4. 167-175 (1993)
[文献書誌] S,Hirota: "Expression of osteopontin mRNA by macrophages in atherosclerotic plaques:A possible association with calcification" Ann.J.Pathol.143. 1003-1008 (1993)
[文献書誌] E,Morii: "Types of neurons that express mRNA of c-kit and its ligand in developing murine cerebellum." Acta Histochem,Cytochem. 26. 359-364 (1993)
[文献書誌] J,Nagoshi: "Expression of bone related genes by spindle-shaped and chondrocyte-like cells that appeared in androgen-dependent SC115 tumors after androgen removal." Lal,Invest.(in press).
[文献書誌] T,Nakase: "Expression of bone morphogenetic protein-4 mRNA in the initial stage of fracture healing." J.of Bone and Mineral Res. 9. 651-659 (1994)
[文献書誌] U,Koshimizu: "Wn mutation of c-kit receptor affects its post-translational processing and extracellular expression." Orcogene. 8. 157-162 (1994)
[文献書誌] S,Hirota: "Possible involvement of c-kit receptor and its ligand in increase of mast cells in neurofibroma tissues." Arch.Pathol Lal.Med. 117. 996-999 (1993)
[文献書誌] N.Takemura: "Expression and localization of oxitocin receptor gene in human uterine endometrium in relation to the menstrual cycle" Endocrinology. 132. 1830-1835 (1993)
[文献書誌] T.Tsujimura: "Mast cell number in the skin of heterozygote reflects the molecular nature of c-kit mutation." Blood. 81. 2530-2538 (1993)
[文献書誌] T,Kasugai: "Deficient differentiation of mast cells in the skin of mi/mi mice:Usefulness of in situ hybridization for evaluation of mast cell phenotype." Ann.J.Pathol. 143. 1337-1347 (1993)
[文献書誌] T,Hirata: "Stem cell factor induces outgrowth of c-kit-positive neurites from dorsal root ganglia of mouse embryos." Development. 119. 49-56 (1993)
[文献書誌] H,Saeki: "Detection of low copy number of Epstein-Barr virus by in situ hybridization by non-radioisotopic probes prepared by polymerase chain reaction." Diagnotic Molecular Pathology. 2. 108-115 (1993)
[文献書誌] K,Hirakawa: "Expression and localization of the mRNA for bone matrix proteins during fracture healing revealed as in situ hybridization." J.of Bone and Mineral Res. (印刷中).
[文献書誌] Y,Nakajima: "Identification of a cell types of odontoma-like clusters in microphtalmic(mi/mi)mice using in situ hybridization." J.of Dental Res.(印刷中).
[文献書誌] S,Hirota: "Expression of bone related genes in ectopically developing bone by non-radioisotopic in situ hybridization." Cell and Tissue Res.(印刷中).
[文献書誌] T.Takemura: "Localization of osteopontin in the otolithic organ of adult rats." Hearing Res. (印刷中).
[文献書誌] E.Morii: "Demonstration of retinal cells expressing messenger RNAs of the c-kit receptor and its ligand." Neuroscience Lett.(印刷中).
[文献書誌] T,Yoshino: "Inverse expression of bcl-2 protein and Fas antigen in lymphoblasts in peripheral lymph nodes and activated blood T and B lymphocytes." Blood. (印刷中).
[文献書誌] T,Kuroki: "Protein kinase C in skin:its activation,down regulation,substrate proteins and family members." Excerpta Medica. (印刷中).
[文献書誌] T,Tsujimura: "Ligand-independent activation of c-kit receptor tyrosine kinase in a murine mastocytoma cell linr P-815 generated by a point mutation." Blood. (印刷中).
[文献書誌] A,Ito: "Developmental expression of Flt3 mRNA in the mouse brain." J.of Mol.Neuroscience. (印刷中).
[文献書誌] N.Huh: "Isolation and characterization of a novel hair follicle-specific gene,hac-1." J.Invest,Dermatology. (印刷中).
[文献書誌] H.Kim: "PKCγ gene expression is delayed in postnatal central nervous system of mi/mi mice." J.of Molecular Neuroscience. (印刷中).
[文献書誌] Y,Kaisho: "Expression of neurotrophin genes in the brain of senescence-accerated mouse (SAM) during postnatal development." Brain Res. (印刷中).
[文献書誌] M,Takemura: "Expression and localization of human oxytocin receptor mRNA and its protein in chcrion and decidua during parturition." J.Clin.Invest. (印刷中).
[文献書誌] M.Yamazaki: "c-kit gene is expressed by skin mast cells in embryos but not in puppies of W^<sh>/W^<sh> mice:Age dependent abolishment of c-kit gene expression." Blood. (印刷中).
[文献書誌] Y,Kitamura: "In immunopharmacology of mast cells and basophils." Academic Press.London, 232 (1993)
[文献書誌] Y,Kitamura: "Growth fuctors in Cell growth morphogenesis" Gann Monograph in Cancer research,inc(印刷中), (1994)

1993 年度 実績報告書

骨折治癒の分子病理学的解析

研究代表者

野村 慎太郎 大阪大学, 医学部, 助教授 (80159087)

研究成果

[文献書誌] E,Kondo: "Detection of bcl-2 messenger RNA in normal and neoplastic lymphoid tissues by immunohistochemical and in situ hybridization techniques." Blood. 80. 2044-2051 (1992)

[文献書誌] D,R,Edwards: "Expression of TIMP in fetal and adult mouse tissues studied by in situ hybridization." Matrix,Suppl.1. 286-293 (1992)

[文献書誌] T,Tono: "c-kit gene was not transcribed in cultured mast cells of mast cell deficient W^<sh>/W^<sh> mice that have normal number of erythrocyte and normal c-kit coding region." Blood. 80. 1448-1453 (1992)

[文献書誌] H,Onoue: "Absence of immature mast cells in the skin of Ws/Ws rats with a small deletion at tyrosine kinase domain of the c-kit gene." Ann.J.Pathol.142. 1001-1007 (1993)

[文献書誌] S,Nomura: "Method for detecting the expression of bone matrix proteins by in situ hybridization using decalcified mineralized tissue." Acta.Histochem.Cytochem. 26. 299-305 (1993)

[文献書誌] K,Kohri: "Structure and Expression of the mRNA encoding urinary stone protein(osteopontin)." J.Biol Chem.268. 15180-15184 (1993)

[文献書誌] S,Osada: "Predominant expression of nPKCη,a Ca2+ -independent isoform of protein kinase C in Epithelial tissue,in association with epithelial differentiation." Cell,Growth and Differentiation. 4. 167-175 (1993)

[文献書誌] S,Hirota: "Expression of osteopontin mRNA by macrophages in atherosclerotic plaques:A possible association with calcification" Ann.J.Pathol.143. 1003-1008 (1993)

[文献書誌] E,Morii: "Types of neurons that express mRNA of c-kit and its ligand in developing murine cerebellum." Acta Histochem,Cytochem. 26. 359-364 (1993)

[文献書誌] J,Nagoshi: "Expression of bone related genes by spindle-shaped and chondrocyte-like cells that appeared in androgen-dependent SC115 tumors after androgen removal." Lal,Invest.(in press).

[文献書誌] T,Nakase: "Expression of bone morphogenetic protein-4 mRNA in the initial stage of fracture healing." J.of Bone and Mineral Res. 9. 651-659 (1994)

[文献書誌] U,Koshimizu: "Wn mutation of c-kit receptor affects its post-translational processing and extracellular expression." Orcogene. 8. 157-162 (1994)

[文献書誌] S,Hirota: "Possible involvement of c-kit receptor and its ligand in increase of mast cells in neurofibroma tissues." Arch.Pathol Lal.Med. 117. 996-999 (1993)

[文献書誌] N.Takemura: "Expression and localization of oxitocin receptor gene in human uterine endometrium in relation to the menstrual cycle" Endocrinology. 132. 1830-1835 (1993)

[文献書誌] T.Tsujimura: "Mast cell number in the skin of heterozygote reflects the molecular nature of c-kit mutation." Blood. 81. 2530-2538 (1993)

[文献書誌] T,Kasugai: "Deficient differentiation of mast cells in the skin of mi/mi mice:Usefulness of in situ hybridization for evaluation of mast cell phenotype." Ann.J.Pathol. 143. 1337-1347 (1993)

[文献書誌] T,Hirata: "Stem cell factor induces outgrowth of c-kit-positive neurites from dorsal root ganglia of mouse embryos." Development. 119. 49-56 (1993)

[文献書誌] H,Saeki: "Detection of low copy number of Epstein-Barr virus by in situ hybridization by non-radioisotopic probes prepared by polymerase chain reaction." Diagnotic Molecular Pathology. 2. 108-115 (1993)

[文献書誌] K,Hirakawa: "Expression and localization of the mRNA for bone matrix proteins during fracture healing revealed as in situ hybridization." J.of Bone and Mineral Res. (印刷中).

[文献書誌] Y,Nakajima: "Identification of a cell types of odontoma-like clusters in microphtalmic(mi/mi)mice using in situ hybridization." J.of Dental Res.(印刷中).

[文献書誌] S,Hirota: "Expression of bone related genes in ectopically developing bone by non-radioisotopic in situ hybridization." Cell and Tissue Res.(印刷中).

[文献書誌] T.Takemura: "Localization of osteopontin in the otolithic organ of adult rats." Hearing Res. (印刷中).

[文献書誌] E.Morii: "Demonstration of retinal cells expressing messenger RNAs of the c-kit receptor and its ligand." Neuroscience Lett.(印刷中).

[文献書誌] T,Yoshino: "Inverse expression of bcl-2 protein and Fas antigen in lymphoblasts in peripheral lymph nodes and activated blood T and B lymphocytes." Blood. (印刷中).

[文献書誌] T,Kuroki: "Protein kinase C in skin:its activation,down regulation,substrate proteins and family members." Excerpta Medica. (印刷中).

[文献書誌] T,Tsujimura: "Ligand-independent activation of c-kit receptor tyrosine kinase in a murine mastocytoma cell linr P-815 generated by a point mutation." Blood. (印刷中).

[文献書誌] A,Ito: "Developmental expression of Flt3 mRNA in the mouse brain." J.of Mol.Neuroscience. (印刷中).

[文献書誌] N.Huh: "Isolation and characterization of a novel hair follicle-specific gene,hac-1." J.Invest,Dermatology. (印刷中).

[文献書誌] H.Kim: "PKCγ gene expression is delayed in postnatal central nervous system of mi/mi mice." J.of Molecular Neuroscience. (印刷中).

[文献書誌] Y,Kaisho: "Expression of neurotrophin genes in the brain of senescence-accerated mouse (SAM) during postnatal development." Brain Res. (印刷中).

[文献書誌] M,Takemura: "Expression and localization of human oxytocin receptor mRNA and its protein in chcrion and decidua during parturition." J.Clin.Invest. (印刷中).

[文献書誌] M.Yamazaki: "c-kit gene is expressed by skin mast cells in embryos but not in puppies of W^<sh>/W^<sh> mice:Age dependent abolishment of c-kit gene expression." Blood. (印刷中).

[文献書誌] Y,Kitamura: "In immunopharmacology of mast cells and basophils." Academic Press.London, 232 (1993)

[文献書誌] Y,Kitamura: "Growth fuctors in Cell growth morphogenesis" Gann Monograph in Cancer research,inc(印刷中), (1994)

1993 年度実績報告書

野村慎太郎大阪大学, 医学部, 助教授 (80159087)