1993 Fiscal Year Annual Research Report

チロシンキナーゼ受容体癌原遺伝子の発現を制御するmi遺伝子の分離同定

Research Project

Project/Area Number	05152071
Research Institution	Osaka University
Principal Investigator	野村慎太郎大阪大学, 医学部, 助教授 (80159087)
Co-Investigator(Kenkyū-buntansha)	北村幸彦大阪大学, 医学部, 教授 (70028520) 廣田誠一大阪大学, 医学部, 助手 (50218856) 辻村亨大阪大学, 医学部, 助手 (20227408) 春日井務大阪大学, 医学部, 助手 (80214310)
Keywords	チロシンキナーゼ / 細胞接着 / 骨大理石症マウス / DNA結合タンパク / サブトラクション / マスト細胞 / マウスミュータント / ★骨細胞
Research Abstract	mi遺伝子に異常があるマウスは骨大理石症、小眼症、メラノサイト欠損症等重篤な症状を示す。これらの症状のかなり多くの部分はmiミュータントマウスにおいて癌原遺伝子であるレセプターチロシンキナーゼc-kit遺伝子の発現が抑制されていることによるものであることが判明した。我々によってmi遺伝子単離が遂行されているときにmi遺伝子そのものと思われる遺伝子の構造の一部が米国の研究者らによって報告された。この情報をもとに我々はその遺伝子を単離し、その遺伝子発現部位をin situハイブリダイゼーション法を用いて検討した。その結果、c-kit発現細胞とmi遺伝子発現細胞にはかなりの共通性が認められた。そのもっともたるものが肥満細胞(マスト細胞)である。実際にmi遺伝子の欠損のあるマウスにおいてマスト細胞の数の減少が認められ、これがc-kit遺伝子の発現に重篤な影響を与えていることがほぼ明らかとなった。我々はさらにこのmi遺伝子の影響が及んでいる遺伝子群がいかなるものかを同定するためにmi遺伝子に欠損のある培養マスト細胞において低いレベルで発現をしている遺伝子を我々の開発したcDNAサブトラクション法によって分離した。それらの構造を検討した結果、分離されたcDNAクローンの一つがマスト細胞特異的なプロテアーゼP6であることが明らかとなった。この遺伝子がmi遺伝子によって直接影響を受けているのか、あるいはc-kit遺伝子産物を介しているのかについては生物学的な検討より前者であることが予想されていたが、実際、P6遺伝子の5'上流にDNA結合蛋白であるmi遺伝子の結合領域を見いだした。現在、それを生化学的な手法によって証明すると共に、mi遺伝子を特異的に認識する抗体を作成している。

Research Products

(27 results)

All Other

All Publications (27 results)

[Publications] S.Hirota: "Possible involvement of c-kit receptor and its ligand in increase of mast cells in neurofibroma tissues." Arch.Pathol Lab.Med. 117. 996-999 (1993)
[Publications] N.Takemura: "Expression and localization of oxitocin receptor gene in human uterine endometrium in relation to the menstrual cycle" Endocrinology. 132. 1830-1835 (1993)
[Publications] T.Tsujimura: "Mast cell number in the skin of heterozygote reflects the molecular nature of c-kit mutation." Blood. 81. 2530-2538 (1993)
[Publications] 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." Am.J.Pathol. 143. 1337-1347 (1993)
[Publications] T.Hirata: "Stem cell factor induces outgrowth of c-kit-positive neurites from dorsal root ganglia of mouse embryos." Development. 119. 49-56 (1993)
[Publications] 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)
[Publications] 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)
[Publications] S.Hirota: "Expression of osteopontin mRNA by macrophages in atheroscierotic plaques:A possible association with calcification" Am.J.Pathol. 143. 1003-1008 (1993)
[Publications] 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)
[Publications] J.Nagoshi: "Expression of bone related genes by spindle-shaped and chondrocyte-like cells that appeared in androgen-dependent SC115 tumors after androgen removal." Lab.Invest.(in press).
[Publications] 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)
[Publications] U.Koshimizu: "Wn mutation of c-kit receptor affects its post-translational processing and extracellular expression." Oncogene. 8. 157-162 (1994)
[Publications] 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).
[Publications] Y.Nakajima: "Identification of a cell types of odontoma-like clusters in microphthalmic(mi/mi)mice using in situ hybridization." J.of Dental Res.(印刷中).
[Publications] S.Hirota: "Expression of bone related genes in ectopically developing bone by non-radioisotopic in situ hybridization." Cell and Tissue Res.(印刷中).
[Publications] T.Takemura: "Localization of osteopontin in the otolithic organ of adult rats." Hearing Res. (印刷中).
[Publications] E.Morii: "Demonstration of retinal cells expressing messenger RNAs of the c-kit receptor and its ligand." Neuroscience Lett.(印刷中).
[Publications] 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).
[Publications] T.Kuroki: "Protein kinase C in skin:its activation,down regulation,substrate proteins and family members." Excerpta Medica. (印刷中).
[Publications] 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).
[Publications] A.Ito: "Developmental expression of Flt3 mRNA in the mouse brain." J.of Mol.Neuroscience. (印刷中).
[Publications] N.Huh: "Isolation and characterization of a novel hair follicle-specific gene,hac-1." J.Invest.Dermatology. (印刷中).
[Publications] H.Kim: "PKCγ gene expression is delayed in postnatal central nervous system of mi/mi mice." J.of Molecular Neuroscience. (印刷中).
[Publications] Y.Kaisho: "Expression of neurotrophin genes in the brain of senescence-accerated mouse(SAM)during postnatal development." Brain Res. (印刷中).
[Publications] M.Takemura: "Expression and localization of human oxytocin receptor mRNA and its protein in chorion and decidua during parturition." J.Clin.Invest. (印刷中).
[Publications] 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).
[Publications] Y.Kitamura: "In immunopharmacology of Mast cells and lasbphilis" Academic Press London, 232 (1993)

1993 Fiscal Year Annual Research Report

チロシンキナーゼ受容体癌原遺伝子の発現を制御するmi遺伝子の分離同定

Principal Investigator

野村 慎太郎 大阪大学, 医学部, 助教授 (80159087)

Research Products

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

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

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

[Publications] 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." Am.J.Pathol. 143. 1337-1347 (1993)

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

[Publications] 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)

[Publications] 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)

[Publications] S.Hirota: "Expression of osteopontin mRNA by macrophages in atheroscierotic plaques:A possible association with calcification" Am.J.Pathol. 143. 1003-1008 (1993)

[Publications] 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)

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

[Publications] 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)

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

[Publications] 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).

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

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

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

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

[Publications] 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).

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

[Publications] 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).

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

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

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

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

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

[Publications] 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).

[Publications] Y.Kitamura: "In immunopharmacology of Mast cells and lasbphilis" Academic Press London, 232 (1993)

野村慎太郎大阪大学, 医学部, 助教授 (80159087)