2002 Fiscal Year Annual Research Report

in situ ノックアウト法の確立

Research Project

Project/Area Number	13557002
Research Institution	Shimane Medical University
Principal Investigator	大谷浩島根医科大学, 医学部, 教授 (20160533)
Co-Investigator(Kenkyū-buntansha)	宇田川潤島根医科大学, 医学部, 助手 (10284027) 橋本龍樹島根医科大学, 医学部, 助手 (90252907) 八田稔久島根医科大学, 医学部, 助教授 (20238025)
Keywords	マウス胎児 / 子宮外発生法 / specific antagonist / enzyme inhibitor
Research Abstract	昨年度に引き続き、胎児操作法の改良ならびに特異的antagonist等の導入に関する基本的実験条件の確立と抑制効率の上昇のための実験、神経内分泌系の組織形成における特異的antagonistの効果についての予備的実験を行った。 (1)胎児操作・導入技術の改善に関する実験。マウス妊娠9および10日に羊水中へGFP遺伝子ベクターを注入して皮膚へ遺伝子導入をはかる実験系につき、さらに長期的な安定性へ向けた条件の検討を行った。妊娠10日に2cfu/μlを2μl注入する系で、生後も継続して遺伝子発現を観察することができた。また特異的antagonistの注入によるノックアウト効果をより長時間持続させるため、抑制物質を吸着させたビーズを注入する方法について、当教室で報告している耳胞における細胞死に対して、capsase inhibitorによる細胞死抑制、staurosporine (Kinase inhibitor)を用いて検討した。ビーズを注入すること自体は可能であることが、注入後の組織学的観察から確認されたが、注入可能なビーズ径が小さいためか、ビーズ使用による明らかな延長効果は現在のところ認められておらず、さらに検討が必要である。 (2)神経内分泌系の形成に関わる分子、特にNPYおよびその特異的antagonist (NPY-A)の注入実験。胎児期への導入の予備実験として新生児期においてNPYとNPY-Aの投与による効果を検討・確認した。プレプロ甲状腺刺激放出ホルモンの発現に対する両者の効果の違いをRNA blot, RT-PCR, in situ hybridizationにより観察した。現在、この系をさらに胎児期に用いて解析中である。

Research Products
(8 results)

All Other

All Publications (8 results)

[Publications] Richard Cousley: "Validity of the Hfm transgenic mouse as a model for hemifacial microsomia"Cleft Palate Craniofacial J.. 39. 81-92 (2002)
[Publications] Akihiro Matsumoto: "Sequential observation of exencephaly and subsequent morphological changes by mouse exo utero development system : analysis of the mechanism of transformation from exencephaly to anencephaly"Anat. Embryol.. 205. 7-18 (2002)
[Publications] Akihiro Matsumoto: "Occlusion and subsequent re-canalization in early duodenal development in human embryos : integrated organogenesis and histogenesis through a possible epithelial-mesenchymal interaction epithelial-mesenchymal interaction"Anat. Embryol.. 205. 53-65 (2002)
[Publications] Yukiko Kagohashi: "PSK, a biological response modifier, modifies p53 expression, mitosis and apoptosis in X-ray irradiated mouse embryos : Possible cellular mechanism of the anti-teratogenic effect"Congenital Anomalies. 42. 21-26 (2002)
[Publications] Toshihisa Hatta: "The role of gp130 in cerebral cortical development : in vivo functional analysis in a mouse exo utero system"J. Neurosci.. 22. 5516-5524 (2002)
[Publications] Ryuju Hashimoto: "Perinatal development of the rat hip joint with restrained fetal movement"Congenital Anomalies. 42. 21-26 (2002)
[Publications] Toshihisa Hatta: "Opposite effects of the maternal immune system activated by interleukin-lb vs. PSK and OK432 on 5-azacytidine-induced birth defects"Cong. Anom.. 43(In press). (2003)
[Publications] Yukiko Kagohashi: "PSK, a biological response modifier, induces a systemic developmental delay in X-ray irradiated mouse embryos : coincidence with its anti-teratogenic effect"Shimane J. Med. Sci.. (In press). (2003)

2002 Fiscal Year Annual Research Report

in situ ノックアウト法の確立

Principal Investigator

大谷 浩 島根医科大学, 医学部, 教授 (20160533)

Research Products

[Publications] Richard Cousley: "Validity of the Hfm transgenic mouse as a model for hemifacial microsomia"Cleft Palate Craniofacial J.. 39. 81-92 (2002)

[Publications] Akihiro Matsumoto: "Sequential observation of exencephaly and subsequent morphological changes by mouse exo utero development system : analysis of the mechanism of transformation from exencephaly to anencephaly"Anat. Embryol.. 205. 7-18 (2002)

[Publications] Akihiro Matsumoto: "Occlusion and subsequent re-canalization in early duodenal development in human embryos : integrated organogenesis and histogenesis through a possible epithelial-mesenchymal interaction epithelial-mesenchymal interaction"Anat. Embryol.. 205. 53-65 (2002)

[Publications] Yukiko Kagohashi: "PSK, a biological response modifier, modifies p53 expression, mitosis and apoptosis in X-ray irradiated mouse embryos : Possible cellular mechanism of the anti-teratogenic effect"Congenital Anomalies. 42. 21-26 (2002)

[Publications] Toshihisa Hatta: "The role of gp130 in cerebral cortical development : in vivo functional analysis in a mouse exo utero system"J. Neurosci.. 22. 5516-5524 (2002)

[Publications] Ryuju Hashimoto: "Perinatal development of the rat hip joint with restrained fetal movement"Congenital Anomalies. 42. 21-26 (2002)

[Publications] Toshihisa Hatta: "Opposite effects of the maternal immune system activated by interleukin-lb vs. PSK and OK432 on 5-azacytidine-induced birth defects"Cong. Anom.. 43(In press). (2003)

[Publications] Yukiko Kagohashi: "PSK, a biological response modifier, induces a systemic developmental delay in X-ray irradiated mouse embryos : coincidence with its anti-teratogenic effect"Shimane J. Med. Sci.. (In press). (2003)

大谷浩島根医科大学, 医学部, 教授 (20160533)