1999 Fiscal Year Annual Research Report

海馬及び運動ニューロン生存を担う新規神経栄養因子の探索とその機能解析

Research Project

Project/Area Number	11157217
Research Institution	Osaka University
Principal Investigator	船越洋大阪大学, 医学系研究科, 助手 (40273685)
Co-Investigator(Kenkyū-buntansha)	中村敏一大阪大学, 医学系研究科, 教授 (00049397)
Keywords	肝細胞増殖因子 / HGF / HLP / Gas6 / 神経栄養因子 / ニューロン / クローニング / トランスジェニックマウス
Research Abstract	(1)海馬ニューロンに対するHGFおよび新規神経栄養因子Gas6のもつin vitro神経栄養活性の解析 HGFは新規神経栄養因子であるが、これまで明かとなっていた海馬ニューロンの血清除去による神経細胞死モデルに加え、グルタミン酸誘導性神経細胞死モデルやアミロイドβ25-35誘導性神経細胞死をHGFが濃度依存性に阻止することを明らかにした。また脳神経系特異的チロシンキナーゼ型受容体SkyのリガンドGas6が新規神経栄養因子として機能し、海馬、さらには小脳顆粒細胞に対し神経生存促進活性をもつことを同定した。 (2)脳神経系特異的チロシンキナーゼ型受容体Skyのファミリー分子のクローニングとその機能解析新規神経栄養因子として昨年同定したGas6の受容体Skyにはファミリー受容体Axlおよびc-Merが知られる。私達はアフリカズメガエルから第4番目のファミリー受容体をクローニングしXksyと名付けた。このリガンドはまだ不明であるためXksyの膜貫通および細胞内領域とEGFの細胞外領域とのキメラタンパクを作成した。現在この分子をEGFで活性化しXksyの機能を解析している。 (3)HGFファミリー分子(HLP)の新規神経栄養因子としての同定 HGF-like protein(HLP)はHGFと同様4つのクリングル構造をもつアルファ鎖とセリンプロテアーゼ様構造をもつベータ鎖よりなる。これまでHLPはマクロファージに対する機能しか知られていなかったが、本研究でHLPが鶏胚感覚神経節にin vitroで神経栄養活性(神経突起伸長・migration促進活性)を示すことを明かとした。HLPの活性の特異性は、Ron/HLPレセプターの細胞外ドメインとイムノグロブリンFcとのキメラタンパクにより確認された。 (4)運動神経変性疾患モデルマウスに対する新規神経栄養因子(HGF)の機能解析 (a)神経系特異的HGF発現トランスジェニックマウス(NE-HGF-Tg)の作成生後活性化される神経特異的プロモーター制御下にHGF-tagを発現する神経特異的HGF発現マウス(NS-Tg-HGF)を作成した。 (b)NS-HGF-Tgと筋萎縮性側索硬化症(ALS)モデルTg-マウスであるSOD-1変異遺伝子Tg-マウスの交配によるダブルTg-マウスの作成とHGFのALSにおける機能解析ダブルTg-マウスではHGF遺伝子を脳血液関門を考慮せず長期間ALS-Tg-マウスの神経に直接発現できる。ダブルTg-マウスでALSの発症遅延と生存期間の解析中である。

Research Products
(37 results)

All Other

All Publications (37 results)

[Publications] W. Sun, H. Funakoshi, K. Matsumoto and T. Nakamura: "A sensitive quantification method for evaluating the level of HGF and c-met/HGF receptor mRNAs in the nervous system using competitive RT-PCR"Brain Res. Protocol. (In press). (2000)
[Publications] W. Sun, H. Funakoshi and T. Nakamura: "Differential expression of hepatocyte growth factor and its receptor, c-Met in the rat retina during development"Brain Res.. 851. 46-53 (1999)
[Publications] K. Kosai, K. Matsumoto, H. Funakoshi and T. Nakamura: "Hepatocyte growth factor prevents endotoxin-induced lethal hepatic failure in mice"Hepatology. 30. 151-159 (1999)
[Publications] J. Wojta, et al.: "Hepatocyte growth factor increases expression of vascular endothelial growth factor and plasminogen activator inhibitor-1 in human keratinocytes and the vascular endothelial growth factor receptor flk-1 in human endothelial cells Evidence for a paracrine loop in wound healing"Lab. Invest. 79. 427-438 (1999)
[Publications] Z. Jiao, et al.: "Effect of D-galactosamine hydrochloride and partial hepatectomy on spontaneous hepatic injury and hepatocarcinogenesis in Long Evans Cinamon rats"Jpn. J. Cancer Res.. 90. 496-504 (1999)
[Publications] H. Azuma, et al.: "Effect of hepatocyte growth factor on chronic rejection in rat renal allografts"Transplat. Proc.. 31. 854-855 (1999)
[Publications] S. Takada, et al.: "Effect of hepatocyte growth factor on tacrolimus-induced nephrotoxicity in spontaneously hypertensive rats"Transpl. Int.. 12. 27-32 (1999)
[Publications] H. Ueda, et al.: "Gene transfection of hepatocyte growth factor attenuates reperfusion injury in the heart"Annals. Thoracic Surg.. 67. 1726-1731 (1999)
[Publications] K. Kosai, K. Matsumoto, and T. Nakamura: "Hepatocyte growth factor prevents endotoxin-induced lethal hepatic failure in mice"Hepatology. 30. 151-159 (1999)
[Publications] R. Hasina, et al.: "Autocrine and paracrine motility factors and their involvement in invasiveness in a human oral carcinoma cell line"Brit. J. Cancer. 80. 1708-1717 (1999)
[Publications] K. Kitajima, U. Koshimizu and T. Nakamura: "Expression of a novel type of classic cadherin, PB-cadherin in developing brain and limb buds"Develop. Dyn.. 215. 206-214 (1999)
[Publications] N. Taniura-Matsumoto, K. Matsumoto and T. Nakamura: "Prostaglandin production in mouse mammary carcinoma cells confers invasive growth potential by inducing hepatocyte growth factor in fibroblasts"Brit. J. Cancer. 81. 194-202 (1999)
[Publications] M. Kato, Y. Kato, T. Nakamura and Y. Sugiyama: "Efficient extraction by the liver governs the overall elimination of hepatocyte growth factor in rats"J. Pharmacol. Exp. Therap.. 290. 373-379 (1999)
[Publications] Morishita, et al.: "Therapeutic angiogenesis induced by human recombinant hepatocyte growth factor in rabbit hind limb ischemia model as a cytokine supplement therapy"Hypertention. 33. 1379-1384 (1999)
[Publications] W. G. Jiang, et al.: "Hepatocyte growth factor/scatter factor decreases the expression of occuludin and transendothelial resistance (TER) and increased paracellular permeability in human vascular endothelial cells"J. Cell. Physiol.. 181. 319-329 (1999)
[Publications] C. C. Fehlner-Gardiner, et al.: "Characterization of a functional relationship between hepatocyte growth factor and mouse bone marrow-derived mast cells"Differentiation. 65. 27-42 (1999)
[Publications] S. Deleu, et al.: "Immediate early gene expression in dog thyrocytes in response to growth proliferation and differentiation stimuli"J. Cell. Physiol.. 181. 342-354 (1999)
[Publications] S. Deleu, et al.: "IFG-1 or insulin, and TSH cyclic AMP cascade separately control dog and human thyroid cell growth and DNA synthesis, and complement each other in inducing mitogenesis"Mole. Cell. Endocrin.. 149. 41-51 (1999)
[Publications] S. Hayashi, et al.: "Potential role of hepatocyte growth factor, a novel angiogenic growth factor in peripheral arterial disease : Downregulation of HGF in response to hypoxia in vascular cell"Circulation. 100. II-301-II-308 (1999)
[Publications] T. Nagahori, et al.: "Interferon-gamma upregulates the c-Met/HGF receptor expression in alvedar epithelial cells"Am. J. Respir. Cell Mol. Biol.. 21. 490-497 (1999)
[Publications] O. Amano, U. Koshimizu, T. Nakamura and S. Iseki: "Enhancement by hepatocyte growth factor (HGF) of bone and cartilage formation during embryonic mouse mandibular development in vitro"Arch. Oral Biol. 44. 935-946 (1999)
[Publications] K. Nishimura, et al.: "Prostate stromal cell-derived hepatocyte growth factor induces invasion of prostate cancer cells DU 145 through Tumor-stromal interaction"Prostate. 41. 145-153 (1999)
[Publications] T. Ueki, et al.: "Hepatocyte growth factor gene therapy of liver cirrhosis in rats"Nature Medicine. 5. 226-230 (1999)
[Publications] A. Uchiyama, et al.: "Hepatocyte growth factor and invasion-stimulatory activity are induced in pleural fluid by surgery in lung cancer patients"Br. J. Cancer. 81. 721-726 (1999)
[Publications] K. Matsumoto, et al.: "Prevention of renal damage by Angiotensin II blockade, accompanied by increased renal hepatocyte growth factor in experimental hypertensive rats"Hypertension. 34. 279-284 (1999)
[Publications] Y. Okura, et al.: "Analysis of neurotrophic factor of hepatocyte growth factor (HGF) in the adult hypoglossal nerve axotomy model"Eur. J. Neurosci.. 11. 4139-4144 (1999)
[Publications] T. Sumi, K. Matsumoto, Y. Takai and T. Nakamura: "Cofilin phosphorylation and actin cytoskeletal dynamics regulated by Rho- and Cdc42-activated LIM -kinase2"J. Cell Biol.. 147. 1519-1532 (1999)
[Publications] M. K. Jones, et al.: "HGF triggers activation of Cox-2 gene in rat gastric epithelial cells Action mediated through the ERK2 signalling pathway"FASEB J.. 13. 2186-2194 (1999)
[Publications] Y. Kawano, et al.: "Phosphorylation of myosin-binding subunit (MBS) of myosin phosphatase by Rho-kinase in vivo"J. Cell Biol.. 147. 1023-1037 (1999)
[Publications] F. Tamura, et al.: "FK506 promotes liver regeneration by suppressing natural killer cell activity"J. Gastroenterol. and Hepatol.. (in press). (2000)
[Publications] A. Uchiyama, et al.: "Significance of locally-secreted hepatocyte growth factor after thoracic cancer surgery as a stimulator for postoperative cancer invasion"Jpn. J. Clin. Oncol.. (in press). (2000)
[Publications] W. G. Jiang, et al.: "Antagonistic effect of NK4, a novel HGF variant, on in vitro angiogenesis of human vascular endothelial cells"Clin. Cancer Res.. (in press). (2000)
[Publications] S. Hiscox, et al.: "Inhibition of HGF/SF-induced breast cancer cell matility and invasion by the HGF/SF variant, NK4"Breast Cancer Res. & Treat.. (in press). (2000)
[Publications] M. nakano, et al.: "Hepatocyte growth factor is essential for amelioration of hyperglycemia in strepto zotocin-induced diabetic mice receiving a marginal mass of intrahepatic islet grafts"Transplantation. (in press). (2000)
[Publications] T. Yamada, et al.: "Enhanced expression of hepatocyte growth factor by pulmonary ischemia reperfusion injury in the rat"Am. J. Resp. Crit Care Med.. (in press). (2000)
[Publications] 船越洋、中村敏一: "肝細胞増殖因子因子(HGF)の検査値と各種疾患"日本臨床. 57. 821-826 (1999)
[Publications] 船越洋、紀氏 A.優子、中村敏一: "キメラタンパク作成法と利用"脳の科学. 21. 537-542 (1999)

1999 Fiscal Year Annual Research Report

海馬及び運動ニューロン生存を担う新規神経栄養因子の探索とその機能解析

Principal Investigator

船越 洋 大阪大学, 医学系研究科, 助手 (40273685)

Research Products

[Publications] W. Sun, H. Funakoshi, K. Matsumoto and T. Nakamura: "A sensitive quantification method for evaluating the level of HGF and c-met/HGF receptor mRNAs in the nervous system using competitive RT-PCR"Brain Res. Protocol. (In press). (2000)

[Publications] W. Sun, H. Funakoshi and T. Nakamura: "Differential expression of hepatocyte growth factor and its receptor, c-Met in the rat retina during development"Brain Res.. 851. 46-53 (1999)

[Publications] K. Kosai, K. Matsumoto, H. Funakoshi and T. Nakamura: "Hepatocyte growth factor prevents endotoxin-induced lethal hepatic failure in mice"Hepatology. 30. 151-159 (1999)

[Publications] Z. Jiao, et al.: "Effect of D-galactosamine hydrochloride and partial hepatectomy on spontaneous hepatic injury and hepatocarcinogenesis in Long Evans Cinamon rats"Jpn. J. Cancer Res.. 90. 496-504 (1999)

[Publications] H. Azuma, et al.: "Effect of hepatocyte growth factor on chronic rejection in rat renal allografts"Transplat. Proc.. 31. 854-855 (1999)

[Publications] S. Takada, et al.: "Effect of hepatocyte growth factor on tacrolimus-induced nephrotoxicity in spontaneously hypertensive rats"Transpl. Int.. 12. 27-32 (1999)

[Publications] H. Ueda, et al.: "Gene transfection of hepatocyte growth factor attenuates reperfusion injury in the heart"Annals. Thoracic Surg.. 67. 1726-1731 (1999)

[Publications] K. Kosai, K. Matsumoto, and T. Nakamura: "Hepatocyte growth factor prevents endotoxin-induced lethal hepatic failure in mice"Hepatology. 30. 151-159 (1999)

[Publications] R. Hasina, et al.: "Autocrine and paracrine motility factors and their involvement in invasiveness in a human oral carcinoma cell line"Brit. J. Cancer. 80. 1708-1717 (1999)

[Publications] K. Kitajima, U. Koshimizu and T. Nakamura: "Expression of a novel type of classic cadherin, PB-cadherin in developing brain and limb buds"Develop. Dyn.. 215. 206-214 (1999)

[Publications] N. Taniura-Matsumoto, K. Matsumoto and T. Nakamura: "Prostaglandin production in mouse mammary carcinoma cells confers invasive growth potential by inducing hepatocyte growth factor in fibroblasts"Brit. J. Cancer. 81. 194-202 (1999)

[Publications] M. Kato, Y. Kato, T. Nakamura and Y. Sugiyama: "Efficient extraction by the liver governs the overall elimination of hepatocyte growth factor in rats"J. Pharmacol. Exp. Therap.. 290. 373-379 (1999)

[Publications] Morishita, et al.: "Therapeutic angiogenesis induced by human recombinant hepatocyte growth factor in rabbit hind limb ischemia model as a cytokine supplement therapy"Hypertention. 33. 1379-1384 (1999)

[Publications] W. G. Jiang, et al.: "Hepatocyte growth factor/scatter factor decreases the expression of occuludin and transendothelial resistance (TER) and increased paracellular permeability in human vascular endothelial cells"J. Cell. Physiol.. 181. 319-329 (1999)

[Publications] C. C. Fehlner-Gardiner, et al.: "Characterization of a functional relationship between hepatocyte growth factor and mouse bone marrow-derived mast cells"Differentiation. 65. 27-42 (1999)

[Publications] S. Deleu, et al.: "Immediate early gene expression in dog thyrocytes in response to growth proliferation and differentiation stimuli"J. Cell. Physiol.. 181. 342-354 (1999)

[Publications] S. Deleu, et al.: "IFG-1 or insulin, and TSH cyclic AMP cascade separately control dog and human thyroid cell growth and DNA synthesis, and complement each other in inducing mitogenesis"Mole. Cell. Endocrin.. 149. 41-51 (1999)

[Publications] S. Hayashi, et al.: "Potential role of hepatocyte growth factor, a novel angiogenic growth factor in peripheral arterial disease : Downregulation of HGF in response to hypoxia in vascular cell"Circulation. 100. II-301-II-308 (1999)

[Publications] T. Nagahori, et al.: "Interferon-gamma upregulates the c-Met/HGF receptor expression in alvedar epithelial cells"Am. J. Respir. Cell Mol. Biol.. 21. 490-497 (1999)

[Publications] O. Amano, U. Koshimizu, T. Nakamura and S. Iseki: "Enhancement by hepatocyte growth factor (HGF) of bone and cartilage formation during embryonic mouse mandibular development in vitro"Arch. Oral Biol. 44. 935-946 (1999)

[Publications] K. Nishimura, et al.: "Prostate stromal cell-derived hepatocyte growth factor induces invasion of prostate cancer cells DU 145 through Tumor-stromal interaction"Prostate. 41. 145-153 (1999)

[Publications] T. Ueki, et al.: "Hepatocyte growth factor gene therapy of liver cirrhosis in rats"Nature Medicine. 5. 226-230 (1999)

[Publications] A. Uchiyama, et al.: "Hepatocyte growth factor and invasion-stimulatory activity are induced in pleural fluid by surgery in lung cancer patients"Br. J. Cancer. 81. 721-726 (1999)

[Publications] K. Matsumoto, et al.: "Prevention of renal damage by Angiotensin II blockade, accompanied by increased renal hepatocyte growth factor in experimental hypertensive rats"Hypertension. 34. 279-284 (1999)

[Publications] Y. Okura, et al.: "Analysis of neurotrophic factor of hepatocyte growth factor (HGF) in the adult hypoglossal nerve axotomy model"Eur. J. Neurosci.. 11. 4139-4144 (1999)

[Publications] T. Sumi, K. Matsumoto, Y. Takai and T. Nakamura: "Cofilin phosphorylation and actin cytoskeletal dynamics regulated by Rho- and Cdc42-activated LIM -kinase2"J. Cell Biol.. 147. 1519-1532 (1999)

[Publications] M. K. Jones, et al.: "HGF triggers activation of Cox-2 gene in rat gastric epithelial cells Action mediated through the ERK2 signalling pathway"FASEB J.. 13. 2186-2194 (1999)

[Publications] Y. Kawano, et al.: "Phosphorylation of myosin-binding subunit (MBS) of myosin phosphatase by Rho-kinase in vivo"J. Cell Biol.. 147. 1023-1037 (1999)

[Publications] F. Tamura, et al.: "FK506 promotes liver regeneration by suppressing natural killer cell activity"J. Gastroenterol. and Hepatol.. (in press). (2000)

[Publications] A. Uchiyama, et al.: "Significance of locally-secreted hepatocyte growth factor after thoracic cancer surgery as a stimulator for postoperative cancer invasion"Jpn. J. Clin. Oncol.. (in press). (2000)

[Publications] W. G. Jiang, et al.: "Antagonistic effect of NK4, a novel HGF variant, on in vitro angiogenesis of human vascular endothelial cells"Clin. Cancer Res.. (in press). (2000)

[Publications] S. Hiscox, et al.: "Inhibition of HGF/SF-induced breast cancer cell matility and invasion by the HGF/SF variant, NK4"Breast Cancer Res. & Treat.. (in press). (2000)

[Publications] M. nakano, et al.: "Hepatocyte growth factor is essential for amelioration of hyperglycemia in strepto zotocin-induced diabetic mice receiving a marginal mass of intrahepatic islet grafts"Transplantation. (in press). (2000)

[Publications] T. Yamada, et al.: "Enhanced expression of hepatocyte growth factor by pulmonary ischemia reperfusion injury in the rat"Am. J. Resp. Crit Care Med.. (in press). (2000)

[Publications] 船越洋、中村敏一: "肝細胞増殖因子因子(HGF)の検査値と各種疾患"日本臨床. 57. 821-826 (1999)

[Publications] 船越洋、紀氏 A.優子、中村敏一: "キメラタンパク作成法と利用"脳の科学. 21. 537-542 (1999)

船越洋大阪大学, 医学系研究科, 助手 (40273685)