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Intracellular Localization and Secretion of Antioxicdant Enzymes : Developmental Profile and Pathophysiologic Implication in Children.

Research Project

Project/Area Number 08670866
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Pediatrics
Research InstitutionYamanashi Medical University

Principal Investigator

ASAYAMA Kohtaro  Yamanashi Medical University, Pediatrics, Assistant Professor, 医学部, 講師 (70129310)

Co-Investigator(Kenkyū-buntansha) KODERA Koji  Yamanashi Medical University, Pediatrics, Assistant, 医学部, 医員
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1996: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsFree radicals / Child / Development / Immunohistochemistry / Antioxidant enzymes / Gene expression / Selenium / Lipid peroxides / 免疫組織化学 / 仏ノアッセイ / 蛋白発現
Research Abstract

1. Intracellular antioxidant enzymes such as CuZn-superoxide dismutase (SOD), MnSOD and cellular glutathione peroxidase (GPX) were semi-quantitated by immunofluorescent histochemical staining-image analysis. The concentrations of the 3 enzymes increased cooperatively in epithelial tissues of various organ systems during perinatal development in rat.
2. Rocket immunoelectrophoresis of plasma GPX was developed, using specific polyclonal antibodies against the rat enzyme. This method enabled to determine the specific activity of plasma GPX in biological fluid, and was useful for studying post-translational modification of this enzyme.
3. Semi-quantitative analysis after immunoelectron microscopy (i.e., immunogold labeling) revealed that the alveolar macrophages and type 2 pneumocytes in rat lungs were rich in cellular GPX.The intracellular localization of cellular GPX in those tissues was similar to that observed previously in rat hepatocytes.
4. By transient transfection of cellular GPX to … More Cos-7 cells, the expression was enhanced 3 times, with translocation of the enzyme to both mitochondria and nucleus.
5. By permanent expression of GPX in a human tumor cell line, the enzyme level was enhanced 30 times, with relative richness in mitochondria, nucleus and cytoplasmic matirx.
6. In selenium deficient rat kidneys, both cellular and plasma GPX were decreased in a similar fashion. The expression was inhibited at multiple steps : i.e., pre-translational, translational and posttranslational levels. The study also revealed that plasma GPX was expressed in the proximal tubules of rat kidneys and was almost completely secreted to the plasma.
7. By collaboraion with various institutions, the pathognomonic structures for familial amyotrophic lateral sclerosis in anterior horn neurons of the autopsied sampled were found to be rich in immunoreactive CuZnSOD,as stained by our polyclonal antibodies. Contribution of proteolytic mechanism mediated by ubiquitin to pathogenesis was suggested. The findings supported the gain-of-function theory for the mutant CuZnSOD to cause degenerative changes. Less

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (35 results)

All Other

All Publications (35 results)

  • [Publications] Asayama K.: "Imnunohistochemical localization and quantitive analysis of cellular glutathione peroxidase in foetal and neonatal rat tissues;fluorescence microscopy image analysis." Histochem J.28. 63-71 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K.: "Immunolocalization of cellular glutathione peroxidase in adult rat lungs and quantitative analysis after post-embedding immunogold labeling." Histochem Cell Biol.105. 383-389 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K,: "Immunohistochemical study on superexide dismutoses in spinal cords from autopside patients with amyotrophic lateral sclerosis." Dev Neurosci. 18. 492-4 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K.: "Immunohistochemical localization and quantitative analysis of superoxide dismutases in fetal and neonatal rat tissues:Fluorescence microscopy image analysis." Acta Histochem Cytochem. 29(4). 289-297 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K.: "Increased plasma cholesteryl cster transfer activity in obese children" Atherosclerosis. 129. 53-58 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K.: "New age-adjusted measure of body fat distribution in children and adolescents:standardization of waist hip ratio using multivariate analysis." Int J.Obes.21. 594-599 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K,Hayashibe H,Mishiku Y,Honda M,Ito H,Nakazawa S: "Increased activity of plasma cholesteryl ester transfer protein in chidren with end-stage renal disease receiving contnuous ambulatory peritoneal dialysis." Nephron. 72. 231-236 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K,Dobashi K,Kawada Y,Nakane T,Kawaoi A,Nakazawa S: "Immunohistochemical localization and quantitative analysis of cellular glutathione peroxidase in foetal and neonatal rat tissues : fluorescence microscopy image analysis." Histochem. J28. 63-71 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Shibata N,Hirano A,Kobayashi M,Siddique T,Deng HX,Hung WY,Kato T,Asayama K: "Intense spueroxide dismutase-1 immunoreactivity in intracytoplasmic hyaline inclusions of familial amyotrophic lateral sclerosis with posterior column involvement." J.neuropathol Exp Neurol. 55 (4). 481-490 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K,Yokota S,Dobashi K,Kawada Y,Nakane T,Kawaoi A,Nakazawa S: "Immunolocalization of cellular glutathione perosidase in adult rat lungs and quantitative analysis after post-embedding immunogold labeling." Histochem Cell Biol. 105. 383-389 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Shibata N,Asayama K,Hirano A,Kobayashi M: "Immunohistochemical study on superoxide dismutases in spinal cords from autopsied patients with amyotrophic lateral sclerosis." Dev Neurosci. 18. 492-498 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kawada Y,Asayama K,Dobashi K,Nakane T,Hayashibe H,Kawaoi A,Nakazawa S: "Immunohistochemical localization and quantitative analysis of superoxide dismutases in fetal and neonatal rat tissues : Fluorecence microscopy image analysis" Acta Histochem Cytochem. 29 (4). 289-297 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hayashibe H,Asayama K,Nakane T,Uchida N,Kawada Y,Nakazawa S: "Increased plasma cholesteryl ester transfer activity in obese children" Atherosclerosis. 129. 53-58 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nishiyama K,Murayama S,Nishimura Y,Asayama K,Kanazawa I: "Superoxide dismutase-like immunoreactivity is present in spheroids of Hallervorden Spatz disease." Acta Neurophathol.93. 19-23 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K,Hayashi K,Kawada Y,Nakane T,Uchida N,Hayashibe H,Kawasaki K,Nakazawa S: "New age-adjusted measure of body fat distribution in children and adolescents : standardization of waist hip ratio using multivariate analysis." Int J Obes. 21. 594-599 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kato S,Hayashi H,Nakashima K,Nanba E,Kato M,Hirano A,Nakano I,Asayama K,Ohama E: "Pathological characterization of astrocytic hyaline inclusions in familial amyotrophic lateral sclerosis." Am J Pathol. 151. 611-620 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K,nakane T,Uchida N,Hayashibe H,Nakazawa S,Kato K,Qin P,Gross AJ: "Age-adjusted anthropometric index of overweight optimized for ages ranging from 6 to 14 years." Clinical Pediatric Endocrinology. 6 (1). 39-45 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Noak H,Lindenau J,Rothe F,Asayama K,Wolf G: "Differential expression of superoxide dismutase isoforms in neuronal and glial compartments in the course of excitotoxically mediated" Glia. (in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Singh AK,Dobashi K,Gupta MP,Asayama K,Singh I,Orak JK: "Manganese superoxide dismutase in rat liver peroxisomes : Biochemical and immunochemical evidence." Biochim Biophys Acta. (in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nakane T,Asayama K,Kodera K,Hayashibe H,Uchida N,Nakazawa S: "Effect of selenium deficiency on cellular and extracellular glutathione peroxidases : immunochemical detection and mRNA analysis in rat kidney and serum." Free Radic Biol Med. (in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K,Sandhir R,Sheikh FF,Hayashibe H,Nakane T,Singh I: "Activation of fatty acid oxidation and enhanced expression of peroxisome proliferator-activated receptor-a in diabeticrat liver." Mol Cell Biochem. (under consideration).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Asayama K.: "Immunohistochemical localization and quantitive analysis of cellular glutathion peroxidase in foetal and neonatal rat tissues;fluorescence microscopy image analysis." Histochem J.28. 63-71 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Asayama K.: "Immunolocalization of cellular glutathione peroxidase in adult rat lungs and quantitative analysis after post-embedding immunogold labeling." Histochem Cell Biol. 105. 383-389 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Asayama K.: "Immunohistochemical study on superoxide dismutases in spinal cords from autopside patients with amyotrophic lateral sclerosis." Dev.Neurosci. 18. 492-498 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Asayama K.: "Immunohistochemical localization and quantitative analysis of superoxide dismutases in fetal and neonatal rat tissues:Fluorescence microscopy image analysis" Acta Histochem Cytochem. 29(4). 289-297 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Asayama K.: "Increased plasma cholesteryl ester transfer activity in obese chidren." Atherosclerosis. 129. 53-58 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Asayama K.: "New age-adjusted measure of body fat distribution in children and adolescents:Standardization of waist hip ratio using multivariate analysis." Int J.Obes.21. 594-599 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 朝山,光太郎: "Increased activity of plasma cholesteryl ester transfer protein in children with end-stage renal disease receiving continuous ambulatory peritonealdialysis." Nephron. 72. 231-236 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 朝山,光太郎: "Immunohistochemical localization and quantitative analysis of cellular glutathione peroxidase in foetal and neonatal rat tissues : fluorescence microscopy image analysis." Histochemical Journal. 28. 63-71 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 柴田,亮行: "Intense superoxide dismutase-1 immunoreactivity in intracytoplasmic hyaline inclusions of familial amyotrophic lateral sclerosis with posterior column involvement." Journal of Neuropathology and Experimental Neurology. 55(4). 481-490 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 朝山,光太郎: "Immunolocalization of cellular glutathione peroxidase in adult rat lungs and quantitative analysis after post-embedding immunogold labeling." Histochemistry and Cell Biology. 105. 383-389 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 柴田 亮行: "Immunohistochemical study on superoxide dismutases in spinal cords from autopsied patients with amyotrophic lateral sclerosis." Developmental Neuroscience. 18. 492-498 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 朝山 光太郎: "活性酸素と病態,糖尿病" 臨床検査. 40(2). 176-180 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 川田 康介: "Immunohistochemical localization and quantitative analysis of superoxide dismutases in fetal and neonatal rat tissues : Fluorescence microscopyimage analysis." Acta Histochemica et Cytochemica. 29(4). 289-297 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 西山 和利: "Superoxide dismutase-like immunoreactivity is present in spheroids of Hallervorden Spatz disease." Acta Neuropathologica. (印刷中). (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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