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1990 Fiscal Year Final Research Report Summary

Molecular Biological Studies on the Etiology of Neurodegenerative Diseases.

Research Project

Project/Area Number 01480240
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Neurology
Research InstitutionNiigata University

Principal Investigator

TSUJI Shoji  Hospital of Medical School, Department of Neurology, Associate, 医学部附属病院, 助手 (70150612)

Project Period (FY) 1989 – 1990
Keywordssubtractive cloning / Neurodegenerative diseases / cDNA library / Nervous system / Brain / Spinal cord / Molecular cloning / Linkage analysis
Research Abstract

Neurodegenerative disorders are characterized by selective loss of particular types of neurons. The identification of genes which are expressed in specific neurons such as motor neurons would bring us better understanding of differentiation of neurons as well as the molecular mechanisms of selectiveloss of the neurons.
I have developed a method for isolation of gene which are expressed in particular types of neurons employing subtractive cloning. For the strategy it is crucially important to isolate undegraded RNAs from post-motem brains and spinal cords. First, the stability of messenger RNA in Post-mortem human bains and spinal cords was investigated. It was concluded that messenger RNAs are not degraded as long as 12 hours. Complementary DNA libraries were made from postmortem human brain and spinal cord Analysis of the cDNA libraries have shown the presence of full-length cDNA clones for neuron-specific olase, S100 and myelin-associated glycoprotein.
PolyA (+) RNA from human spinal cord was labeled with ^<32>P and reassociated in phenol-emulsion with human brain cDNAs to remove the commonly expressed genes. The human spinal cord cDNA library was screened with the subtracted ^<32>P-labeled probe, and I have succeeded in isolation of four cDNA clones. Northern blot analysis revealed that increased expression of the two cDNA clones. The results suggest a potential feasibility of the strategy for identifying genes expressed in selective neurons.
To identify genes involved in the development of juvenile-Parkinsonism, a neuro-degenerative disorder, linkage analysis on tyrosine hydroxylase locus was performed, as previous studies suggested tyrosine hydroxylase is a candidate gene for juvenile Parkinsonism. Linkage analysis employing tyosine hydroxylase gene and adjacent VNTR (variable number of tandem repeat) probes including H-RASI and INS has clearly demonstrated that tyrosine hydroxylase gene is not linked to juvenile Parkinsonism.

  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Hisashi Kobayashi: "Stability of messenger RNA in postmortem human brains and constrction of human brain cDNA libraries." J.Mol.Neucosci.2. 29-34 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hajime Tanaka: "Linkage analysis of juvenile parkinsonism to tyrosine hydroxylase gene locus on chromosome ll." Neurol.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ryoko Koike: "Physiological significance of fatty acid elongation system in adrenoleukodystrophy." J.Neurol.Sei.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Makoto Yoneda: "A common mitochondrial DNA mutation in the tーRNA^<LU5> of patients with myoclonus epilepsy associated with raggedーred fibers." Biochem.Int.21. 789-796 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toyoaki Yamauchi: "Molecular cloning of two species of cDNAs for Human αーNーacetylgatactosaminidase and expression in mammalian cells." Biochem.Biophys.Res.Commun.170. 231-237 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shoji Tsuji: "Molecular cloning of a follーlength cRNA for human αーNーacetylgalactosaminidase (αーgalactosidase B)." Biochem.Biophys.Res.Commun.163. 1498-1504 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Makoto Yoneda: "Mitochondrial mutation in family with Leber's hereditary optic neuropathy." Lancet. 1. 1076-1077 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Rui Kondo: "Identification of a mutation in arylsulfatase A gene of a patient with adult type metachromatic leukodystrophy." Amer J. Hum. Genet.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hisashi Kobayashi,: "Stability of messenger RNA in postmortem human brains and construction of human brain cDNA libraries." J. Mol. Neurosci.,. 2,. 29-34 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hajime Tanaka,: "linkage analysis of juvenile Parkinsonism to tyrosine hydroxylase gene locus on choromosome 11." Neurol.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ryoko Koike,: "Physiological significance of fatty acid elongation system in adrenoleukodystrophy" J. Neurol. Sci.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Rui Kondo,: "Identification of a mutation in aryl sulfatase gene of a patient with adult type metachromatic leukodystrophy" Amer. J. Hum. Genet.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Makoto Yoneda,: "A common mitochondrial DNA mutation in the t-RNA^<Lys> of patients with myoclonus epilepsy associated with ragged-red fibers." Biochem. Int.21,. 789-796, (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toyoaki Yamauchi,: "Molecular cloning of two species of -N-acetylgalactosaminidase and expression in mammalian cells." Biochem. Biophys. Res. Commun.170. 231-237 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shoji Tsuji,: "Molecular cloning of a full-length cDNA for human -N-acetylgalactosaminidase (-galactosidase B)." Biochem. Biophys. Res. Commun.163,. 1498-1504 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Makoto Yoneda,: "Mitochondrial mutation in family with Leber's hereditary optic neuropathy." Lancet. 1,. 1076-1077, (1989)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-08-12  

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