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1995 Fiscal Year Final Research Report Summary

Micro-quantification of cell-cycle regulating factors by Enzyme-Linked Immuno-Cycling Assay

Research Project

Project/Area Number 06808083
Research Category

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

Allocation TypeSingle-year Grants
Research Field Neurochemistry/Neuropharmacology
Research InstitutionYokohama City University School of Medicine

Principal Investigator

NAKAYAMA Takashi  Yokohama City University, Lecturer, 医学部, 講師 (90150060)

Co-Investigator(Kenkyū-buntansha) KATO Takahiko  Yokohama City University, Professor, 医学部, 教授 (80010023)
Project Period (FY) 1994 – 1995
Keywordscyclin / cdk / retina / nitric oxide / cerebellum
Research Abstract

Neural cells must develop to mature after free from cell-cycle and never divide again. Despite these facts, cell-cycle regulating factors, i.e.cyclins and cyclin-dependent kinases (cdk) are shown to be exist in post-mitotic neurons. To clarify the physiological roles of those molecules in neuron, we first intended to identify the molecular species of cyclin and cdk in embryonic chick retina. The immunohistochemical studies using cultured retinal neurons revealed that antibodies raised against cyclin D1 or cdk5 were immunoreactive to all the kind of retinal neurons including immature photoreceptor cells. And then next we examined developmental change of cyclin D1 and cdk5 expression in rat cerebellum by immunoblotting. It was found that cyclin D1 was consistently expressed after birth to adulthood, however expression of cdk5 was transient around 8-days-old where the layr formation of cerebellum is extremely remarkable. These results suggest that cdk5 may be involved in the maturation of granule cells in cerebellum. We also found that nitric oxide (NO) is necessary for the maturation and migration of granule cells during the layr formation of the cerebellum. When nitric oxide synthase (NOS) was irreversibly inhibited, migration of granule cells was drastically decreased, and consequently perturbation of the layr formation occurred. To identify the subclass of NOS responsible for this phenomenon, about 1 to 3 mug in dry weight of tissue from external granular layrs, molecular layrs and internal granular layrs was electrophoresed and analyzed by micro-immunoblotting. Micro-analysis revealed that neuronal-type NOS and also endothelial-type NOS were expressed in each layrs during the layr formation.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Wang, W. Inoue, N. Nakayama, T.: "An assay method for nitric oxide synthase in crude samples by determining product NADP^+" Anal. Biochem.227. 274-280 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Wang, W. Inoue, N. Nakayama, T.: "Nitric oxide synthase (NOS) supports developmental and differentiation of neurons in cerebellar layers from rats." J. Neurochem.65. S215A (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Furuya, S., Nagata, R., Ozaki, Y., Furuya, K., Nakayama, Y. Nagahama,: "A monoclonal antibody to astrocytes, subepithelial fibroblasts of small intestinal villi and interstitial cells of myenteric plexus." Anat. Embryol.(in press). (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamaguchi, K., Nakayama, T. Akagawa, K.: "Enhancement of neurite-sprouting by suppression of HPC-1/Syntaxin-1A activity in the cultured vertebrate nerve cells." Brain Res.(in press). (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 五嶋 良郎、中山 孝: "反発因子の細胞内情報伝達" 細胞工学. 15(2). 181-189 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Wang, W.Inoue, N.Nakayama, T.: "An assay method for nitric oxide synthase in crude samples by determining product NADP+." Anal.Biochem.227. 274-280 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wang, W.Inoue, N.Nakayama, T.: "Nitric oxide synthase (NOS) supports developmental and differentiation of neurons in cerebellar layrs from rats." J.Neurochem.65. S215A (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Inoue, N.Soga, T.Nakayama T.and Kato T.: "Expression and regulation of sodium pump isoforms in cultured cerebellar granule cells." J.Neurochem.65. S159D (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kato, T.and Kijima, M.: "Microanalysis of S-antigen distribution in ocular tissues and retinas from rats." Neurochem.Res.20. 331 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wang, W.et al.: "Nitric oxide synthase (NOS) activities in developing and differentiating cerebellar layrs from rats." Neurochem.Res.20. 365 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Furuya, S., Nagata, R., Ozaki, Y., Furuya, K., Nakayama, T.and Nagahama, M.: "A monoclonal antibody to astrocytes, subepithelial fibroblasts of small intestinal villi and interstitial cells of myenteric plexus." Anat.Embryol.(in press). (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamaguchi, K., Nakayama, T.and Akagawa, K.: "Enhancement of neurite-sprouting by suppression of HPC-1/Syntaxin-1A activity in the cultured vertebrate nerve cells." Brain Res.(in press). (1996)

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

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Published: 1997-03-04  

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