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1993 Fiscal Year Final Research Report Summary

Mechanism of new regeneration activating factor of aged nervous tissue secreted from hepatocytes and its determination with vitro immunization

Research Project

Project/Area Number 04836020
Research Category

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

Allocation TypeSingle-year Grants
Research Field 老化(加齢)
Research InstitutionYokohama City University

Principal Investigator

HORIE Hidenori  横浜市立大学, 医学部, 講師 (80046135)

Project Period (FY) 1992 – 1993
Keywordstissue culture / collagen gel / neural regeneration / adult murine / hepatocyte / enhancement factor / retina / dorsal root ganglion
Research Abstract

We have established culture system of central and peripheral nervous tissues of matured or aged murine by introduction of three-dimensional collagen gel culture. Retina was used as a central nervous tissue and dorsal root ganglion was as a peripheral one. Hepatocyte secreted factor was applied to these cultures to demonstrate its activity, mechanism, and determination of the factor enhancing neural regeneration. Hepatocytes were dissected from adult rats by collagenase perfusion method and cultured in a serum-free William's E medium. The culture medium was applied to cultured retina explants as hepatocyte conditioned medium(HCM). Retina was removed from an adult rat 7 days after optic nerve transection and dissected to small fragment. They were embedded in collagen gel and cultured in the same medium as that in hepatocytes. Processes appeared 24 h after in culture. Immunocytochemical staining revealed that these processes were positive either to anti-neurofilament antibody or anti-thy-1.2 antibody. These results clarified that regenerating processes were neurites from retinal ganglion cells. When HCM was applied to a retina explant, the number of regenerating neurites increased to 2.6 times larger than that in control medium. This enhancement activity appeared in 5k< <100k fraction after membrane filtration and disappeared after the treatment with 95゚C, 5 min. This factor enhanced neurite regeneration from transected nerve terminals of adult or aged murine dorsal root ganglion (DRG), too. Since the explants of retina or DRG kept structures and functions similar to the tissues in situ, this factor is expected to work in vivo as a neural regeneration activating factor. We have already examined 20 substances recognized as a growth factor or a neural regeneration activating one, but no one exerted the same effect as a hepatocyte secreted one. This factor is thought to be new. We have developed purification of the substance and it will reach a final stage to decide the

  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Sango,K.,Horie,H.,Takano,M.,Inoue,S.,Takenaka,T.: "Diabetes-induced reduction of neuronal survival in hypotonic environments in culture" Brain Research Bulletin. (in press). (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Horie,H.,Ito,S.,Takano,M.: "Regional difference in neurite regeneration of postnatal retinal explants." Brain Research Bulletin. (in press). (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hikawa,N.,Horie,H.,Takenaka,T.: "Macrophage-enhanced neurite regeneration of adult dorsal root ganglia neurons in culture" NeuroReport. 5. 41-44 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Horie,H.,Sango,K.,Takano,M.,Takenaka,T.: "Response of aged neurons to super-hypotonic environment." Japanese J.Physiology. 43 Suppl.1. S193-196 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sango,K.,Horie,H.Inoue,S.,Takamura,K.,Takenaka,T.: "Age-related changes of DRG neuronal attachment to extracellular matrix proteins in vitro." NeuroReport. 4. 663-666 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Chi,H.,Horie,H.,Hikawa,N.,Takenaka,T.: "Isolation and age-related characterization of mouse Schwann cells from dorsal root ganglion explants in type I collagen gels." J.Neurosci.Res.35. 183-187 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 堀江秀典: "神経細胞膜の物理的特性の加齢変化,14巻 発生・分化および老化" 東京化学同人, (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sango, K., Horie, H., Takano, M., Inoue, S.and Takenaka, T.: "Diabetes-induced reduction of neuronal survival in hypotonic environments in culture." Brain Res.Bull.in press. (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Horie, H., Ito, S.and Takenaka T.: "Regional differences in neurite regeneration of postnatal retinal explants." Brain Res.Bull.in press. (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hikawa, N., Horie, H.and Takenaka T.: "Macrophage-enhanced neurite regeneration of adult dorsal root ganglia neurons in culture." Neuro Report. 5. 41-44 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Horie, H., Akahori, Y.and Kadoya, T.: "Neurotrophic factors for peripheral neural regeneration : Hepatocyte secreted factor as a new one." Tiss.Cul.Commun.12. 229-236 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Horie, H., Sango, K., Takano, M.and Takenaka, T.: "Response of aged neurons to super-hypertonic environment." Japanese J.Physiol.43, Suppl.1. 193-196 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sango, K., Horie, H., Inoue, S., Takamura, K.and Takenaka, T.: "Age-related changes of DRG neuronal attachment to extracellular matrix proteins in vitro." Neuro Report. 4. 663-666 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Chi, H., Horie, H., Hikawa, N.and Takenaka, T.: "Isolation and age-related characterization of mouse Schwann cells from dorsal root ganglion explants in type I collagen gels." J.Neurosci. Res.35. 183-187 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Horie, H.: "Establishment of neuronal tissue culture method and its application." Tiss.Cult.Res.Commun.11. 361-368 (1992)

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

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Published: 1995-03-27  

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