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Identification of dopaminergic neuron trophic factor using subtraction hybridization and bioassay

Research Project

Project/Area Number 03454133
Research Category

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

Allocation TypeSingle-year Grants
Research Field Neurophysiology and muscle physiology
Research InstitutionNagoya City University

Principal Investigator

NISHINO Hitoo  Nagoya City Univ., Physiology, Professor, 医学部, 教授 (60073730)

Co-Investigator(Kenkyū-buntansha) HASHITANI Takeshi  Nagoya City Univ., Physiology, Instructor, 医学部, 助手 (30172852)
FURUYAMA Fujiya  Nagoya City Univ., Physiology, Instructor, 医学部, 助手 (00080101)
Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1992: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1991: ¥5,600,000 (Direct Cost: ¥5,600,000)
KeywordsDopamine / Trophic factor / Neural transplantation / Parkinson's disease / Gene expression / ドーパミン細胞 / ド-パミンニュ-ロン / 栄素因子 / ハイブリダイゼ-ション / バイオアッセイ / mRNA / cDNA / 線条体 / パ-キンソン病
Research Abstract

Using bioassay and subtraction hybridization, we tried to identify the trophic factor for the dopaminergic (DAergic) neurons in the DA-denervated striatum in the rat. Results are following.
1. Neural transplantation : Grafted DAergic neurons grew better in the DA-depleted striatum than in the normal striatum. Tyrosine hydroxylase immunoreactivity was higher (x 1.3) and the neurite extension was stronger (x 1.3-1.8) in the DA-depleted striatum.
2. Bioassay : Among many fractions, a fraction with molecular weight of 13 to 45 kD in the tissue extract from the DA-depleted striatum enhanced most strongly the survival and neurite extension of the cultured DAergic neurons. This effect was stable with or without astrocyte feeder layers. It also stimulated the neurite genesis in PC12D with the enhancement of the slow activating/long-lasting Ca current. By the treatment of antibodies for NGF and FGF the effect was reduced down to about 50%.
3. Antibody : So far antibodies with strong titers have not been obtained yet.
4. Gene expression : Using lambdagt 10 as a vector, we got cDNA library from the DA-depleted striatal tissue. After the differential hybridization of these cDNAs with the mRNAs from the intact and DA-depleted striatum we identified 54 plaques whose mRNA expression was much higher in the DA-depleted striatum. The enhancement of mRNA expression was reconfirmed in at least 6 plaques by Sourthern blotting and in situ hybridization. After sequencing these DNAs, three were identified as cytochrome oxidase subunit-I, alpha-globin and ribosomal protein S-8. Other three were found to be unknown substances.
Thus, we got several candidates for trophic factor for DAergic neurons. The purification and characterization of these substances are now under the investigation.

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (29 results)

All Other

All Publications (29 results)

  • [Publications] Nishino,H.,Hashitani,T.and Kumazaki,M.: "Grafting of catecholaminergic cells in the mammalian brain and reconstruction of disturbed function" Comp.Biochem.Physiol.98C. 211-220 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nishino,H.,Hashitani,T.and Kumazaki,M.: "Effects of trophic factors on functional recovery after grafting of dopaminergic neurons in 6-OHDA lesioned rats." Adv.Neurotrauma Res.3. 1-7 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hashitani,T.,Furuyama,F.,Nishino,H.,et al.: "Sialyl cholesterol enhances the development of grafted neurons and motor recovery." Brain Res.Bull.29. 795-806 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nishino,H.,Koide,K.,Aihara,N.,et al.: "Striatal grafts in the ischemic striatum improve pallidal GABA release and passive avoidance." Brain Res.Bull.(1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nishino,H.,Aihara,N.,Czurko,A.,et al.: "Reconstruction of GABAergic transmission and behavior by striatal cell gratfs in rats with ischemic infarcts in the middle cerebral artery." J.Neural Transp.Plast.,. (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nishino,H.: "Intracerebral grafting of catecholamine producing cells and reconstruction of disturbed brain function." Neurosci.Res.,. (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Oomura,Y.,Nishino,H.,Karadi,Z.,et al.: "Advances in Physiological Sciences." Ascian and Oceanian Physiological Societies, 648-660 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nishino, H., Hashitani, T. and Kumazaki, M.: "Grafting of catecholamiergic cells in the mammalian brain and reconstruction of disturbed function : basic problems to be solved." Comp. Biochem. Phsiol.98C. 211-220 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ishida, Y., Hashiguchi, H., Yamamoto, R., Hashitani, T., Ikeda, T. and Nishino, H.: "Effect of intra-amygdala dopaminergic grafts on methamphetamine-induced locomotor activity, extracellular dopamine and dopamine metabolite overflow." Brain Research. 549. 342-345 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nishino, H., Hashitani, T. and Kumazaki, M.: "Effects of trophic factors on functional recovery after grafting of dopaminergic neurons in 6-OHDA lesioned rats." Adv. Neurotrauma Res.3. 1-7 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hashitani, T., Furuyanma, F., Kumazaki, M., Xhou, X.R., Geller, H.M., Toyomaki, Y., Okamoto, K., Oomura, Y. and Nishino, H.: "Sialyl cholesterol enhances the development of grafted neurons and motor recovery." Brain Res. Bull.29. 795-806 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nishino, H., Koide, K., Aihara, N., Kumazaki, M., Sakurai, T. and Nagai, H.: "Striatal grafts in the ischemic striatum improve pallidal GABA release and passive avoidance." Brain Re. Bull.(1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nishino, H., Aihara, N., Czurko, A., Hashitani, T., Isobe, Y., Ichikawa, O. and Watari, H.: "Reconstruction of GABAergic transmission and behavior by striatal cell grafts in the middle cerebral artery." J. Neural Transplantation & Plasticity. (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nishino, H.: "Intracerebral grafting of catecholamine producing cells and reconstruction of disturbed brain function" Neuroscience Research. (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nishino,H.,Hashitani,T.and Kumazaki,M.: "Grafting of catecholaminergic cells in the mammalian brain and reconstruction of disturbed function" Comp.Biochem.Physiol.98C. 211-220 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Nishino,H.,Hashitani,T.and Kumazaki,M.: "Effects of trophic factors on functional recovery after grafting of dopaminergic neurons in 6-OHDA lesioned rats." Adv.Neurotrauma Res.3. 1-7 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Hashitani,T.,Furuyama,F.,Nishino,H.,et al.: "Sialyl cholesterol enhances the development of grafted neurons and motor recovery." Brain Res.Bull.29. 795-806 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Nishino,H.,Koide,K.,Aihara,N.,et al.: "Striatal grafts in the ischemic striatum improve pallidal GABA release and passive avoidance." Brain Res.Bull.(1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Nishino,H.,Aihara,N.,Czurko,A.,et al.: "Reconstruction of GABAergic transmission and behavior by striatal cell grafts in rats with ischemic infarcts in the middle cerebral artery." J.Neural Transp.Plast.,. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Nishino,H.: "Intracerebral grafting of catecholamine producing cells and reconstruction of disturbed brain function." Neurosci.Res.(1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Oomura,Y.,Nishino,H.,Karadi,Z.,et al.: "Advances in Physiological Sciences." Ascian and Oceanian Physiological Societies, 648-660 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Nishino,T.Hashitani & M.Kumazaki: "Grafting of Catecholaminergic cells in the mammalian brain and reconstruction of disturbed function:basic problems to be solved." Comp.Biochem.Physiol.98C. 211-220 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] H.Nishino,S.Hattori,K.Muramoto & T.Ono: "Basal ganglia neural activity during operant feeding behavior in the monkey relation to sensory integration and motor execution." Brain Res.Bull.27. 463-468 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Y.Ishida,H.Hashiguchi,R.Yamamoto,H Nishino et al.: "Effect of intraーamygdala dopaminergic grafts on methamphetamineーinduced locomotor activity,extracellular dopamine and dopamine metabolite ovorflow : a comparison with the effect of intraーaccumbons grafts." Brain Res.549. 342-345 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Y.Oomura,H.Nishino,Z.Karadi,S.Aou & T.R.Scott: "Taste and olfactory modulation of feeding related neurons in behaving monkey." Physiol.Behav.49. 943-950 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Z.Kardi,Y.Oomura,H.Nishino,T.R.Scott et al.: "Responses of lateral hypothalamic glucoseーsensitive and glucoseーinsensitive neurons to chemical stimuli in behaving rhesus monkeys." J.Neurophysiol.

    • Related Report
      1991 Annual Research Report
  • [Publications] H.Nishino,T.Hashitani & M.Kumazaki: "Effects of trophic factors on functional recovery after grafting of dopaminergic neurons in 6ーOHDA lesioned rats." Adv.Neurotrauma Res.

    • Related Report
      1991 Annual Research Report
  • [Publications] S.Hattori,Q.M.Li,N.Matsui & H.Nishino: "Treadmill running test for evaluating locomotor activity after 6ーOHDA lesions and dopaminergic cell grafts in the rat." Brain Res.Bull.

    • Related Report
      1991 Annual Research Report
  • [Publications] T.Hashitani,F.Furuyama,H.Nishino et al.: "αーsialyl cholesterol and sialidaseーtolerant derivative,αー(3βーhydroxysialyl)cholesterol,stimulate the development of midbrain dopaminergic neurons and motor recovery following intracerebral grafting in 6ーOHDA treated rats." Brain Res.Bull.

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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