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Mobility of neuronal growth-cones and formation of synapses

Research Project

Project/Area Number 08044257
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research Field Neuroscience in general
Research InstitutionTokyo Medical and Dental University

Principal Investigator

KATAYAMA Yoshifumi  Tokyo Med & Dent Univ, Med Res Inst, Prof, 難治疾患研究所, 教授 (20014144)

Co-Investigator(Kenkyū-buntansha) TATSUMI Hitoshi  Nagoya Univ, Sch Med Assit Prof, 医学部, 助手 (20171720)
FURUKAWA Tetsushi  Tokyo Med & Dent Univ, Med Res Inst, Assist Prof, 難治疾患研究所, 助手 (80251552)
HIRAI Kenji  Tokyo Med & Dent Univ, Med Res Inst, Assoc Prof, 難治疾患研究所, 助教授 (70156628)
MCCAING C.D.  Univ Aberdeen, Biomed Sci, Senior Lecturer, 医学部, 上級講師
TSUJI S.  P. &M. キュリー大学, CNRS, 教授
TSUJI Shigeru  P & M Curie Univ, CNRS,Research Director
MCCAIG C. D.  アバディーン大学, 医学部, 上級講師
MCCAIG C.D.  アバディーン大学, 医学部, 準教授
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥8,600,000 (Direct Cost: ¥8,600,000)
Fiscal Year 1997: ¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 1996: ¥4,300,000 (Direct Cost: ¥4,300,000)
KeywordsNeuron / Growth cone / Synapse / Neurotransmitter / Optical technology / Ion channel / Electric field / Mechanosensitivity / 光学技術 / 光ピンセット / 近接場光学顕微鏡 / カルシウムチャネル / 神経成長円錐 / 神経伝達物質 / カルシウム / 神経成長因子 / ホールセルパッチクランプ
Research Abstract

It is necessary to apply recent advance in optical technology to studying cellular physiology. Using some new techniques, we demonstrated that growth cones of rat cultured neurons can release neurotransmitter and be sensitive to both mechanical and electric field stimulation.
Glutamate release from growth cones was detected by using an acutely dissociated hippocampal neuron as a biodetector from which whole cell patch clamp recordings were made. The release was dependent on Ca^<2+> and was abolished by omega-agatoxin. Cultured neurons were pretreated with the toxin, then incubated with the antibody to the toxin and further stained with gold-conjugated antibody. Since clusters of immuno-gold particles were observed over the membrane of growth cones by means of scanning and transmission electron microscpes, it is suggested that omega-agatoxin-sensitive Ca^<2+> channels were immunologically localized on growth cones.
The mechanism underlying cathodally-directed growth of cultured DRG neurit … More es was investigated by using confocal laser scanning microscopy to observe cytoskeletal elements, second messenger signaling elements and the nucleus in neurons exposed to small physiological electric fields. Three types of fluorescent probes were used, a) Rhodamine halloidin stains filamentous actin, b) a fluorescein-conjugated monoclonal antibody labelled proteins phosphorylated on tyrosine residues and c) a marker for mitochondria clearly delineated the position of the nucleus by exclusion, being present only in the cytoplasma. Physiological electric fields induce asymmetries of the filamentous actin cytoskeleton and proteins phosphorylated on tyrosine residues in growth cones, and alter the positioning of cell nucleus.
When mechanical stimulation was applied to growth cones of DRG neurons, inward currents were recorded, from their soma at the holding potential of -60mV,using whole cell patch clamp technique. Almost no electrical response was recorded when other sites except lamellipodium were stimulated. The outward currents were supposed to be due mainly to activation of Cl^- channels according to their electrophysiological properties.
Evanescent light illumination was introduced into a high resolution video-enhanced differential interference contrast microscope. Growth cones and soma of neurons which were cultured on glass plates were stained with a fluorescent dye, Dil, and were observed with an internal total reflection evanescent light microscope. The evanescent images showed that growth cones and some limited areas of soma attached to the surface of culture glass plate. 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] Hirai K,Katayama Y,Misu Y: "L-DOPA induces concentration-dependent facilitation and inhibition of presynaptic ACh release in the guinea-pig.." Brain Research. 718. 105-111 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Furukawa T et al.(other 4 authors): "Functional linkage of the cardiac ATP-sensitive K^+ channel to the actin cytosleleton." Pflugers Archiv. 432. 504-512 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Rajnicek AM,Britland S,McCaig CD: "Contact guidance of CNS neurites on grooved quart:influence of groove dimensions,neuronal age and cell type." Journal of Cell Science. 110. 2905-2913 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Soeda H,Tatsumi H,Katayama Y: "Neurotransmitter release from growth cones of the dorsal root ganglion neurons in culture." Neuroscience. 77. 1187-1197 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Furukawa T,Ogura T,Katayama Y,Hiraoka M: "Characteristics of rabbit ClC-2 current expressed in Xenopus oocytes and its contribution to volume regulation." Americal Journal of Physiology. (in press). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Katayama Y,Morita K, Hirai K: "Concentration-dependent actions of a new indene derivative,TN-871,in the enteric nervous system." European Journal of Pharmacology. (in press). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] McCaig C D(Ed): "Nerve growth and guidance" Portland Press,London, 170 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hirai K,katayama Y,Misu Y.: "L-DOPA induce concentration-dependent facilitation and inhibition of presynaptic acetylcholine release in the guineapig submucous plexus." Brain Research. 718. 105-111 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Morita K,Katayama Y.: "Tetraethylammonium-sensitive calcium-senstitive potassium current in a sub-class of the bullfrog dorsal root ganglion cells." Neurosci.Lett.215. 193-196 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Furukawa T,Yamane T,Terai T,Katayama Y,Hiraoka M.: "Functional linkage of the cardiac ATP-sensitive K^+ channel to the actin cytoskeleton." Pflugers Archiv. 432. 504-512 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Uma Maheswari R,Mononobe S,Tatsumi H,Katayama Y,Ohtsu M.: "Observation of subcellular structures of neurons by an illumination mode near-field optical microscope under an optical feedback control." Optical Review. 3. 463-467 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Stewart R,McCaig CD.: "Lectins implicate specific carbohydrate domains in electric field stimulated nerve growth and guidance." J.Neurobiology. 30. 425-437 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Britland S,McCaig CD.: "Embryonic Xenopus neurites integrate and respond to simultaneous electrical and adhesive guidance cues." Exp.Cell Research. 226. 31-38 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Erskine L,McCaig CD.: "Integrated interactions between chondroitin sulphate proteoglycans and weak dc electric fields regulate nerve growth cone guidance in vitro." J.Cell Science. 110. 1957-1965 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Rajnicek AM,Britland S,McCaig CD.: "Contact guidance of CNS neurites on grooved quartz : influence of groove dimensions, neuronal age and cell type." J.Cell Science. 110. 2905-2913 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Rajnicek AM,McCaig CD.: "Guidance of CNS growth cones by substratum grooves and rides : effects of inhibitors of the cytoskeleton, calcium channels and signal transduction pathways." J.Cell Science. 110. 2915-2924 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] McCaig CD,Zhao M.: "Physiological electrical fields modify cell behavior." BioEssays. 19,09. 819-826 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Soeda H,Tatsumi H,Katayama Y.: "Neurotransmitter release from growth cones of the dorsal root ganglion neurons in culture." Neuroscience. 77. 1187-1199 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Anglade P,Tsuji S.: "Hundredth anniversary of the "synapse" : I.A short history of the milestones in synapse research." Zoological Science. 14. 533-537 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tsuji S,Anglade P.: "Hundredth anniversary of the "synapse" : II.Study of the cholinergic synapse." Zoological Science. 14. 539-548 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hirai K,Browning KN,Lees GM.: "Neuropeptide Y hyperpolarizes submucosal neurons of the guinea-pig descending colon." Neurosci.Letters. 227. 212-214 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Furukawa T,Ogura T,Katayama Y,Hiraoka M.: "Characteristics of rabbit CIC-2 current expressed in Xenopus oocytes and its contribution to volume regulation." American J.Physiology. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Katayama Y,Morita K,Hirai K.: "Concentration-dependent actions of a new indene derivative, Tn-871, in the enteric nervous system." European J.Pharmacology. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Soeda H, Tatsumi H, Katayama Y: "Neurotransmitter release from growth cones of the dorsal root ganglion neurons in culture." Neuroscience. 77. 1187-1199 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hirai K, Browning KN, Lees GM: "Neuropeptide Y・hyperpolarizes submucosal neurons of the guinea-pig descending colon." Neuroscience Letters. 277. 212-214 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Rajnicek AM, Macaig CD: "Giudance of CNS neurites by substratum grooves and didges; II. Effects of inhibitors of ...." Journal of Cell Science. 110. 2915-2924 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Tsuji S, Anglade P: "Hundredth anniversary of the synapse; I. Ashort history of the milestones in synapse research." Zoological Science. 14. 533-537 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] FurukawaT, Ogura T, Katayama Y, HiraokaM: "Characteristica of rabbit C1C-2 expressed in Xenopus oocytes and its contribution to volume regulation." Americal Journal of Physiology. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Katayama Y, Hirai K, Morita K: "Concentration-dependent actions of a new indene derivative TN-871 in the enteric nervous system." European Journal of Pharmacology. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hirai,K.,Katayama,Y.and Misu,Y.: "L-DOPA induces concentration-dependent facilitation and inhibition of presynaptic acetylcholine relrase in the guinea-pig submucous plexus." Brain Res.718. 105-111 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Mosita,K.and Katayama,Y.: "Tetraethylammonium-sensitive calcium-sensitive pottasium current in a subclass of the bullfrog dorsal root ganglion cells." Neuroscience Letters. 215. 193-196 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Furukawa,T.,Yamane,T.,Terai,T.,Katayama,Y.and Hiraoka,M.: "Functional linkage of the cardiac ATP-sensitive K^+ channel to the actin cytoskeleton." Pflugers Arch.431. 504-512 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Stewart R.,Allan,D.W.and McCaig,C.D.: "Lectins implicate specific carbohydrate domains in electric field stimulated nerve growth and guidance" J.Neurobiol.30. 425-437 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Britland,S.and McCaig,C.D.: "The response of cultured Xenopus neurites to simultaneous electrical and adhesive guidance cues." Exp.Cell Res.226. 31-38 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Soeda,H.,Tatsumi,H.and Katayama,Y.: "Neurotransmitter release from growth cones of rat dorsal root ganglion neurons in culture." Neuroscience. (in Press). (1997)

    • Related Report
      1996 Annual Research Report

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

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