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Visualization of Thalamocortical Axon Branching and Synapse Formation in Organotypic Cocultures
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Neurociência
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JoVE Journal Neurociência
Visualization of Thalamocortical Axon Branching and Synapse Formation in Organotypic Cocultures

Visualization of Thalamocortical Axon Branching and Synapse Formation in Organotypic Cocultures

DOI:
10.3791/56553-v

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06:16 min

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March 28, 2018

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,
1Department of Medical Neuroscience, Graduate School of Medical Sciences,Kanazawa University, 2Neuroscience Laboratories, Graduate School of Frontier Biosciences,Osaka University

Capítulos

  • 00:04Título
  • 00:58Preparation of Cortical Slices
  • 02:24Procedure of Electroporation
  • 04:37Results: Thalamocortical Axons Coculture and Time-lapse Imaging
  • 05:38Conclusion

Summary

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This protocol describes a method for simultaneous imaging of thalamocortical axon branching and synapse formation in organotypic cocultures of the thalamus and cerebral cortex. Individual thalamocortical axons and their presynaptic terminals are visualized by a single cell electroporation technique with DsRed and GFP-tagged synaptophysin.

Tags

ThalamocorticalAxon BranchingSynapse FormationOrganotypic CoculturesNeural Circuit FormationAxon GrowthSynapse FormationCerebral CortexPostnatal Day-two RatPrimary Visual CortexSomatosensory CortexCulture InsertThalamic BlockE15 EmbryoGlass CapillariesElectrode PipettePlasmid Injection

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