Analysis of Synaptic Connections at the Electron Microscopic Level Using Viral Vectors

  • Takahiro Furuta
  • Keiko Okamoto-Furuta
  • Hiroyuki Hioki
Part of the Neuromethods book series (NM, volume 110)


Anterograde tracing using Sindbis virus vector resolves several problems of conventional tracers. Usage of the Sindbis virus as a neural tracer prevents the axons passing through the injection site from being labeled because the virus does not infect from axons. Combining the methodological merit of the virus tracer with electron microscopic technique, we analyze morphological characteristics of ultrastructure of axon terminals which are provided specifically from the region of interest, and thus we can systematically extract information of synaptic organization from complicated neural networks. Here, we introduce a method where Sindbis virus labeling is combined with an immunoelectron microscopic technique. Using this technique, we observe synaptic contacts and GABA immunoreactivity in ultrastructure of axon terminals which are revealed to be projected from the virus injection site.


Sindbis virus Anterograde tracing Electron microscopy GFP Post-embedding immunostaining 



This work was supported by the Ministry of Education, Science, Sports, and Culture of Japan Grants (15H04266, 15H01430 and 15K14333). We thank Dr. H. Bokor for the helpful advice on post-embedding immunostaining methods.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Takahiro Furuta
    • 1
  • Keiko Okamoto-Furuta
    • 2
  • Hiroyuki Hioki
    • 1
  1. 1.Department of Morphological Brain ScienceGraduate School of Medicine, Kyoto UniversityKyotoJapan
  2. 2.Division of Electron Microscopic StudyCenter for Anatomical Studies, Graduate School of Medicine, Kyoto UniversityKyotoJapan

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