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Combined Immunocytochemistry and Tracing of Neural Connections

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Immunocytochemistry and Related Techniques

Part of the book series: Neuromethods ((NM,volume 101))

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Abstract

Neuronal tracing (neurotracing ) using anterograde and retrograde tracers is widely used to study the projections between different brain regions and the wiring between individual neurons . Neurotracing is a technique essential not only for examining the connectivity of complex neuronal networks but also for providing the neuroanatomical basis for electrophysiological , pharmacological and behavioral experiments. If neurotracing is combined with immunocytochemical labeling, the combined technique can characterize the neurochemical properties, postsynaptic targets and innervation modes of neurons. The utility and versatility of this approach can be further extended by adopting appropriate cellular and subcellular markers for immunocytochemistry , by applying the approach to animal models generated by advanced gene-manipulation technology, and by using single-cell labeling techniques, e.g., after viral transfection of fluorescent proteins or in utero /in vivo electroporation . In this chapter, we introduce the methods for combined immunocytochemistry and neurotracing at both light and electron microscopic levels. We have developed and employed these combined approaches to study the mechanisms underlying the development and refinement of climbing fiber mono-innervation in cerebellar Purkinje cells . Therefore, we present some examples of the images obtained in this experimental context.

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Authors and Affiliations

  1. Department of Anatomy, Hokkaido University School of Medicine, Kita 15 Nishi 7, Kita-ku, Sapporo, 060-8638, Japan

    Taisuke Miyazaki & Masahiko Watanabe

Authors
  1. Taisuke Miyazaki
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  2. Masahiko Watanabe
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Corresponding authors

Correspondence to Taisuke Miyazaki or Masahiko Watanabe .

Editor information

Editors and Affiliations

  1. Department of Veterinary Sciences, University of Torino, Grugliasco, Torino, Torino, Italy

    Adalberto Merighi

  2. Department of Veterinary Sciences, University of Torino, Grugliasco, Torino, Torino, Italy

    Laura Lossi

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Miyazaki, T., Watanabe, M. (2015). Combined Immunocytochemistry and Tracing of Neural Connections. In: Merighi, A., Lossi, L. (eds) Immunocytochemistry and Related Techniques. Neuromethods, vol 101. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2313-7_16

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  • DOI: https://doi.org/10.1007/978-1-4939-2313-7_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2312-0

  • Online ISBN: 978-1-4939-2313-7

  • eBook Packages: Springer Protocols

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