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Methods and Applications of Campenot Trichamber Neuronal Cultures for the Study of Neuroinvasive Viruses

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Axonal Transport

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2431))

Abstract

The development of compartmentalized neuron culture systems has been invaluable in the study of neuroinvasive viruses, including the alpha herpesviruses Herpes Simplex Virus 1 (HSV-1) and Pseudorabies Virus (PRV). This chapter provides updated protocols for assembling and culturing rodent embryonic superior cervical ganglion (SCG) and dorsal root ganglion (DRG) neurons in Campenot trichamber cultures. In addition, we provide several illustrative examples of the types of experiments that are enabled by Campenot cultures: (1) Using fluorescence microscopy to investigate axonal outgrowth/extension through the chambers, and alpha herpesvirus infection, intracellular trafficking, and cell-cell spread via axons. (2) Using correlative fluorescence microscopy and cryo electron tomography to investigate the ultrastructure of virus particles trafficking in axons.

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Acknowledgments

We would like to especially thank Dr. Lynn Enquist, Princeton University, for his ongoing mentorship. Much of this work would not have been possible without his generosity, guidance, and patience, which directly contributed to the development of these methods. This work was supported by NIH grants K22 AI123159 (I.B.H.), R21 AI146952 (M.P.T.), P40 OD010996 (E.A.E.), and P41 GM103832 (W.C.).

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Author notes

  1. Wesley Tierney and Ian A. Vicino contributed equally to this work.

Authors and Affiliations

  1. Center for Immunotherapy, Vaccines, and Virotherapy, Biodesign Institute, and School of Life Sciences, Arizona State University, Tempe, AZ, USA

    Wesley M. Tierney, Ian A. Vicino & Ian B. Hogue

  2. Department of Bioengineering, Department of Microbiology and Immunology, Division of CryoEM and Bioimaging, SSRL, SLAC National Accelerator Laboratory, Stanford University, Menlo Park, CA, USA

    Stella Y. Sun & Wah Chiu

  3. Department of Molecular Biology and Princeton Neuroscience Institute, Princeton University, Princeton, NJ, USA

    Esteban A. Engel

  4. Department of Microbiology and Cell Biology, Montana State University, Bozeman, MT, USA

    Matthew P. Taylor

Authors
  1. Wesley M. Tierney
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  2. Ian A. Vicino
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  3. Stella Y. Sun
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  4. Wah Chiu
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  5. Esteban A. Engel
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  6. Matthew P. Taylor
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  7. Ian B. Hogue
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Corresponding authors

Correspondence to Matthew P. Taylor or Ian B. Hogue .

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Editors and Affiliations

  1. Department of Basic and Clinical Neurosciences, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King’s College London, London, UK

    Alessio Vagnoni

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Tierney, W.M. et al. (2022). Methods and Applications of Campenot Trichamber Neuronal Cultures for the Study of Neuroinvasive Viruses. In: Vagnoni, A. (eds) Axonal Transport. Methods in Molecular Biology, vol 2431. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1990-2_9

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  • DOI: https://doi.org/10.1007/978-1-0716-1990-2_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1989-6

  • Online ISBN: 978-1-0716-1990-2

  • eBook Packages: Springer Protocols

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