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

Wesley M Tierney; Ian A Vicino; Stella Y Sun; Wah Chiu; Esteban A Engel; Matthew P Taylor; Ian B Hogue

doi:10.1007/978-1-0716-1990-2_9

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

Methods Mol Biol. 2022:2431:181-206. doi: 10.1007/978-1-0716-1990-2_9.

Authors

Wesley M Tierney^#¹, Ian A Vicino^#¹, Stella Y Sun², Wah Chiu², Esteban A Engel³, Matthew P Taylor⁴, Ian B Hogue⁵

Affiliations

¹ Center for Immunotherapy, Vaccines, and Virotherapy, Biodesign Institute, and School of Life Sciences, Arizona State University, Tempe, AZ, USA.
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.
³ Department of Molecular Biology and Princeton Neuroscience Institute, Princeton University, Princeton, NJ, USA.
⁴ Department of Microbiology and Cell Biology, Montana State University, Bozeman, MT, USA. mptaylor@montana.edu.
⁵ Center for Immunotherapy, Vaccines, and Virotherapy, Biodesign Institute, and School of Life Sciences, Arizona State University, Tempe, AZ, USA. ihogue@asu.edu.

^# Contributed equally.

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.

Keywords: Alphaherpesvirus; Campenot; Cryo electron tomography; Dorsal root ganglia; Fluorescence microscopy; Herpes simplex virus; Neurons; Pseudorabies virus; Superior cervical ganglia; Virus.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Axonal Transport / physiology
Axons / metabolism
Herpesvirus 1, Human* / physiology
Herpesvirus 1, Suid*
Neurons

Abstract

Publication types

MeSH terms

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