Preprint Review Version 1 Preserved in Portico This version is not peer-reviewed

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

Version 1 : Received: 4 May 2021 / Approved: 6 May 2021 / Online: 6 May 2021 (15:08:12 CEST)

How to cite: Tierney, W.; Vicino, I.; Sun, S.; Chiu, W.; Engel, E.; Taylor, M.; Hogue, I. Methods and Applications of Campenot Trichamber Neuronal Cultures for the Study of Neuroinvasive Viruses. Preprints 2021, 2021050098. https://doi.org/10.20944/preprints202105.0098.v1 Tierney, W.; Vicino, I.; Sun, S.; Chiu, W.; Engel, E.; Taylor, M.; Hogue, I. Methods and Applications of Campenot Trichamber Neuronal Cultures for the Study of Neuroinvasive Viruses. Preprints 2021, 2021050098. https://doi.org/10.20944/preprints202105.0098.v1

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

Campenot, neurons, superior cervical ganglia, dorsal root ganglia, virus, alphaherpesvirus, herpes simplex virus, pseudorabies virus, fluorescence microscopy, cryo electron tomography

Subject

Biology and Life Sciences, Virology

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