Compartmentalized Neuronal Culture for Viral Transport Research

Abstract

Neuron-invading viruses usually enter via the peripheral organs/tissues of their mammalian hosts and are transported to the neurons. Virus trafficking is critical for transport or spread within the nervous system. Primary culture of neurons is a valuable and indispensable method for neurobiological research, allowing researchers to investigate basic mechanisms of diverse neuronal functions as well as retrograde and anterograde virus transport in neuronal axons. Primary ganglion sensory neurons from mice can be cultured in a compartmentalized culture device, which allows spatial fluidic separation of cell bodies and distal axons. These neurons serve as an important model for investigating the transport of viruses between the neuronal soma and distal axons. Alphaherpesviruses are fascinating and important human and animal pathogens, they replicate and establish lifelong latent infection in the peripheral nervous system, the mechanism of the viral transport along the axon is the key to understand the virus spread in the nervous system. In this review, we briefly introduce and evaluate the most frequently used compartmentalization tools in viral transport research, with particular emphasis on alphaherpesviruses.

Advances in the immunoescape mechanisms exploited by alphaherpesviruses

“…In a study by Smith, freshly produced green fluorescent protein (GFP)-tagged capsids were observed to traverse monomeric red fluorescent protein-tagged capsids in opposite directions within the same axon ( Smith et al, 2004 ). Therefore, the regulation of transport direction and the resulting coordination of movement occur at the level of individual capsids rather than through the overarching control of axonal transport ( Wang et al, 2020 ).…”

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

“…In the alpha herpesvirus literature, this problem has led to great confusion over the relationship between finalassembly/envelopment and axonal transport of virus particles [16]: do non-enveloped particles in axons represent progeny particles that are not yet enveloped, or inoculum particles that have already shed their envelopes during entry? Campenot trichambers, like related microfluidics devices [17][18][19], are an important tool to clarify this problem of distinguishing virus inoculum and viral progeny (see Note 1). These compartmented neuron culture devices allow SCG and DRG neurons to extend axons between chambers, while maintaining fluidic separation between the chambers [20].…”