Retrograde axonal transport of VZV: kinetic studies in hESC-derived neurons.
Retrograde axonal transport of the neurotropic alphaherpesvirus Varicella zoster virus (VZV) from vesicles at the skin results in sensory neuron infection and establishment of latency. Reactivation from latency leads to painful herpes zoster. The lack of a suitable animal model of these processes for the highly human-restricted VZV has resulted in a dearth of knowledge regarding the axonal transport of VZV. We recently demonstrated VZV infection of distal axons, leading to subsequent capsid transport to the neuronal somata, and replication and release of infectious virus using a new model based on neurons derived from human embryonic stem cells (hESC). In the present study, we perform a kinetic analysis of the retrograde transport of green fluorescent protein-tagged ORF23 in VZV capsids using hESC-derived neurons compartmentalized microfluidic chambers and time-lapse video microscopy. The motion of the VZV was discontinuous, showing net retrograde movement with numerous short pauses and reversals in direction. Velocities measured were higher 1 h after infection than 6 h after infection, while run lengths were similar at both time points. The hESC-derived neuron model was also used to show that reduced neuronal spread by a VZV loss-of-function mutant for ORF7 is not due to the prevention of axonal infection and transport of the virus to the neuronal somata. hESC-derived neurons are, therefore, a powerful model for studying axonal transport of VZV and molecular characteristics of neuronal infection.
Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Gonda Building, Old Campus, 52900, Ramat-Gan, Israel.
Sergei
S
Grigoryan
Paul R
PR
Kinchington
In Hong
IH
Yang
Anca
A
Selariu
Hua
H
Zhu
Michael
M
Yee
Ronald S
RS
Goldstein
D001370:Axonal Transport; D002213:Capsid / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D014645:Herpesvirus 3, Human / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D046210:Microfluidic Analytical Techniques; D008954:Models, Biological; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure / Q000821:virology*; D016366:Open Reading Frames; D059008:Time-Lapse Imaging; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D017735:Virus Latency; D014779:Virus Replication
D014764:Viral Proteins; D049452:Green Fluorescent Proteins