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Journal of Virology, October 1999, p. 8503-8511, Vol. 73, No. 10
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Anterograde Transport of Herpes Simplex Virus
Proteins in Axons of Peripheral Human Fetal Neurons: an
Immunoelectron Microscopy Study
David J.
Holland,1
Monica
Miranda-Saksena,1
Ross A.
Boadle,1,2
Patricia
Armati,3 and
Anthony
L.
Cunningham1,*
Centre for Virus Research, Westmead
Institutes of Health Research, Westmead Hospital and University of
Sydney,1 and Westmead Institutes of
Health Research and Electron Microscope Laboratory, Westmead
Hospital,2 Westmead, New South Wales 2145, and School of Biological Sciences, University of Sydney,
New South Wales 2006,3 Australia
Received 30 April 1999/Accepted 8 July 1999
Herpes simplex virus (HSV) reactivates from latency in the neurons
of dorsal root ganglia (DRG) and is subsequently transported anterogradely along the axon to be shed at the skin or mucosa. Although
we have previously shown that only unenveloped nucleocapsids are
present in axons during anterograde transport, the mode of transport of
tegument proteins and glycoproteins is not known. We used a two-chamber
culture model with human fetal DRG cultivated in an inner chamber,
allowing axons to grow out and penetrate an agarose barrier and
interact with autologous epidermal cells in the outer chamber. After
HSV infection of the DRG, anterograde transport of viral components
could be examined in the axons in the outer chamber at different time
points by electron and immunoelectron microscopy (IEM). In the axons,
unenveloped nucleocapsids or focal collections of gold immunolabel for
nucleocapsid (VP5) and/or tegument (VP16) were detected. VP5 and VP16
usually colocalized in both scanning and transmission IEM. In contrast,
immunolabel for glycoproteins gB, gC, and gD was diffusely
distributed in axons and was rarely associated with VP5 or VP16. In
longitudinal sections of axons, immunolabel for glycoprotein was
arrayed along the membranes of axonal vesicles. These findings provide
evidence that in DRG axons, virus nucleocapsids coated with tegument
proteins are transported separately from glycoproteins and suggest that final assembly of enveloped virus occurs at the axon terminus.
*
Corresponding author. Mailing address: Centre for Virus
Research, Westmead Institutes of Health Research, Westmead Hospital and
University of Sydney, Westmead, New South Wales 2145, Australia. Phone:
(61) 2 9845 6229. Fax: (61) 2 9845 8300. E-mail:
tonyc{at}westgate.wh.usyd.edu.au.
Journal of Virology, October 1999, p. 8503-8511, Vol. 73, No. 10
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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