title: In vitro analysis of transneuronal spread of an alphaherpesvirus infection in peripheral nervous system neurons.
abstract: The neurotropic alphaherpesviruses invade and spread in the nervous system in a directional manner between synaptically connected neurons. Until now, this property has been studied only in living animals and has not been accessible to in vitro analysis. In this study, we describe an in vitro system in which cultured peripheral nervous system neurons are separated from their neuron targets by an isolator chamber ring. Using pseudorabies virus (PRV), an alphaherpesvirus capable of transneuronal spread in neural circuits of many animals, we have recapitulated in vitro all known genetic requirements for retrograde and anterograde transneuronal spread as determined previously in vivo. We show that in vitro transneuronal spread requires intact axons and the presence of the viral proteins gE, gI, and Us9. We also show that transneuronal spread is dependent on the viral glycoprotein gB, which is required for membrane fusion, but not on gD, which is required for extracellular spread. We demonstrate ultrastructural differences between anterograde- and retrograde-traveling virions. Finally, we show live imaging of dynamic fluorescent virion components in axons and postsynaptic target neurons.
authors: 301 Schultz Building, Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA. becket@princeton.edu
authors: B
authors: B
authors: Feierbach
authors: M
authors: M
authors: Bisher
authors: J
authors: J
authors: Goodhouse
authors: L W
authors: LW
authors: Enquist
mesh_terms: D000818:Animals; D001369:Axons / Q000648:ultrastructure / Q000821:virology*; D001692:Biological Transport / Q000235:genetics; D002417:Cattle; D002460:Cell Line; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D010525:Peripheral Nerves / Q000378:metabolism / Q000821:virology*; D013552:Swine; D013569:Synapses / Q000235:genetics / Q000648:ultrastructure / Q000821:virology*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D014759:Viral Envelope Proteins