A bovine herpesvirus type 1 mutant virus with truncated glycoprotein E cytoplasmic tail has defective anterograde neuronal transport in rabbit dorsal root ganglia primary neuronal cultures in a microfluidic chamber system
- 85 Downloads
Bovine herpesvirus type 1 (BHV-1) is an important component of the bovine respiratory disease complex (BRDC) in cattle. Following primary intranasal and ocular infection of cattle, BHV-1 establishes lifelong latent infection in trigeminal ganglia (TG). Upon reactivation from latency, the virus is transported from neuronal cell bodies in the TG to projected nerve endings in nose and cornea of latently infected cattle where the virus shedding occurs. This property of BHV-1 plays a significant role in the pathogenesis of BRDC and maintenance of BHV-1 in the cattle population. Recently, we have reported that a glycoprotein E (gE) cytoplasmic tail-truncated BHV-1 (BHV-1 gEAm453) did not reactivate from latency and was not shed in the nasal and ocular secretions of calves and rabbits. Here we describe the methods to establish rabbit primary dorsal root ganglia (DRG) neuron cultures in a microfluidic chamber system and to characterizein vitro anterograde and retrograde axonal transport properties of BHV-1 gE-deleted and BHV-1 cytoplasmic tail-truncated gEAm453 mutant viruses relative to BHV-1 gEAm453-rescued/ wild-type viruses. The results clearly demonstrated that whereas the BHV-1 gE-deleted, BHV-1 gEAm453, and BHV-1 gEAm453-rescued/wild-type viruses were transported equally efficiently in the retrograde direction, only the BHV-1 gEAm453-rescued/wild-type virus was transported anterogradely. Therefore, we have concluded that sequences within the BHV-1 gE cytoplasmic tail are essential for anterograde axonal transport and that primary rabbit DRG neuronal cultures in the microfluidic chambers are suitable for BHV-1 neuronal transport studies.
KeywordsBHV-1 enveope glycoprotein gE compartmentalized primary neuron culture defective anterograde neuronal transport gE cytoplasmic tail-truncated virus
Unable to display preview. Download preview PDF.
- Jiang Y, Hossain AM, Winkler T, Holt T, Doster A, Jones C (1998). A protein encoded by the latency-related gene of bovine herpesvirus 1 is expressed in trigeminal ganglionic neurons of latently infected cattle and interacts with cyclin-dependent kinase 2 during productive infection.J Virol 72: 8133–8142.PubMedGoogle Scholar
- Liu Z-F, Brum M, Doster A, Jones C, Chowdhury SI (2008b). A bovine herpesvirus type 1 mutant virus specifying a carboxyl-terminal truncation of glycoprotein E is defective in anterograde neuronal transport in rabbits and calves.J Virol 82: 7432–7442. Maresch C, Granzow H, Negatsch A, Klupp BG, Fuchs W,CrossRefPubMedGoogle Scholar
- Snyder A, Polcicova K, Johnson DC (2008). Herpes simplex virus gE/gI and Us9 proteins promote transport of both capsids and virion glycoproteins in neuronal axons.J NeuroVirol 82: 10613–10624.Google Scholar
- Wyler R, Engels M, Schwyzer M (1989). Infectious bovine rhinotracheitis/vulvo-vaginitis (BHV-1). In:Herpesvirus disease of cattle horses and pigs. Wittman, G (ed). Hingham, MA: Kluwer Academic Publishers, pp. 1–72.Google Scholar