title: 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.
abstract: 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 characterize in 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.
authors.affiliation: Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana 70803, USA. Chowdh@lsu.edu
authors.forename: S I
authors.initials: SI
authors.lastname: Chowdhury
authors.forename: J
authors.initials: J
authors.lastname: Coats
authors.forename: R A
authors.initials: RA
authors.lastname: Neis
authors.forename: S M
authors.initials: SM
authors.lastname: Navarro
authors.forename: D B
authors.initials: DB
authors.lastname: Paulsen
authors.forename: J-M
authors.initials: JM
authors.lastname: Feng
mesh_terms: D000818:Animals; D048090:Bovine Respiratory Disease Complex / Q000821:virology; D002417:Cattle; D002418:Cattle Diseases / Q000821:virology; D002478:Cells, Cultured; D004285:Dogs; D005260:Female; D005727:Ganglia, Spinal / Q000378:metabolism / Q000821:virology*; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D007242:Herpesvirus 1, Bovine / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000821:virology*; D011817:Rabbits; D012668:Trigeminal Ganglion / Q000821:virology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D017376:Viral Tail Proteins / Q000378:metabolism*; D014775:Virus Activation / Q000235:genetics; D017735:Virus Latency / Q000235:genetics
chemical_list: D006023:Glycoproteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D017376:Viral Tail Proteins; C046998:bovine herpesvirus type-1 glycoproteins