title: Live cell imaging of alphaherpes virus anterograde transport and spread.
abstract: Advances in live cell fluorescence microscopy techniques, as well as the construction of recombinant viral strains that express fluorescent fusion proteins have enabled real-time visualization of transport and spread of alphaherpes virus infection of neurons. The utility of novel fluorescent fusion proteins to viral membrane, tegument, and capsids, in conjunction with live cell imaging, identified viral particle assemblies undergoing transport within axons. Similar tools have been successfully employed for analyses of cell-cell spread of viral particles to quantify the number and diversity of virions transmitted between cells. Importantly, the techniques of live cell imaging of anterograde transport and spread produce a wealth of information including particle transport velocities, distributions of particles, and temporal analyses of protein localization. Alongside classical viral genetic techniques, these methodologies have provided critical insights into important mechanistic questions. In this article we describe in detail the imaging methods that were developed to answer basic questions of alphaherpes virus transport and spread.
authors: Department of Immunology and Infectious Diseases, Montana State University, USA. mptaylor@montana.edu
authors: Matthew P
authors: MP
authors: Taylor
authors: Radomir
authors: R
authors: Kratchmarov
authors: Lynn W
authors: LW
authors: Enquist
mesh_terms: D018138:Alphaherpesvirinae; D000818:Animals; D006566:Herpesviridae Infections / Q000821:virology*; D008856:Microscopy, Fluorescence / Q000379:methods*; D051381:Rats; D017783:Superior Cervical Ganglion / Q000166:cytology / Q000821:virology
references: J Virol. 2006 Apr;80(7):3592-606
references: 16537627
references: Nat Protoc. 2009;4(12):1869-87
references: 20010935
references: Nat Methods. 2005 Dec;2(12):905-9
references: 16299475
references: J Vis Exp. 2009 Jan 30;(23):
references: 19229183
references: J Virol. 2006 Nov;80(22):11235-40
references: 16971439
references: J Cell Biol. 2001 Aug 20;154(4):741-52
references: 11502759
references: mBio. 2012 May 02;3(2):
references: 22448044
references: J Virol. 2005 Sep;79(17):10875-89
references: 16103140
references: Trends Cell Biol. 2009 Nov;19(11):649-55
references: 19819147
references: Proc Natl Acad Sci U S A. 2001 Mar 13;98(6):3466-70
references: 11248101
references: Methods Mol Biol. 2005;292:299-316
references: 15507717
references: mBio. 2011 Dec 20;2(6):
references: 22186611
references: Proc Natl Acad Sci U S A. 2012 Oct 16;109(42):17046-51
references: 23027939
references: Curr Protoc Cell Biol. 2009 Jun;Chapter 26:Unit 26.4
references: 19499506
references: J Virol. 2005 Apr;79(7):3903-19
references: 15767393
references: J Cell Sci. 2009 Mar 15;122(Pt 6):753-67
references: 19261845
references: Cold Spring Harb Protoc. 2012 Mar 01;2012(3):340-3
references: 22383653
references: J Virol. 2008 Nov;82(21):10613-24
references: 18753205
references: J Virol. 2006 Nov;80(22):11165-77
references: 16971450
references: Nat Methods. 2012 Jan 30;9(2):152-8
references: 22290187
references: Cell Struct Funct. 2002 Oct;27(5):335-41
references: 12502887
references: Nat Rev Microbiol. 2011 May;9(5):382-94
references: 21494278
references: Cell Host Microbe. 2012 May 17;11(5):504-14
references: 22607803
references: Curr Opin Microbiol. 2006 Aug;9(4):423-9
references: 16814597
references: Cell Host Microbe. 2012 Dec 13;12(6):806-14
references: 23245325
references: Rev Med Virol. 2012 Nov;22(6):378-91
references: 22807192