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Journal of NeuroVirology

, Volume 8, Issue 3, pp 204–210 | Cite as

The relationship of herpes simplex virus latency associated transcript expression to genome copy number: A quantitative study using laser capture microdissection

  • Xiao-Ping Chen
  • Marina Mata
  • Mary Kelley
  • Joseph C. Glorioso
  • David J. FinkEmail author
Article

Abstract

To investigate the quantitative relationship of latent herpes simplex virus (HSV) genomes to the expression of latency associated transcripts (LATs) we used a combination of laser capture microdissection (LCM), polymerase chain reaction (PCR), and quantitative real-time PCR to determine the number of HSV genomes in individual neurons of the mouse trigeminal ganglion (TG) during viral latency. Both LAT-positive and LAT-negative neurons detected by in situ hybridization (ISH) and lifted by LCM contained HSV genomes detected by PCR for HSV ICP47. The number of genomes/cell determined by real-time PCR with probes for HSV UL44 following LCM demonstrated a Poisson distribution with a predicted mean count of 178 genomes /LAT-positive neuron, and 68 genomes/LAT-negative neuron. The range was similar between the LAT-positive and LAT-negative neurons, and there was a substantial overlap in the distributions. These results suggest that the expression of LATs in an amount that is detectable by ISH does not depend only on the number of HSV genomes in the cell, and by implication suggests that neuron-specific factors play a role in the regulation of LAT expression during latency.

Keywords

herpes simplex virus latency latency associated transcript (LAT) real-time PCR laser capture microdissection 

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Copyright information

© Journal of NeuroVirology, Inc. 2002

Authors and Affiliations

  • Xiao-Ping Chen
    • 1
  • Marina Mata
    • 1
    • 3
  • Mary Kelley
    • 3
  • Joseph C. Glorioso
    • 2
  • David J. Fink
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of NeurologyUniversity of Pittsburgh School of MedicinePittsburgh
  2. 2.Department of Molecular Genetics and BiochemistryUniversity of Pittsburgh School of MedicinePittsburgh
  3. 3.VA Medical CenterGRECCPittsburghUSA

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