Detection of herpes simplex virus type 1 latency-associated transcript expression in trigeminal ganglia by in situ reverse transcriptase PCR.
One of the defining characteristics of herpes simplex virus type 1 (HSV-1) infection is the ability of the virus to establish a lifelong latent state in neurons. We previously demonstrated (R. Ramakrishnan, A.J. Fink, G. Jiang, P. Desai, J. C. Glorioso, and M. Levine, J. Virol. 68:1864-1873, 1994) by in situ PCR that many more neurons contain viral genomes than are detected by in situ hybridization for HSV latency-associated transcripts (LATs). To determine whether all cells which contain genomes express LATs, we examined trigeminal ganglia for LATs 1 and 8 weeks after corneal scarification with ribonucleotide reductase-deficient HSV-1 by in situ reverse transcriptase PCR. The number of LAT-positive cells detected by in situ reverse transcriptase was substantially greater than the number of cells positive by in situ hybridization and appeared to be similar to the number of cells containing HSV genomes by in situ PCR and the number of ganglionic neurons that project to the cornea as detected by retrograde labeling with Fluorogold. These results demonstrate LAT expression in many neurons containing HSV-1 genomes.
Ramakrishnan
R
R
Department of Neurology, University of Pittsburgh, Pennsylvania 15261, USA.
Poliani
P L
PL
Levine
M
M
Glorioso
J C
JC
Fink
D J
DJ
D000818:Animals; D001483:Base Sequence; D003315:Cornea / Q000473:pathology / Q000821:virology; D017931:DNA Primers; D006561:Herpes Simplex / Q000473:pathology / Q000503:physiopathology; D017403:In Situ Hybridization; D008297:Male; D008969:Molecular Sequence Data; D009474:Neurons / Q000473:pathology / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012194:RNA-Directed DNA Polymerase; D051381:Rats; D017207:Rats, Sprague-Dawley; D012264:Ribonucleotide Reductases / Q000235:genetics; D018139:Simplexvirus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D013997:Time Factors; D014158:Transcription, Genetic; D012668:Trigeminal Ganglion / Q000473:pathology / Q000821:virology*; D017735:Virus Latency
D017931:DNA Primers; D012264:Ribonucleotide Reductases; D012194:RNA-Directed DNA Polymerase