Construction of recombinant pseudorabies viruses optimized for labeling and neurochemical characterization of neural circuitry

Zsolt Boldogköi, Aniko Reichart, Ida E. Tóth, Attila Sik, Ferenc Erdélyi, Istvan Medveczky, Catherine Llorens-Cortes, Miklos Palkovits, Zsolt Lenkei

Research output: Article

28 Citations (Scopus)

Abstract

In this study we have modified the neuroinvasiveness of pseudorabies virus strain Bartha, a commonly utilized trans-synaptic tract-tracer. In addition, we sought to facilitate detection of cellular mRNAs in neurons infected with the virus. In order to modify spreading characteristics, we inserted the lacZ or the GFP (green fluorescent protein) genes into the genomic loci containing the putative latency-associated transcript promoter (PLAT2), resulting in the disruption of the promoter function. Following rat kidney injection, mutant viruses labeled central autonomic neurons in a slower and much more restricted manner than the parent Bartha strain. Since both reporter genes were controlled by the human cytomegalovirus immediate early (IE) 1 promoter, they exhibited IE expression kinetics. This property proved to be important for the co-detection of reporter proteins with neuronal mRNAs, readily detected at early but not at late stage of infection, as shown in tyrosine-hydroxylase expressing A5 catecholaminergic neurons and in serotonin transporter expressing raphe magnus neurons.

Original languageEnglish
Pages (from-to)105-118
Number of pages14
JournalMolecular Brain Research
Volume109
Issue number1-2
DOIs
Publication statusPublished - dec. 30 2002

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience

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