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Updated: Jul 6, 2026
A Primary Neuron Culture System for the Study of Herpes Simplex Virus Latency and Reactivation
Published on: April 2, 2012
Diverse stabilities of expression in the rat brain from different cellular promoters in a helper virus-free herpes
Y Wang1, L Yu, A I Geller
1Division of Endocrinology, Children's Hospital, Boston, MA 02115, USA.
Abstract:
Many neural gene transfer studies require both long-term and cell type-specific expression. We have reported a helper virus-free HSV-1 plasmid vector system (Fraefel et al., 1996), and this system supports at least some long-term expression from herpesvirus immediate-early promoters. In this study, we constructed vectors that placed the lacZ reporter gene under the regulation of five different cellular promoters. Vector stocks were microinjected into the midbrain, striatum, or hippocampus; the rats were sacrificed at 4 days to 2 months after gene transfer; and the numbers of X-Gal-positive cells were determined. A 6.8-kb fragment of the rat tyrosine hydroxylase (TH) promoter supported relatively stable expression for up to 2 months and targeted expression to TH-immunoreactive neurons in the substantia nigra pars compacta. The other promoters that were examined were chosen with the goal of obtaining long-term, neuronal-specific expression. At 4 days after gene transfer, a 766-bp fragment of the TH promoter supported expression in cells with neuronal morphology in the midbrain and striatum, consistent with results in transgenic mice. However, expression was absent by 2 weeks. Similarly, at 4 days after gene transfer, a mouse neurofilament heavy subunit promoter supported expression in cells with neuronal morphology in the midbrain, striatum, and hippocampus, but expression was absent by 2 weeks. A rat neuron-specific enolase promoter supported only a low level of expression in cultured neuronal cells, and expression was not detected in the brain. A rat voltage-gated sodium channel promoter supported only a low level of expression in PC12 cells and expression could not be detected in cultured cortical cells. These results demonstrate that different promoters support a wide range of levels and stabilities of expression in this vector system, and the results suggest approaches to improving the stability of long-term expression.
