RRC ID 2479
Author Morris R, Morgan BS, Lewis TM, Pierce KD, Pisano A, Schofield PR.
Title In vivo somatic delivery of plasmid DNA and retrograde transport to obtain cell-specific gene expression in the central nervous system.
Journal J. Neurochem.
Abstract We utilised the retrograde transport machinery of neurones to deliver naked plasmid DNA into the central nervous system. A 5.4-kb fragment of the glycine receptor (GlyR) alpha1 subunit gene was cloned and used to drive the expression of a construct encoding for the enhanced green fluorescent protein (EGFP). Injections of the plasmid DNA in the tongue of mice resulted in the expression of the marker protein in hypoglossal motor neurones, showing that the GlyRalpha1 promoter sequence is sufficient to drive expression of the transgene. In order to determine the specificity of expression of the 5.4-kb fragment of the GlyR alpha1 subunit gene promoter, we subsequently injected the plasmid DNA into the mouse central nucleus of the amygdala. This nucleus receives projections from the parabrachial nucleus, a brainstem area that has a high density of GlyRs, and from the insular cortex, a forebrain structure devoid of GlyRs. We observed EGFP-labelled neurones in the parabrachial nucleus, but not in the insular cortex, indicating that the 5.4-kb GlyR alpha1 subunit gene promoter confers specificity of expression. This approach provides a simple and rapid way to identify, in vivo, promoter elements that mediate neurone-specific gene expression.
Volume 90(6)
Pages 1445-52
Published 2004-9
DOI 10.1111/j.1471-4159.2004.02612.x
PII JNC2612
PMID 15341528
MeSH Animals Biological Transport / physiology Cell Line Central Nervous System / cytology* Central Nervous System / metabolism Cloning, Molecular / methods DNA / metabolism* Embryo, Mammalian Gene Expression / physiology Gene Transfer Techniques Genetic Vectors / metabolism Green Fluorescent Proteins Histocytochemistry / methods Humans Luminescent Proteins / genetics Luminescent Proteins / metabolism Male Mice Mice, Inbred C57BL Neurons / metabolism* Phosphopyruvate Hydratase / genetics Phosphopyruvate Hydratase / metabolism Plasmids / genetics Promoter Regions, Genetic / physiology Receptors, Glycine / genetics Receptors, Glycine / metabolism* Tongue / innervation Tongue / metabolism Transcription, Genetic / physiology*
IF 4.083
Times Cited 2
WOS Category NEUROSCIENCES BIOCHEMISTRY & MOLECULAR BIOLOGY
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