Anatomy and Cell Biology

Korean Association of Anatomists (대한해부학회)
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Evaluation of recombinant adenovirusmediated gene delivery for expression of tracer genes in catecholaminergic neurons

  • Kim, Mi-La (Department of Anatomy and Neurobiology, School of Medicine, Kyung Hee University) ;
  • Han, Sheng-Jun (Department of Anatomy and Neurobiology, School of Medicine, Kyung Hee University) ;
  • Lee, Sat-Byol (Department of Anatomy and Neurobiology, School of Medicine, Kyung Hee University) ;
  • Kim, Jung-Hye (Department of Anatomy and Neurobiology, School of Medicine, Kyung Hee University) ;
  • Ahn, Hee-Kyung (Department of Anatomy and Neurobiology, School of Medicine, Kyung Hee University) ;
  • Huh, Young-Buhm (Department of Anatomy and Neurobiology, School of Medicine, Kyung Hee University)
  • Received : 2010.05.14
  • Accepted : 2010.06.08
  • Published : 2010.06.30
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Abstract

Selective labeling of small populations of neurons of a given phenotype for conventional neuronal tracing is difficult because tracers can be taken up by all neurons at the injection site, resulting in nonspecific labeling of unrelated pathways. To overcome these problems, genetic approaches have been developed that introduce tracer proteins as transgenes under the control of cell-type-specific promoter elements for visualization of specific neuronal pathways. The aim of this study was to explore the use of tracer gene expression for neuroanatomical tracing to chart the complex interconnections of the central nervous system. Genetic tracing methods allow for expression of tracer molecules using cell-type-specific promoters to facilitate neuronal tracing. In this study, the rat tyrosine hydroxylase (TH) promoter and an adenoviral delivery system were used to express tracers specifically in dopaminergic and noradrenergic neurons. Region-specific expression of the transgenes was then analyzed. Initially, we characterized cell-type-specifi c expression of GFP or RFP in cultured cell lines. We then injected an adenovirus carrying the tracer transgene into several brain regions using a stereotaxic apparatus. Three days aft er injection, strong GFP expression was observed in the injected site of the brain. RFP and WGA were expressed in a cell-type-specific manner in the cerebellum, locus coeruleus, and ventral tegmental regions. Our results demonstrate that selective tracing of catecholaminergic neuronal circuits is possible in the rat brain using the TH promoter and adenoviral expression.

Keywords

Acknowledgement

Supported by : Kyung Hee University Research Fund

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