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The Journal of Neuroscience, June 13, 2007, 27(24):6573-6580; doi:10.1523/JNEUROSCI.1596-07.2007

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Cellular/Molecular
Characterization of an Enhancer Region of the Galanin Gene That Directs Expression to the Dorsal Root Ganglion and Confers Responsiveness to Axotomy

Andrea Bacon,¹ Niall C. H. Kerr,¹ Fiona E. Holmes,¹ Kevin Gaston,² and David Wynick¹

¹Departments of Pharmacology and Clinical Sciences South Bristol and ²Department of Biochemistry, School of Medical Sciences, University Walk, Bristol University, Bristol BS8 1TD, United Kingdom

Correspondence should be addressed to Dr. David Wynick, School of Medical Sciences, University Walk, University of Bristol, Bristol BS8 1TD, UK. Email: d.wynick{at}bris.ac.uk

Galanin expression markedly increases in the dorsal root ganglion (DRG) after sciatic nerve axotomy and modulates pain behavior and regeneration of sensory neurons. Here, we describe transgenic mice expressing constructs with varying amounts of sequence upstream of the murine galanin gene marked by LacZ. The 20 kb region upstream of the galanin gene recapitulates the endogenous expression pattern of galanin in the embryonic and adult intact DRG and after axotomy. In contrast, 1.9 kb failed to drive LacZ expression in the intact DRG or after axotomy. However, the addition of an additional 2.7 kb of 5' flanking DNA (4.6 kb construct) restored the expression in the embryonic DRG and in the adult after axotomy. Sequence analysis of this 2.7 kb region revealed unique 18 and 23 bp regions containing overlapping putative Ets-, Stat-, and Smad-binding sites, and adjacent putative Stat- and Smad-binding sites, respectively. Deletion of the 18 and 23 bp regions from the 4.6 kb construct abolished the upregulation of LacZ expression in the DRG after axotomy but did not affect expression in the embryonic or intact adult DRG. Also, a bioinformatic analysis of the upstream regions of a number of other axotomy-responsive genes demonstrated that the close proximity of putative Ets-, Stat-, and Smad-binding sites appears to be a common motif in injury-induced upregulation in gene expression.

Key words: galanin; DRG; Ets; Stat; Smad; axotomy

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Received March 1, 2007; revised May 14, 2007; accepted May 14, 2007.

Correspondence should be addressed to Dr. David Wynick, School of Medical Sciences, University Walk, University of Bristol, Bristol BS8 1TD, UK. Email: d.wynick{at}bris.ac.uk

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