Characterization of an enhancer region of the galanin gene that directs expression to the dorsal root ganglion and confers responsiveness to axotomy.
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.
Bacon
Andrea
A
Department of Pharmacology, Bristol University, Bristol BS8 1TD, United Kingdom.
Kerr
Niall C H
NC
Holmes
Fiona E
FE
Gaston
Kevin
K
Wynick
David
D
D000818:Animals; D019771:Axotomy / Q000379:methods; D004622:Embryo, Mammalian; D004742:Enhancer Elements, Genetic / Q000502:physiology*; D019004:Galanin / Q000235:genetics*; D005727:Ganglia, Spinal / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology*; D007763:Lac Operon / Q000502:physiology; D051379:Mice; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D009154:Mutation / Q000502:physiology; D017434:Protein Structure, Tertiary; D012584:Sciatic Nerve / Q000378:metabolism / Q000601:surgery; D017421:Sequence Analysis; D001616:beta-Galactosidase / Q000378:metabolism
D019004:Galanin; D001616:beta-Galactosidase