Bacon Andrea, Kerr Niall C H, Holmes Fiona E, Gaston Kevin, Wynick David
Department of Pharmacology, Bristol University, Bristol BS8 1TD, United Kingdom.
J Neurosci. 2007 Jun 13;27(24):6573-80. doi: 10.1523/JNEUROSCI.1596-07.2007.
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.
坐骨神经切断术后,背根神经节(DRG)中甘丙肽的表达显著增加,并调节疼痛行为和感觉神经元的再生。在此,我们描述了转基因小鼠,其表达带有由LacZ标记的鼠甘丙肽基因上游不同数量序列的构建体。甘丙肽基因上游20 kb区域概括了甘丙肽在胚胎和成年完整DRG以及切断术后的内源性表达模式。相比之下,1.9 kb的片段在完整DRG或切断术后均未能驱动LacZ表达。然而,添加额外的2.7 kb 5'侧翼DNA(4.6 kb构建体)可恢复胚胎DRG和成年切断术后的表达。对该2.7 kb区域的序列分析揭示了独特的18 bp和23 bp区域,分别包含重叠的假定Ets、Stat和Smad结合位点以及相邻的假定Stat和Smad结合位点。从4.6 kb构建体中删除18 bp和23 bp区域消除了切断术后DRG中LacZ表达的上调,但不影响胚胎或成年完整DRG中的表达。此外,对许多其他切断术反应性基因上游区域的生物信息学分析表明,假定的Ets、Stat和Smad结合位点紧密相邻似乎是损伤诱导基因表达上调中的一个共同基序。