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首页> 外文期刊>American Journal of Physiology >Rapid induction of GATA transcription factors in developing mouse intestine following glucocorticoid administration.
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Rapid induction of GATA transcription factors in developing mouse intestine following glucocorticoid administration.

机译：糖皮质激素给药后，在发育中的小鼠肠道中快速诱导GATA转录因子。

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In the developing intestine, transcription of alpha-glucosidase genes such as sucrase-isomaltase and trehalase is stimulated by glucocorticoid administration. The consequent increase of their respective mRNAs is characterized by a 12-h lag, suggesting that the response to glucocorticoids represents a secondary effect. We hypothesized that the primary response of the tissue to glucocorticoids includes induction of one or more intestinal transcription factors. To investigate this hypothesis, we identified a region in the mouse trehalase promoter (located at nucleotides -406 to -377 from the transcription start site) with potential binding sites for three transcription factors: Cdx-2, GATA, and C/EBP. Gel shifts were performed using labeled oligonucleotides from this region with nuclear extracts from jejunums of either control 8-day-old mouse pups or littermates treated with dexamethasone (DEX) 4 h before death. A specific shifted band was observed with DEX extracts but not with control extracts. Supershift assays indicated the presence of GATA-4 and GATA-6 but not GATA-5 nor Cdx-2, C/EBP alpha, C/EBP beta, or C/EBP delta. GATA binding was further implicated by competition studies with mutated oligonucleotides. Finally, Western blot analysis showed GATA-4 and GATA-6 proteins in DEX but not control nuclear extracts. For GATA-4, the same pattern was demonstrated with whole cell extracts and with the cytosol fraction. We conclude that expression of GATA-4 and GATA-6 proteins in the suckling mouse jejunum is stimulated by DEX. This novel finding constitutes an important first step in understanding the molecular mechanism of glucocorticoid action on the developing intestine.
机译：在发育中的肠中，通过糖皮质激素的给药刺激α-葡糖苷酶基因例如蔗糖酶-异麦芽糖酶和海藻糖酶的转录。其相应的mRNA随之增加的特征是滞后12小时，这表明对糖皮质激素的反应代表了次级作用。我们假设组织对糖皮质激素的主要反应包括诱导一种或多种肠道转录因子。要研究此假设，我们在小鼠海藻糖酶启动子中确定了一个区域（位于转录起始位点的核苷酸-406至-377），具有三个转录因子（Cdx-2，GATA和C / EBP）的潜在结合位点。使用来自该区域的标记的寡核苷酸和死前4小时用地塞米松（DEX）处理的对照8日龄小鼠幼仔或同窝仔的空肠核提取物进行凝胶转移。用DEX提取物观察到特定的移位带，而用对照提取物观察不到。 Supershift分析表明存在GATA-4和GATA-6，但不存在GATA-5或Cdx-2，C / EBP alpha，C / EBP beta或C / EBP delta。通过与突变寡核苷酸的竞争研究进一步暗示了GATA结合。最后，蛋白质印迹分析显示了DEX中的GATA-4和GATA-6蛋白，但没有对照核提取物。对于GATA-4，全细胞提取物和细胞溶质组分也显示出相同的模式。我们得出结论，DEX刺激了乳鼠空肠中GATA-4和GATA-6蛋白的表达。这一新发现是理解糖皮质激素作用于发育中小肠的分子机制的重要第一步。

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