Immediate early genes of glucocorticoid action on the developing intestine
Immediate early genes of glucocorticoid action on the developing intestine
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DOI：
10.1152/ajpgi.00454.2004
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发表时间：
2005-05-01
影响因子：
4.5
通讯作者：
Henning, SJ
中科院分区：
文献类型：
--
作者：
Agbemafle, BM;Oesterreicher, TJ;Henning, SJ
先前的研究已经证明，糖皮质激素引起小肠的功能成熟，如它们诱导各种消化水解酶（例如蔗糖酶-异麦芽糖酶和海藻糖酶）表达增加的能力所证明的。然而，这些增加具有类似于24小时的滞后时间，这表明它们是激素作用的次级效应。为了鉴定候选的主要应答基因，我们对来自未治疗的出生后第8天的小鼠幼仔和早期接受地塞米松2小时的同窝小鼠的空肠的合并RNA进行了微阵列分析。将荧光染料标记的样品以四倍体与含有来自国家衰老研究所cDNA克隆集的15，000个cDNA克隆的玻璃斑点cDNA微阵列杂交。使用相对严格的标准分析所得到的信号确定66转录上调和36下调2小时的糖皮质激素治疗。在上调的转录本中，微阵列检测到的增加幅度为1.4至16倍。随后通过北方印迹分析分析从整个范围中选择的mRNA。在所选的11种mRNA中，所有mRNA都得到了证实，并且在从微阵列分析和从北方印迹分析观察到的增加幅度之间存在强相关性。其他时间点显示，这些转录本在2 - 6 h之间达到峰值，并在24 h时恢复至基线水平。基因本体论分析显示地塞米松对发育中的肠具有多效性作用，并指出发育类别中的基因可能是糖皮质激素诱导的肠功能成熟的调解候选基因。
Prior studies have demonstrated that glucocorticoid hormones elicit functional maturation of the small intestine as evidenced by their ability to induce increases in the expression of various digestive hydrolases, such as sucrase-isomaltase and trehalase. However, these increases have a lag time of similar to 24 h, suggesting that they are secondary effects of hormone action. To identify candidate primary response genes, we performed microarray analysis on pooled RNA from jejunums of untreated postnatal day 8 mouse pups and from littermates who earlier received dexamethasone 2 h. Fluorescent dye-labeled samples were hybridized in quadruplicate to glass-spotted cDNA microarrays containing 15,000 cDNA clones from the National Institute of Aging cDNA clone set. Analysis of the resulting signals using relatively stringent criteria identified 66 transcripts upregulated and 36 down-regulated by 2 h of glucocorticoid treatment. Among the upregulated transcripts, the magnitude of the increase detected by microarray ranged from 1.4- to 16-fold. Selected mRNAs from throughout the range were subsequently analyzed by Northern blot analysis. Of 11 mRNAs chosen all were confirmed, and there was a strong correlation between the magnitude of the increase observed from the microarray analysis and from Northern blot analysis. Additional time points showed that these transcripts peaked between 2 and 6 h and had returned to baseline by 24 h. Gene ontology analysis showed pleiotropic effects of dexamethasone on the developing intestine and pointed to genes in the development category as being likely candidates for mediation of glucocorticoid-induced maturation of intestinal function.