MicroRNA expression is required for pancreatic islet cell genesis in the mouse
MicroRNA expression is required for pancreatic islet cell genesis in the mouse
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DOI：
10.2337/db07-0175
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发表时间：
2007-12-01
期刊：
影响因子：
7.7
通讯作者：
German, Michael S.
中科院分区：
文献类型：
--
作者：
Lynn, Francis C.;Skewes-Cox, Peter;German, Michael S.
胰腺发育过程中不同细胞类型的产生取决于基因表达的顺序变化。我们测试的假设，即microRNAs（miRNA），限制基因表达，通过转录后沉默，调节参与胰腺development.RESEARCH设计和方法的基因表达级联克隆和测序从发展中的胰腺，这些基因的一个子集的表达进行了测试，使用锁核酸原位分析。为了评估miRNA对胰腺发育的总体贡献，从发育中的胰腺中有条件地删除了Dicer 1（一种miRNA加工所需的酶）JETS-小RNA测序鉴定了超过125种miRNA，包括18种新序列，其在发育中的胰腺中具有不同的表达结构域。为了测试这些miRNA对发育的贡献，我们在胰腺发育的早期有条件地删除了miRNA加工酶Dicer 1。Dicer无效的动物在所有胰腺谱系中显示出严重缺陷，尽管内分泌细胞，尤其是产生胰岛素的β细胞减少得最显著。内分泌缺陷与notch信号靶点Hes 1的增加和表达Hes 1靶基因neurogenin 3的内分泌祖细胞形成的减少有关。结论-在胰腺发育过程中需要表达独特的miRNA谱，并且是β细胞形成所必需的。
OBJECTIVE-The generation of distinct cell types during the development of the pancreas depends on sequential changes in gene expression. We tested the hypothesis that microRNAs (miRNAs), which limit gene expression through posttranscriptional silencing, modulate the gene expression cascades involved in pancreas development.RESEARCH DESIGN AND METHODS-miRNAs were cloned and sequenced from developing pancreata, and expression of a subset of these genes was tested using locked nucleic acid in situ analyses. To assess the overall contribution of miRNAs to pancreatic development, Dicerl, an enzyme required for miRNA processing, was conditionally deleted from the developing pancreas.RESULTS-Sequencing of small RNAs identified over 125 miRNAs, including 18 novel sequences, with distinct expression domains within the developing pancreas. To test the developmental contribution of these miRNAs, we conditionally deleted the miRNA processing enzyme Dicer1 early in pancreas development. Dicer-null animals displayed gross defects in all pancreatic lineages, although the endocrine cells, and especially the insulin-producing beta-cells, were most dramatically reduced. The endocrine defect was associated with an increase in the notch-signaling target Hes1 and a reduction in the formation of endocrine cell progenitors expressing the Hes1 target gene neurogenin3.CONCLUSIONS-The expression of a unique profile of miRNAs is required during pancreas development and is necessary for beta-cell formation.