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通过转录组分析揭示黑腹果蝇早期胚胎中Dpp/BMP信号通路的靶基因。

Target genes of Dpp/BMP signaling pathway revealed by transcriptome profiling in the early D.melanogaster embryo.

作者信息

Dominguez Calixto, Zuñiga Alejandro, Hanna Patricia, Hodar Christian, Gonzalez Mauricio, Cambiazo Verónica

机构信息

Laboratorio de Bioinformática y Expresión Génica, INTA-Universidad de Chile, El Libano 5524, Santiago, Chile.

Laboratorio de Bioinformática y Expresión Génica, INTA-Universidad de Chile, El Libano 5524, Santiago, Chile; Fondap Center for Genome Regulation (CGR), Chile.

出版信息

Gene. 2016 Oct 10;591(1):191-200. doi: 10.1016/j.gene.2016.07.015. Epub 2016 Jul 7.

Abstract

In the early Drosophila melanogaster embryo, the gene regulatory network controlled by Dpp signaling is involved in the subdivision of dorsal ectoderm into the presumptive dorsal epidermis and amnioserosa. In this work, we aimed to identify new Dpp downstream targets involved in dorsal ectoderm patterning. We used oligonucleotide D. melanogaster microarrays to identify the set of genes that are differential expressed between wild type embryos and embryos that overexpress Dpp (nos-Gal4>UAS-dpp) during early stages of embryo development. By using this approach, we identified 358 genes whose relative abundance significantly increased in response to Dpp overexpression. Among them, we found the entire set of known Dpp target genes that function in dorsal ectoderm patterning (zen, doc, hnt, pnr, ush, tup, and others) in addition to several up-regulated genes of unknown functions. Spatial expression pattern of up-regulated genes in response to Dpp overexpression as well as their opposing transcriptional responses to Dpp loss- and gain-of-function indicated that they are new candidate target genes of Dpp signaling pathway. We further analyse one of the candidate genes, CG13653, which is expressed at the dorsal-most cells of the embryo during a restricted period of time. CG13653 orthologs were not detected in basal lineages of Dipterans, which unlike D. melanogaster develop two extra-embryonic membranes, amnion and serosa. We characterized the enhancer region of CG13653 and revealed that CG13653 is directly regulated by Dpp signaling pathway.

摘要

在早期黑腹果蝇胚胎中,由Dpp信号控制的基因调控网络参与将背侧外胚层细分为假定的背侧表皮和羊浆膜。在这项研究中,我们旨在鉴定参与背侧外胚层模式形成的新的Dpp下游靶标。我们使用寡核苷酸黑腹果蝇微阵列来鉴定在胚胎发育早期野生型胚胎和过表达Dpp(nos-Gal4>UAS-dpp)的胚胎之间差异表达的基因集。通过这种方法,我们鉴定出358个基因,其相对丰度因Dpp过表达而显著增加。其中,我们发现了在背侧外胚层模式形成中起作用的所有已知Dpp靶基因(zen、doc、hnt、pnr、ush、tup等)以及几个功能未知的上调基因。对Dpp过表达上调基因的空间表达模式及其对Dpp功能丧失和功能获得的相反转录反应表明,它们是Dpp信号通路的新候选靶基因。我们进一步分析了其中一个候选基因CG13653,它在胚胎发育的特定时期在胚胎最背侧的细胞中表达。在双翅目基础谱系中未检测到CG13653的直系同源物,与黑腹果蝇不同,双翅目基础谱系发育出两种额外的胚胎膜,即羊膜和浆膜。我们对CG13653的增强子区域进行了表征,发现CG13653直接受Dpp信号通路调控。

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