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深度 mRNA 测序分析以捕获斑马鱼胚胎和幼虫的转录组图谱。

Deep mRNA sequencing analysis to capture the transcriptome landscape of zebrafish embryos and larvae.

机构信息

State Key Laboratory of Genetic Engineering, Institute of Plant Biology, Center of Evolutionary Biology, School of Life Science, Fudan University, Shanghai, China.

出版信息

PLoS One. 2013 May 20;8(5):e64058. doi: 10.1371/journal.pone.0064058. Print 2013.

Abstract

Transcriptome analysis is a powerful tool to obtain large amount genome-scale gene expression profiles. Despite its extensive usage to diverse biological problems in the last decade, transcriptomic researches approaching the zebrafish embryonic development have been very limited. Several recent studies have made great progress in this direction, yet the large gap still exists, especially regarding to the transcriptome dynamics of embryonic stages from early gastrulation onwards. Here, we present a comprehensive analysis about the transcriptomes of 9 different stages covering 7 major periods (cleavage, blastula, gastrula, segmentation, pharyngula, hatching and early larval stage) in zebrafish development, by recruiting the RNA-sequencing technology. We detected the expression for at least 24,065 genes in at least one of the 9 stages. We identified 16,130 genes that were significantly differentially expressed between stages and were subsequently classified into six clusters. Each revealed gene cluster had distinct expression patterns and characteristic functional pathways, providing a framework for the understanding of the developmental transcriptome dynamics. Over 4000 genes were identified as preferentially expressed in one of the stages, which could be of high relevance to stage-specific developmental and molecular events. Among the 68 transcription factor families active during development, most had enhanced average expression levels and thus might be crucial for embryogenesis, whereas the inactivation of the other families was likely required by the activation of the zygotic genome. We discussed our RNA-seq data together with previous findings about the Wnt signaling pathway and some other genes with known functions, to show how our data could be used to advance our understanding about these developmental functional elements. Our study provides ample information for further study about the molecular and cellular mechanisms underlying vertebrate development.

摘要

转录组分析是获取大量基因组规模基因表达谱的强大工具。尽管在过去十年中,它被广泛应用于各种生物学问题,但针对斑马鱼胚胎发育的转录组研究非常有限。最近的几项研究在这方面取得了很大进展,但仍存在很大差距,特别是在早期原肠胚以后的胚胎发育阶段的转录组动态方面。在这里,我们通过 RNA 测序技术,对涵盖斑马鱼发育 7 个主要时期(分裂、囊胚、原肠胚、分节、咽胚、孵化和早期幼体阶段)的 9 个不同阶段的转录组进行了全面分析。我们至少在 9 个阶段中的 1 个阶段中检测到了至少 24065 个基因的表达。我们鉴定了 16130 个在不同阶段之间差异表达的基因,并随后将其分为 6 个簇。每个揭示的基因簇都具有独特的表达模式和特征性功能途径,为理解发育转录组动态提供了框架。4000 多个基因被鉴定为在一个阶段中优先表达,这些基因可能与阶段特异性发育和分子事件高度相关。在发育过程中活跃的 68 个转录因子家族中,大多数的平均表达水平增强,因此可能对胚胎发生至关重要,而其他家族的失活可能是合子基因组激活所必需的。我们将我们的 RNA-seq 数据与以前关于 Wnt 信号通路和一些其他具有已知功能的基因的发现一起讨论,以展示我们的数据如何用于推进我们对这些发育功能元件的理解。我们的研究为进一步研究脊椎动物发育的分子和细胞机制提供了丰富的信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a3d8/3659048/e85e8a9caf02/pone.0064058.g001.jpg

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