Institut de Biologia Molecular de Barcelona, CSIC, Barcelona, Spain.
PLoS One. 2011;6(12):e28985. doi: 10.1371/journal.pone.0028985. Epub 2011 Dec 22.
The Drosophila transcription factor Tramtrack (Ttk) is involved in a wide range of developmental decisions, ranging from early embryonic patterning to differentiation processes in organogenesis. Given the wide spectrum of functions and pleiotropic effects that hinder a comprehensive characterisation, many of the tissue specific functions of this transcription factor are only poorly understood. We recently discovered multiple roles of Ttk in the development of the tracheal system on the morphogenetic level. Here, we sought to identify some of the underlying genetic components that are responsible for the tracheal phenotypes of Ttk mutants. We therefore profiled gene expression changes after Ttk loss- and gain-of-function in whole embryos and cell populations enriched for tracheal cells. The analysis of the transcriptomes revealed widespread changes in gene expression. Interestingly, one of the most prominent gene classes that showed significant opposing responses to loss- and gain-of-function was annotated with functions in chitin metabolism, along with additional genes that are linked to cellular responses, which are impaired in ttk mutants. The expression changes of these genes were validated by quantitative real-time PCR and further functional analysis of these candidate genes and other genes also expected to control tracheal tube size revealed at least a partial explanation of Ttk's role in tube size regulation. The computational analysis of our tissue-specific gene expression data highlighted the sensitivity of the approach and revealed an interesting set of novel putatively tracheal genes.
果蝇转录因子 Tramtrack(Ttk)参与了广泛的发育决策,从早期胚胎模式形成到器官发生中的分化过程。鉴于该转录因子具有广泛的功能和多效性影响,阻碍了全面的特征描述,因此,许多组织特异性功能尚未得到充分理解。我们最近在气管系统的形态发生水平上发现了 Ttk 的多种作用。在这里,我们试图确定一些负责 Ttk 突变体气管表型的遗传组成部分。因此,我们在整个胚胎和富含气管细胞的细胞群体中,对 Ttk 功能丧失和功能获得后的基因表达变化进行了分析。转录组的分析显示基因表达广泛变化。有趣的是,在 Ttk 功能丧失和功能获得后,最显著的基因类别之一是与几丁质代谢相关的功能注释,以及与细胞反应相关的其他基因,这些基因在 ttk 突变体中受损。通过定量实时 PCR 验证了这些基因的表达变化,并对这些候选基因和其他也预期控制气管管腔大小的基因进行了进一步的功能分析,至少部分解释了 Ttk 在管腔大小调节中的作用。我们组织特异性基因表达数据的计算分析突出了该方法的敏感性,并揭示了一组有趣的新的可能与气管相关的基因。