Department of Translational Medical Science, Institute of Biosciences and Technology, Texas A&M Health Science Center, Texas A&M University, Houston, TX 77030, USA.
Molecular and Cellular Biology Graduate Program, Arizona State University, Tempe, AZ 85281, USA.
G3 (Bethesda). 2022 May 30;12(6). doi: 10.1093/g3journal/jkac091.
Development of the Caenorhabditis elegans vulva is a classic model of organogenesis. This system, which starts with 6 equipotent cells, encompasses diverse types of developmental event, including developmental competence, multiple signaling events to control precise and faithful patterning of three cell fates, execution and proliferation of specific cell lineages, and a series of sophisticated morphogenetic events. Early events have been subjected to extensive mutational and genetic investigations and later events to cell biological analyses. We infer the existence of dramatically changing profiles of gene expression that accompanies the observed changes in development. Yet, except from serendipitous discovery of several transcription factors expressed in dynamic patterns in vulval lineages, our knowledge of the transcriptomic landscape during vulval development is minimal. This study describes the composition of a vulva-specific transcriptome. We used tissue-specific harvesting of mRNAs via immunoprecipitation of epitope-tagged poly(A) binding protein, PAB-1, heterologously expressed by a promoter known to express GFP in vulval cells throughout their development. The identified transcriptome was small but tightly interconnected. From this data set, we identified several genes with identified functions in development of the vulva and validated more with promoter-GFP reporters of expression. For one target, lag-1, promoter-GFP expression was limited but a fluorescent tag of the endogenous protein revealed extensive expression. Thus, we have identified a transcriptome of C. elegans vulval lineages as a launching pad for exploration of functions of these genes in organogenesis.
秀丽隐杆线虫的生殖道发育是器官发生的经典模型。这个系统从 6 个等能细胞开始,包含多种类型的发育事件,包括发育能力、控制三个细胞命运精确和忠实模式的多种信号事件、特定细胞谱系的执行和增殖,以及一系列复杂的形态发生事件。早期事件已经受到广泛的突变和遗传研究,而后期事件则受到细胞生物学分析的影响。我们推断,伴随着发育过程中观察到的变化,存在着基因表达显著变化的情况。然而,除了在生殖道谱系中以动态模式表达的几个转录因子的偶然发现外,我们对生殖道发育过程中转录组景观的了解还很有限。本研究描述了一个生殖道特异性转录组的组成。我们使用通过免疫沉淀表达 GFP 的 PAB-1 进行组织特异性 mRNA 收获,PAB-1 是通过已知在生殖道细胞整个发育过程中表达 GFP 的启动子异源表达的。鉴定的转录组很小,但紧密互联。从这个数据集,我们鉴定了几个在生殖道发育中具有已知功能的基因,并通过表达的启动子-GFP 报告器进一步验证了更多基因。对于一个靶标 lag-1,启动子-GFP 表达是有限的,但内源性蛋白的荧光标记显示出广泛的表达。因此,我们已经确定了秀丽隐杆线虫生殖道谱系的转录组,作为探索这些基因在器官发生中的功能的起点。