Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas 66045.
Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas 66045
Genetics. 2020 Dec;216(4):1137-1152. doi: 10.1534/genetics.120.303717. Epub 2020 Oct 28.
Imaginal disc morphogenesis during metamorphosis in provides an excellent model to uncover molecular mechanisms by which hormonal signals effect physical changes during development. The () isoform encodes a transcription factor required for disc morphogenesis in response to 20-hydroxyecdysone, yet how it accomplishes this remains largely unknown. Here, we use functional studies of amorphic mutants and a transcriptional target approach to identify processes driven by and its regulatory targets in leg imaginal discs. mutants fail to properly remodel their basal extracellular matrix (ECM) between 4 and 7 hr after puparium formation. Additionally, mutant discs do not undergo the cell shape changes necessary for leg elongation and fail to elongate normally when exposed to the protease trypsin. RNA-sequencing of wild-type and mutant leg discs identified 717 genes differentially regulated by , including a large number of genes involved in glycolysis, and genes that encode proteins that interact with the ECM. RNA interference-based functional studies reveal that several of these genes are required for adult leg formation, particularly those involved in remodeling the ECM. Additionally, expression is abruptly shut down at the onset of metamorphosis, and expressing it beyond this time results in failure of leg development during the late prepupal and pupal stages. Taken together, our results suggest that is required to drive ECM remodeling, change cell shape, and maintain metabolic activity through the midprepupal stage, but must be switched off to allow expression of pupation genes.
在 变态期间的 imaginal disc 形态发生为揭示激素信号在发育过程中影响物理变化的分子机制提供了一个极好的模型。 () 异构体编码了一个转录因子,它是响应 20-羟基蜕皮酮进行 disc 形态发生所必需的,但它如何完成这一过程在很大程度上仍是未知的。在这里,我们使用 amorphic 突变体的功能研究和转录靶标方法来鉴定 和其调节靶标在 leg imaginal discs 中驱动的过程。 突变体在蛹形成后 4 到 7 小时之间无法正确重塑其基底细胞外基质 (ECM)。此外, 突变盘不会发生 leg 伸长所必需的细胞形状变化,并且当暴露于蛋白酶胰蛋白酶时不会正常伸长。野生型和 突变体 leg disc 的 RNA-seq 鉴定了 717 个由 差异调控的基因,包括大量参与糖酵解的基因和与 ECM 相互作用的蛋白质编码基因。基于 RNA 干扰的功能研究表明,这些基因中的几个基因对于成虫腿的形成是必需的,特别是那些涉及重塑 ECM 的基因。此外, 在变态开始时突然关闭表达,并且在此时间之后表达它会导致在晚期 prepupal 和蛹期期间腿发育失败。总之,我们的结果表明, 在 midprepupal 阶段需要驱动 ECM 重塑、改变细胞形状和维持代谢活性,但必须关闭以允许蛹化基因的表达。