Gene expression analysis of miR-210 over-expressing flies at ZT0 and ZT12. Gene expression analysis of miR-210 over-expressing flies at ZT0 and ZT12

Single microRNAs are usually associated with hundreds of putative target genes that can influence multiple phenotypic traits in Drosophila, ranging from development to behaviour. We investigated the function of Drosophila miR-210 in circadian behaviour by misexpressing it within circadian clock cells. Manipulation of miR-210 expression levels in the PDF (pigment dispersing factor) positive neurons affected the phase of locomotor activity, under both light-dark conditions and constant darkness. PER cyclical expression was not affected in clock neurons, however, when miR-210 was up-regulated, a dramatic alteration in the morphology of PDF ventral lateral neuron (LNv) arborisations was observed. The effect of miR-210 in shaping neuronal projections was confirmed in vitro, using a Drosophila neuronal cell line. A transcriptomic analysis revealed that miR-210 overexpression affects the expression of several genes belonging to pathways related to circadian processes, neuronal development, GTPases signal transduction and photoreception. Collectively, these data reveal the role of miR-210 in modulating circadian outputs in flies and guiding/remodelling PDF positive LNv arborisations and indicate that miR-210 may have pleiotropic effects on the clock, light perception and neuronal development. Overall design: Refer to individual Series

单个微小RNA（microRNAs, miRNAs）通常与数百个推定靶基因相关，这些靶基因可影响果蝇（Drosophila）从发育到行为等多种表型性状。本研究通过在果蝇昼夜节律钟细胞中异位表达miR-210，探究了其在果蝇昼夜节律行为中的功能。在表达色素分散因子（pigment dispersing factor, PDF）的神经元中调控miR-210的表达水平，会影响果蝇在光暗循环及持续黑暗条件下的运动活动时相。在钟细胞中，PER的周期性表达未受影响；但当miR-210被上调时，PDF阳性腹外侧神经元（ventral lateral neuron, LNv）的树突分支形态出现了显著改变。通过使用果蝇神经元细胞系进行体外实验，验证了miR-210对神经元投射形态的调控作用。转录组分析结果显示，miR-210过表达会影响多个基因的表达，这些基因参与的通路涵盖昼夜节律过程、神经元发育、GTP酶信号转导以及光感受。综上，本研究数据揭示了miR-210在调控果蝇昼夜节律输出以及塑造/重塑PDF阳性LNv树突分支中的作用，并表明miR-210可能对生物钟、光感知以及神经元发育具有多效性影响。实验整体设计：详见各独立数据集系列。