Abstract
MicroRNAs (miRNAs) are key regulators of developmental processes, such as cell fate determination and differentiation. Previous studies showed Dicer knockdown in honeybee embryos disrupt the processing of functional mature miRNAs and impairs embryo patterning. Here we investigated the expression profiles of miRNAs in honeybee embryogenesis and the role of the highly conserved miR-34-5p in the regulation of genes involved in insect segmentation. A total of 221 miRNAs were expressed in honey bee embryogenesis among which 97 mature miRNA sequences have not been observed before. Interestingly, we observed a switch in dominance between the 5-prime and 3-prime arm of some miRNAs in different embryonic stages; however, most miRNAs present one dominant arm across all stages of embryogenesis. Our genome-wide analysis of putative miRNA-target networks and functional pathways indicates miR-34-5p is one of the most conserved and connected miRNAs associated with the regulation of genes involved in embryonic patterning and development. In addition, we experimentally validated that miR-34-5p directly interacts to regulatory elements in the 3′-untranslated regions of pair-rule (even-skipped, hairy, fushi-tarazu transcription factor 1) and cytoskeleton (actin5C) genes. Our study suggests that miR-34-5p may regulate the expression of pair-rule and cytoskeleton genes during early development and control insect segmentation.
Highlights
Segmentation process[29,30]
Transcriptional profile revealed novel miRNAs and arm switching in honey bee embryogenesis
We used high-throughput sequencing of small RNAs to identify miRNAs expressed throughout honey bee embryonic development
Summary
Segmentation process[29,30]. The regulatory role of miRNAs in the segmentation process in the developing embryo is still poorly understood. The highly conserved miR-34-5p is expressed throughout the embryonic development and regulates genes involved in segmentation and morphogenesis. Our results indicate a possible role of miR-34-5p in the regulation of early embryogenesis by directly targeting the pair-rule genes eve, h, ftz-f1, and the structural gene actin5C (act5C). We hypothesize that miR-34-5p regulates the expression of the pair rule genes and act5C in the developing embryo and participates in the determination of cell fate in segmentation process and in the migration of germ-layer forming-cells during embryonic development in insects
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