MiR‐3017b contributes to metamorphosis by targeting sarco/endoplasmic reticulum Ca²⁺ATPase in Tribolium castaneum

Abstract

In recent years, increasing numbers of microRNAs (miRNAs) have been reported to regulate insect metamorphosis. One thousand, one hundred fifty‐four miRNAs have been previously identified from Tribolium castaneum by high‐throughput sequencing; however, little is known about which miRNAs can participate in metamorphosis, leaving the role of miRNAs in regulating the underlying mechanism elusive. Here, we report the participation of miR‐3017b in the metamorphosis of T. castaneum. Temporal profiles revealed that miR‐3017b was highly expressed at the late larval stage, but significantly decreased at the early pupal stage. Overexpression of miR‐3017b caused larval to pupal to adult metamorphosis arrested. Dual‐luciferase reporter assay and miRNA–mRNA interaction assay illustrated that miR‐3017b interacts with the coding sequence of sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) and suppresses its expression. Knockdown of SERCA caused metamorphosis arrested, similar to that observed in miR‐3017b overexpression beetles. Further functional mechanism analyses revealed that 20‐hydroxyecdysone application downregulates miR‐3017b and up‐regulates SERCA expression. The expression level of downstream genes in the 20E pathway was disrupted after overexpressing miR‐3017 and the knockdown of SERCA. These results provided evidence miR‐3017b‐SERCA contributes to metamorphosis by regulating the 20E pathway in T. castaneum. It could advance our understanding of the coordination of 20E and miRNA regulation in insect metamorphosis.

MicroRNA miR‐285 modulates the metamorphosis in Galeruca daurica by targeting Br‐C

“…Moreover, inhibiting miR‐34 resulted in the up‐regulation of the main 20E signaling pathway genes, such as E75 , USP , EcR , Br‐C , and FTZ 25 . Overexpression of miR‐3017b or knockdown of SERCA altered the expression levels of 20E signaling genes such as ECR , USP , HR3 , and Br‐C 29 . These results suggest that 20E controlling the expression of miRNAs might play an important role in promoting the precision of the gene regulation network.…”

“…MiR‐276a‐3p was up‐regulated during the larval–pupal transition and its overexpression reduced developmental growth 27,28 . In T. castaneum , miR‐3017b and its target gene SERCA mediated the metamorphosis by regulating the 20E signaling pathway 29 . In Spodoptera exigua , miR‐4924 regulated larval development and molting 30 .…”

Identification and Characterization of Development-Related microRNAs in the Red Flour Beetle, Tribolium castaneum

“…The GO enrichment analysis showed that these DEmiRNAs mainly participate in the cytoplasm (32), plasma membrane (20), signal transduction (15), and sequence-specific DNA binding transcription factor activity (14) ( Figure 7 B). In the cluster, miR-9a targeting senseless ensures the precise specification of sensory organ precursors in Drosophila [ 55 ], and it also target the Drosophila transcriptional regulator LIM-only to prevent apoptosis during wing development [ 56 ]; miR-8 null flies are smaller in size and defective in insulin signaling in fat body [ 57 ], gain and loss of miR-8 provoked developmental defects reminiscent of impaired Notch signaling [ 58 ], and it also acts as a negative regulator of Wnt signaling to affect eye and wing development in D. melanogaster [ 59 ]; miR-252 has vital roles in growth, development and cell cycle of insect [ 9 , 60 , 61 ]; miR-3017 contributes to larval to pupal to adult metamorphosis by targeting sarco/endoplasmic reticulum Ca 2+ ATPase in T. castaneum [ 62 ]. Mutation of the Drosophila miR-6 cluster, which contains three miR-6 genes, showed a modest reduction in viability to adulthood [ 63 ].…”

Genome‐wide analysis of Panonychus citri microRNAs with a focus on potential insecticidal activity of 4 microRNAs to eggs and nymphs

“…In locusts, agomir-276 and antagomir-276 injection resulted in the advance and the delay of the hatching time, respectively (He et al, 2016). In Tribolium castaneum, overexpression of miR-3017b caused arrest of metamorphosis (Tang et al, 2022). In Drosophila melanogaster, overexpressing miR-315 caused pupation defects and reduced hatching rates (Yuan et al, 2020).…”

“…In Tribolium castaneum , overexpression of miR‐3017b caused arrest of metamorphosis (Tang et al. , 2022). In Drosophila melanogaster , overexpressing miR‐315 caused pupation defects and reduced hatching rates (Yuan et al.…”