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• Images were pseudocolored in Photoshop CS3 (Adobe) and assembled in Illustrator CS3 (Adobe).
• Duo Mobile’s restore functionality lets you back up Duo-protected accounts and third-party OTP accounts (such as Google or Facebook) for recovery to the same device or to a new device.
• The reporter is strongly expressed in H and V cells in both wild-type and mir-71(lf) worms.
• Reduction-of-function mutation (rf) in the age-1/PI3 kinase gene, age-1(hx546), made worms long-lived in the L1 starvation assay and was able to suppress the reduced L1 survival rate of mir-71(lf); the rate of the double mutants was comparable to that of wild type (Fig. 2A).
• Compromising overall miRNA function dramatically reduces the survival rate of L1 worms in starvation-induced diapause, and the effect can be significantly suppressed by an age-1/PI3K mutation.
• Here we show that compromising overall microRNA (miRNA) functions or mutating certain individual miRNAs impairs the long-term survival of nematodes during starvation-induced L1 diapause.
• We found that the 3′UTRs of several genes of the InsR pathway, including unc-31, age-1, pdk-1, akt-2, and sgk-1, contain predicted miR-71 targeting sites (as predicted by TargetScan and mirWIP).

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The presented results indicate that interactions between multiple miRNAs and likely a large number of their mRNA targets in multiple pathways regulate the response to starvation-induced L1 diapause. Numerous animal species across multiple phyla enter developmental arrest for long-term survival in unfavorable environments and resume development upon stress removal. Such lagged trait recovery, combined with rapid invasive recovery, suggests potential for longer-term shifts in grassland composition and function.
MiR-71 regulates vulval cell division during recovery of starved L1 worms. These results indicate that miR-71 is not essential for arresting seam cell or M-cell divisions during L1 diapause, suggesting that miR-71 function is distinct revery play login from DAF-16 function. DAF-16 (the FOXO homolog in C. elegans) has been shown to play an important role in cell cycle arrest and developmental progression partly by promoting cki-1 expression in some somatic cells during L1 arrest (2).

• To compare the survival rates between strains, we simulated the survival rate of each genotype to 100 arbitrary “individual worms” and performed the log-rank test in Graphpad Prism 4.
• These results compelled us to examine specific interactions between individual miRNAs and their targets to gain mechanistic insights.
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• Unlike classical heterochronic miRNAs such as lin-4 and let-7, the role of miR-71 in vulval cell division is essential in animals recovering from starvation-induced L1 diapause, but not in animals hatched on plates with food.
• To determine the functional relationship of miR-71 with LIN-42 and LIT-1, mir-71(lf); lin-42(lf) L1 worms were starved for 4 d and recovered on lit-1(RNAi) plates.
• However, the mechanisms that coordinate the long-term survival, overall developmental arrest, and reinitiation remain to be investigated.
• (H and I) Fluorescence images (H) and statistical data (I) showing that the M cell diveded in fed animals but remained undivided in 4-, 7-, or 11-d–starved L1 wild-type and mir-71(lf) worms.

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To investigate the roles of miRNAs in animal survival during starvation-induced L1 diapause, we impaired the overall miRISC function with loss-of-function (lf) mutants of ain-1 (ku322, ku425, and tm3681) and ain-2(tm2432) and examined their L1 starvation survival rate (Materials and Methods). The strong suppression of the mir-71(lf) defect by hbl-1(RNAi), and the relatively weak effect of miR-71 on hbl-1 expression, are consistent with the idea that miR-71 exerts its role by modulating activities of multiple genes related to hbl-1 function in developmental timing. In contrast, the nuclear-localized GFP expression under the control of the 3′UTR of age-1(Fig. 3 C and D) or unc-31 (Fig. 3 E and F) was strongly repressed in the control worms, but prominently derepressed in mir-71(lf) mutant worms. If the 3′UTR of age-1 or unc-31 is repressed by miR-71, the GFP expression will be repressed in tissues where miR-71 is expressed in wild-type worms, but derepressed in the same tissues of mir-71(lf) worms.