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The numbers on each image indicate how many worms of the examined ones displayed the indicated phenotype. (Right panels) The gonad of the same animals in the Left panels to indicate the similar developmental stage. (A) Differential interference contrast (DIC) images showing L4 worms recovered from 4-d–starved L1 worms.
S1A indicated a dominant role of intestinal miRNAs in regulating L1 starvation survival. We used a dual-color 3′UTR reporter system (18) to test the computational, prediction-based hypothesis that the 3′UTRs of age-1 and unc-31 are directly regulated by miR-71 (Fig. 3B and Materials and Methods). Among these potential miRNA targets, the predicted miR-71–targeting sites in the 3′UTRs of age-1 and unc-31 are conserved between C.
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On the other hand, the role of a particular miRNA (miR-71) is executed by repressing the expression of many genes in multiple pathways. On one hand, we showed that deletions of a good number of miRNAs have varying impacts on the L1 diapause survival rate, although they may effect the rate through different mechanisms. Instead, many specific physiological functions, such as the starvation-induced stress response, are regulated by a miRNA-target network, often involving multiple miRNAs and a large number of their targets. We found that the known developmental timing genes, hbl-1, lin-42, and lit-1, were at the top of the list (TargetScan). To understand how miR-71 affects VPC division, we searched its predicted targets for potential genes involved in regulating developmental timing. These results indicate that miR-71 plays a significant role in larval development of animals recovering from L1 diapause and likely does so by regulating the expression of components of the insulin receptor/DAF-16 pathway, as well as factors acting downstream, or in parallel to, DAF-16.
Biomass recovery was similar across growth strategies, suggesting that growth-related differences play a minimal role in short-term recovery; however, early regrowth was characterised by contrasting trait shifts. Solidago canadensis exhibited high tolerance to heat and drought, with early biomass and trait recovery, indicating potential for dominance under climate extremes. Biomass fully recovered within one month in both growth strategies, but revery play login leaf traits showed transient shifts, over-recovery in SLA and under-recovery in LDMC, likely reflecting production of new leaf tissues. Please try again in a few minutes.
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That’s why our recovery experts provide a custom treatment plan to fit each individual’s circumstances. You’ve taken the first step on your path to recovery. Images were pseudocolored in Photoshop CS3 (Adobe) and assembled in Illustrator CS3 (Adobe). The primers that were used to amplify the 3′UTR of candidate genes are available upon request. 3′UTRs of genes of interest were cloned into the modified pPD129.57 vector as described previously (18). The data for 3′UTR expression and for VPC timing were analyzed using χ2 test.
- Instead, many specific physiological functions, such as the starvation-induced stress response, are regulated by a miRNA-target network, often involving multiple miRNAs and a large number of their targets.
- MT12993 mir-71(n4115) worms were outcrossed with N2 for four generations before any test except the initial screen.
- The eggs were transferred to plates seeded with HB101 and bleached again 3 d later.
- These results suggest that a significant portion of the miR-71 activities in L1 diapause survival may be devoted to regulating the activities of UNC-31–mediated InsR/PI3K signaling and that the rest of miR-71 activity may regulate UNC-31–independent pathways.
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- Furthermore, a recent study suggests that the expression of certain miRNAs is differentially regulated by starvation-induced dauer diapause (15).
- 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).
miR-71 Is Not Required for Arresting Seam Cell or M-Cell Divisions During L1 Diapause.
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- For example, we observed a robust retarded mutant phenotype in the vulval lineage but did not see obvious defects in seam cell differentiation or alae formation.
- (D) Fractions of worms that carry 3′UTR reporter transgene and show no GFP expression GFP(−), weak GFP expression GFP(+/−), and comparable GFP expression to mCherry GFP(+).
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- Our genetic analysis indicated that for both L1 diapause survival and developmental recovery functions, miR-71 regulates expressions of genes in both the insulin receptor-dependent and -independent pathways.
(A) The mir-71(n4115, lf) mutant displayed severe reduction in L1 starvation survival rate, and the reduced survival rate of mir-71(lf) was suppressed by a reduction-of-function allele of age-1(hx546). (C) The reduced L1 starvation survival rate of ain-1(lf) mutants was significantly suppressed by a null allele of unc-31. 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.
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We thus asked whether miR-71 was required for the reinitiation of developmental programs during the recovery phase after L1 starvation. These results suggest that miR-71 regulates the expression of unc-31 and age-1 through their 3′UTRs. Note that there are extra GFP-positive cells (red arrows) in mir-71(lf) mutants.
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