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• carlos.solis
• 20 septiembre, 2024
• Reveryplay
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(B) Survival rate of single and double mutants to indicate the functional relationship between ain-1 and age-1. The two ain-1 loss-of-function alleles displayed significant reductions in L1 starvation survival rate. We further found that this survival rate reduction of ain-1 mutants was overcome by ectopic expression of the AIN-2 protein in the intestine but not in the muscle (Fig. 1A and Fig. S1A). We found that ain-1 but not ain-2 mutants displayed a significant reduction in L1 starvation survival rate compared with that of wild type (Fig. 1 A and D).
Furthermore, a recent study suggests that the expression of certain miRNAs is differentially regulated by starvation-induced dauer diapause (15). Consistent with these ideas, several recent lines of evidence suggest that miRNA let-7 and the heterochronic genes lin-42 and hbl-1 are required to regulate the starvation-induced dauer diapause (10–12) and that a number of miRNAs including lin-4 and mir-71 are involved in regulating life span (13, 14). Furthermore, worms that are long-lived due to dietary restriction or decreased mitochondrial respiratory rates are short-lived during L1 diapause, suggesting that the mechanisms controlling L1 starvation survival are different at least in some aspects from those controlling aging (3).
Duo Mobile cannot recover access to those accounts without a backup. Be sure to enable third-party account backup and restore if you use Duo Mobile to generate passcodes for logging into applications like Instagram, Facebook, Snapchat, or other web services. 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. This result suggests that the high expression of miR-71 during L1 diapause is induced or maintained by other signaling pathways. We asked whether the expression of miR-71 was regulated by DAF-16, which is required during L1 diapause for long-term survival (2).
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.

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• (E) Fluorescence and DIC images showing that the unc-31 3′UTR reporter was repressed in mir-71(+)worms (2/2 transgenic lines) but not in mir-71(lf) worms (4/4 transgenic lines).
• (F) Fluorescence and DIC images showing that an hbl-1 3′UTR reporter was repressed in mir-71(+) worms and slightly derepressed in mir-71(lf) mutants.
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• The coordinated entrance into developmental arrest, long-term survival, and the reinitiation of development upon food availability are important biological processes to investigate.
• Solidago canadensis exhibited high tolerance to heat and drought, with early biomass and trait recovery, indicating potential for dominance under climate extremes.
• Our data provide the experimental evidence that two components of the InsR pathway are likely direct targets of miR-71 in its role in a specific physiological process, L1 diapause (see a model in Fig. S5).

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Moreover, the expression of hbl-1 is repressed by let-7 family miRNAs at L3 during normal development, and the hyperactivity of hbl-1 caused by failure of miRNA regulation leads to retarded development (26). The reporter construct, the control plasmid, and a transformation marker plasmid were coinjected into worms to generate the extrachromosomal arrays for analysis. Elegans and Caenorhabditis briggsae, leading us to focus further analyses on these two genes. We further examined the functional relationship between miR-71 and DAF-16, a FOXO transcription factor acting critically and negatively downstream of AGE-1/PI3K in the InsR pathway.

Intestinal miRNAs Play Critical Roles in L1 Starvation Survival.

• The eggs were transferred to plates seeded with HB101 and bleached again 3 d later.
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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.
• Furthermore, a recent study suggests that the expression of certain miRNAs is differentially regulated by starvation-induced dauer diapause (15).
• 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.
• 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.

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L1 starvation assay was adapted from a previously described protocol (3). Worms strains were grown and maintained at 20 °C as described (29). This result is consistent with the observation that miR-71 is specifically required for the starvation-induced stress response (Fig. S5). 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. It seems plausible that miRNAs that control developmental timing are also involved in regulating the metabolic rate through repressing the InsR pathway activity.
Mutating miR-71 drastically reduces the survival rate of animals in L1 diapause, and the effect can be suppressed by mutations of insulin receptor pathway genes age-1 and unc-31. Among short-lived miRNA mutants, a mir-71 deletion mutant, mir-71(n4115) (referred to as mir-71(lf) hereafter), displayed a severe reduction in L1 starvation survival rate (Table S1 and Fig. 2A). We found that the reduced survival rate of ain-1 was suppressed by either reduction of age-1 function or loss of unc-31 function (Fig. 1 B and C), suggesting that a significant portion of the overall miRNA functions in L1 diapause is upstream of, or in parallel to, the InsR pathway. In this study, we addressed the questions of whether and how miRNAs impact developmental arrest and the long-term survival of early L1 stage worms in response to food starvation. 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. Third-party accounts will also be restored if third-party backup was enabled on the old device.