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Deletion of microRNA-80 activates dietary restriction to extend C. elegans healthspan and lifespan.

Vora M, Shah M, Ostafi S, Onken B, Xue J, Ni JZ, Gu S, Driscoll M - PLoS Genet. (2013)

Bottom Line: Caloric/dietary restriction (CR/DR) can promote longevity and protect against age-associated disease across species.The molecular mechanisms coordinating food intake with health-promoting metabolism are thus of significant medical interest.Under food limitation, lowered miR-80 levels directly or indirectly increase CBP-1 protein levels to engage metabolic loops that promote DR.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology and Biochemistry, Nelson Biological Laboratories, Rutgers, The State University of New Jersey, Piscataway, New Jersey, United States of America.

ABSTRACT
Caloric/dietary restriction (CR/DR) can promote longevity and protect against age-associated disease across species. The molecular mechanisms coordinating food intake with health-promoting metabolism are thus of significant medical interest. We report that conserved Caenorhabditis elegans microRNA-80 (mir-80) is a major regulator of the DR state. mir-80 deletion confers system-wide healthy aging, including maintained cardiac-like and skeletal muscle-like function at advanced age, reduced accumulation of lipofuscin, and extended lifespan, coincident with induction of physiological features of DR. mir-80 expression is generally high under ad lib feeding and low under food limitation, with most striking food-sensitive expression changes in posterior intestine. The acetyltransferase transcription co-factor cbp-1 and interacting transcription factors daf-16/FOXO and heat shock factor-1 hsf-1 are essential for mir-80(Δ) benefits. Candidate miR-80 target sequences within the cbp-1 transcript may confer food-dependent regulation. Under food limitation, lowered miR-80 levels directly or indirectly increase CBP-1 protein levels to engage metabolic loops that promote DR.

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Related in: MedlinePlus

hsf-1 is needed for the fluorimetric DR signature and longevity phenotypes of mir-80(Δ).Fig. 5A. hsf-1(RNAi) in the mir-80(Δ) background reverses the DR Exmax shift. We grew age-synchronized animals under standard RNAi feeding conditions (20°C, HT115) and measured age pigments at Day 4 (50 animals per RNAi clone). We recorded Exmax as the highest peak detected by the Datamax software package suite (Horiba Scientific). Black bar, WT+ empty vector RNAi; red bar, mir-80(Δ)+empty vector RNAI; grey bar, mir-80(Δ)+hsf-1(RNAi). Graphs represent cumulative data from 3 independent trials. Error bars represent ±S.E.M. Data were compared using 2-tailed Student's T-test (** p<0.001). Note that hsf-1(RNAi) treatment of WT does not change Exmax (data not shown). Fig. 5B. hsf-1(RNAi) in the mir-80(Δ) background partially counters the low age pigment level phenotype of mir-80(Δ). We grew age-synchronized animals under standard conditions (20°C, HT115) and measured total age pigment fluorescence, normalized to total tryptophan fluorescence as in [19] (Day 4 post-hatching, 50 animals per RNAi clone). Black bar, WT+ empty vector RNAi; red bar, mir-80(Δ)+empty vector RNAi; grey bar, mir-80(Δ)+hsf-1(RNAi). Graphs represent cumulative data from 3 independent trials. Error bars represent ±S.E.M. Data were compared using 2-tailed Student's T-test (*** p<0.0001, * p<0.05 compared to mir-80(Δ) empty vector). Note that hsf-1(RNAi) treatment of WT does not change age pigment scores at day 4 (data not shown). Fig. 5C. hsf-1 is required for mir-80(Δ)-induced longevity. We grew age-synchronized animals under standard conditions with low levels of FUDR to prevent progeny production (20°C, OP50-1, 50 uM FuDR). At day 9, we placed 10 healthy animals per plate, ≥40 per strain per trial, and we scored viability as movement away from pick touch at the indicated days. The graphs represent data combined from 3 independent trials. Statistics are calculated using the Log-rank Test. Error bars indicate ± S.E.M. The mir-80(Δ); hsf-1(sy441) double mutant is shorter lived than mir-80(Δ) (p<0.0001). Because RNAi knockdown is inefficient the nervous system (see [59]), the profound effects of hsf-1(RNAi) suggest that critical hsf-1 and mir-80 regulation occurs outside of the C. elegans nervous system.
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pgen-1003737-g005: hsf-1 is needed for the fluorimetric DR signature and longevity phenotypes of mir-80(Δ).Fig. 5A. hsf-1(RNAi) in the mir-80(Δ) background reverses the DR Exmax shift. We grew age-synchronized animals under standard RNAi feeding conditions (20°C, HT115) and measured age pigments at Day 4 (50 animals per RNAi clone). We recorded Exmax as the highest peak detected by the Datamax software package suite (Horiba Scientific). Black bar, WT+ empty vector RNAi; red bar, mir-80(Δ)+empty vector RNAI; grey bar, mir-80(Δ)+hsf-1(RNAi). Graphs represent cumulative data from 3 independent trials. Error bars represent ±S.E.M. Data were compared using 2-tailed Student's T-test (** p<0.001). Note that hsf-1(RNAi) treatment of WT does not change Exmax (data not shown). Fig. 5B. hsf-1(RNAi) in the mir-80(Δ) background partially counters the low age pigment level phenotype of mir-80(Δ). We grew age-synchronized animals under standard conditions (20°C, HT115) and measured total age pigment fluorescence, normalized to total tryptophan fluorescence as in [19] (Day 4 post-hatching, 50 animals per RNAi clone). Black bar, WT+ empty vector RNAi; red bar, mir-80(Δ)+empty vector RNAi; grey bar, mir-80(Δ)+hsf-1(RNAi). Graphs represent cumulative data from 3 independent trials. Error bars represent ±S.E.M. Data were compared using 2-tailed Student's T-test (*** p<0.0001, * p<0.05 compared to mir-80(Δ) empty vector). Note that hsf-1(RNAi) treatment of WT does not change age pigment scores at day 4 (data not shown). Fig. 5C. hsf-1 is required for mir-80(Δ)-induced longevity. We grew age-synchronized animals under standard conditions with low levels of FUDR to prevent progeny production (20°C, OP50-1, 50 uM FuDR). At day 9, we placed 10 healthy animals per plate, ≥40 per strain per trial, and we scored viability as movement away from pick touch at the indicated days. The graphs represent data combined from 3 independent trials. Statistics are calculated using the Log-rank Test. Error bars indicate ± S.E.M. The mir-80(Δ); hsf-1(sy441) double mutant is shorter lived than mir-80(Δ) (p<0.0001). Because RNAi knockdown is inefficient the nervous system (see [59]), the profound effects of hsf-1(RNAi) suggest that critical hsf-1 and mir-80 regulation occurs outside of the C. elegans nervous system.

Mentions: To identify genes required for mir-80(Δ)-regulated DR, we used RNAi to knockdown genes previously implicated in DR lifespan benefits, hypothesizing that genes required for mir-80(Δ) DR should be needed for the Exmax shift and low age pigment levels typical of multiple DR states. Of the 18 genes we screened, we found that RNAi knockdown of transcription factors daf-16/FOXO, heat shock transcription factor hsf-1, and CREB binding protein homolog cbp-1 modulated both the Exmax shift and low age pigment levels of mir-80(Δ) (Tables S1, S2, Figs. 4A,B; 5A,B; 6A,B).


Deletion of microRNA-80 activates dietary restriction to extend C. elegans healthspan and lifespan.

Vora M, Shah M, Ostafi S, Onken B, Xue J, Ni JZ, Gu S, Driscoll M - PLoS Genet. (2013)

hsf-1 is needed for the fluorimetric DR signature and longevity phenotypes of mir-80(Δ).Fig. 5A. hsf-1(RNAi) in the mir-80(Δ) background reverses the DR Exmax shift. We grew age-synchronized animals under standard RNAi feeding conditions (20°C, HT115) and measured age pigments at Day 4 (50 animals per RNAi clone). We recorded Exmax as the highest peak detected by the Datamax software package suite (Horiba Scientific). Black bar, WT+ empty vector RNAi; red bar, mir-80(Δ)+empty vector RNAI; grey bar, mir-80(Δ)+hsf-1(RNAi). Graphs represent cumulative data from 3 independent trials. Error bars represent ±S.E.M. Data were compared using 2-tailed Student's T-test (** p<0.001). Note that hsf-1(RNAi) treatment of WT does not change Exmax (data not shown). Fig. 5B. hsf-1(RNAi) in the mir-80(Δ) background partially counters the low age pigment level phenotype of mir-80(Δ). We grew age-synchronized animals under standard conditions (20°C, HT115) and measured total age pigment fluorescence, normalized to total tryptophan fluorescence as in [19] (Day 4 post-hatching, 50 animals per RNAi clone). Black bar, WT+ empty vector RNAi; red bar, mir-80(Δ)+empty vector RNAi; grey bar, mir-80(Δ)+hsf-1(RNAi). Graphs represent cumulative data from 3 independent trials. Error bars represent ±S.E.M. Data were compared using 2-tailed Student's T-test (*** p<0.0001, * p<0.05 compared to mir-80(Δ) empty vector). Note that hsf-1(RNAi) treatment of WT does not change age pigment scores at day 4 (data not shown). Fig. 5C. hsf-1 is required for mir-80(Δ)-induced longevity. We grew age-synchronized animals under standard conditions with low levels of FUDR to prevent progeny production (20°C, OP50-1, 50 uM FuDR). At day 9, we placed 10 healthy animals per plate, ≥40 per strain per trial, and we scored viability as movement away from pick touch at the indicated days. The graphs represent data combined from 3 independent trials. Statistics are calculated using the Log-rank Test. Error bars indicate ± S.E.M. The mir-80(Δ); hsf-1(sy441) double mutant is shorter lived than mir-80(Δ) (p<0.0001). Because RNAi knockdown is inefficient the nervous system (see [59]), the profound effects of hsf-1(RNAi) suggest that critical hsf-1 and mir-80 regulation occurs outside of the C. elegans nervous system.
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pgen-1003737-g005: hsf-1 is needed for the fluorimetric DR signature and longevity phenotypes of mir-80(Δ).Fig. 5A. hsf-1(RNAi) in the mir-80(Δ) background reverses the DR Exmax shift. We grew age-synchronized animals under standard RNAi feeding conditions (20°C, HT115) and measured age pigments at Day 4 (50 animals per RNAi clone). We recorded Exmax as the highest peak detected by the Datamax software package suite (Horiba Scientific). Black bar, WT+ empty vector RNAi; red bar, mir-80(Δ)+empty vector RNAI; grey bar, mir-80(Δ)+hsf-1(RNAi). Graphs represent cumulative data from 3 independent trials. Error bars represent ±S.E.M. Data were compared using 2-tailed Student's T-test (** p<0.001). Note that hsf-1(RNAi) treatment of WT does not change Exmax (data not shown). Fig. 5B. hsf-1(RNAi) in the mir-80(Δ) background partially counters the low age pigment level phenotype of mir-80(Δ). We grew age-synchronized animals under standard conditions (20°C, HT115) and measured total age pigment fluorescence, normalized to total tryptophan fluorescence as in [19] (Day 4 post-hatching, 50 animals per RNAi clone). Black bar, WT+ empty vector RNAi; red bar, mir-80(Δ)+empty vector RNAi; grey bar, mir-80(Δ)+hsf-1(RNAi). Graphs represent cumulative data from 3 independent trials. Error bars represent ±S.E.M. Data were compared using 2-tailed Student's T-test (*** p<0.0001, * p<0.05 compared to mir-80(Δ) empty vector). Note that hsf-1(RNAi) treatment of WT does not change age pigment scores at day 4 (data not shown). Fig. 5C. hsf-1 is required for mir-80(Δ)-induced longevity. We grew age-synchronized animals under standard conditions with low levels of FUDR to prevent progeny production (20°C, OP50-1, 50 uM FuDR). At day 9, we placed 10 healthy animals per plate, ≥40 per strain per trial, and we scored viability as movement away from pick touch at the indicated days. The graphs represent data combined from 3 independent trials. Statistics are calculated using the Log-rank Test. Error bars indicate ± S.E.M. The mir-80(Δ); hsf-1(sy441) double mutant is shorter lived than mir-80(Δ) (p<0.0001). Because RNAi knockdown is inefficient the nervous system (see [59]), the profound effects of hsf-1(RNAi) suggest that critical hsf-1 and mir-80 regulation occurs outside of the C. elegans nervous system.
Mentions: To identify genes required for mir-80(Δ)-regulated DR, we used RNAi to knockdown genes previously implicated in DR lifespan benefits, hypothesizing that genes required for mir-80(Δ) DR should be needed for the Exmax shift and low age pigment levels typical of multiple DR states. Of the 18 genes we screened, we found that RNAi knockdown of transcription factors daf-16/FOXO, heat shock transcription factor hsf-1, and CREB binding protein homolog cbp-1 modulated both the Exmax shift and low age pigment levels of mir-80(Δ) (Tables S1, S2, Figs. 4A,B; 5A,B; 6A,B).

Bottom Line: Caloric/dietary restriction (CR/DR) can promote longevity and protect against age-associated disease across species.The molecular mechanisms coordinating food intake with health-promoting metabolism are thus of significant medical interest.Under food limitation, lowered miR-80 levels directly or indirectly increase CBP-1 protein levels to engage metabolic loops that promote DR.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology and Biochemistry, Nelson Biological Laboratories, Rutgers, The State University of New Jersey, Piscataway, New Jersey, United States of America.

ABSTRACT
Caloric/dietary restriction (CR/DR) can promote longevity and protect against age-associated disease across species. The molecular mechanisms coordinating food intake with health-promoting metabolism are thus of significant medical interest. We report that conserved Caenorhabditis elegans microRNA-80 (mir-80) is a major regulator of the DR state. mir-80 deletion confers system-wide healthy aging, including maintained cardiac-like and skeletal muscle-like function at advanced age, reduced accumulation of lipofuscin, and extended lifespan, coincident with induction of physiological features of DR. mir-80 expression is generally high under ad lib feeding and low under food limitation, with most striking food-sensitive expression changes in posterior intestine. The acetyltransferase transcription co-factor cbp-1 and interacting transcription factors daf-16/FOXO and heat shock factor-1 hsf-1 are essential for mir-80(Δ) benefits. Candidate miR-80 target sequences within the cbp-1 transcript may confer food-dependent regulation. Under food limitation, lowered miR-80 levels directly or indirectly increase CBP-1 protein levels to engage metabolic loops that promote DR.

Show MeSH
Related in: MedlinePlus