The flavonoid 4,4′-dimethoxychalcone promotes autophagy-dependent longevity across species

Ageing constitutes the most important risk factor for all major chronic ailments, including malignant, cardiovascular and neurodegenerative diseases. However, behavioural and pharmacological interventions with feasible potential to promote health upon ageing remain rare. Here we report the identification of the flavonoid 4,4′- dimethoxychalcone (DMC) as a natural compound with anti-ageing properties. External DMC administration extends the lifespan of yeast, worms and flies, decelerates senescence of human cell cultures, and protects mice from prolonged myocardial ischaemia. Concomitantly, DMC induces autophagy, which is essential for its cytoprotective effects from yeast to mice. This pro-autophagic response induces a conserved systemic change in metabolism, operates independently of TORC1 signalling and depends on specific GATA transcription factors. Notably, we identify DMC in the plant Angelica keiskei koidzumi, to which longevity- and health-promoting effects are ascribed in Asian traditional medicine. In summary, we have identified and mechanistically characterised the conserved longevity-promoting effects of a natural anti-ageing drug

A brain-wide form of presynaptic active zone plasticity orchestrates resilience to brain aging in Drosophila

The brain as a central regulator of stress integration determines what is threatening, stores memories, and regulates physiological adaptations across the aging trajectory. While sleep homeostasis seems to be linked to brain resilience, how age-associated changes intersect to adapt brain resilience to life history remains enigmatic. We here provide evidence that a brain-wide form of presynaptic active zone plasticity (“PreScale”), characterized by increases of active zone scaffold proteins and synaptic vesicle release factors, integrates resilience by coupling sleep, longevity, and memory during early aging of Drosophila. PreScale increased over the brain until mid-age, to then decreased again, and promoted the age-typical adaption of sleep patterns as well as extended longevity, while at the same time it reduced the ability of forming new memories. Genetic induction of PreScale also mimicked early aging-associated adaption of sleep patterns and the neuronal activity/excitability of sleep control neurons. Spermidine supplementation, previously shown to suppress early aging-associated PreScale, also attenuated the age-typical sleep pattern changes. Pharmacological induction of sleep for 2 days in mid-age flies also reset PreScale, restored memory formation, and rejuvenated sleep patterns. Our data suggest that early along the aging trajectory, PreScale acts as an acute, brain-wide form of presynaptic plasticity to steer trade-offs between longevity, sleep, and memory formation in a still plastic phase of early brain aging. This study shows that a brain-wide form of presynaptic plasticity ("PreScale") steers trade-offs between longevity, sleep and memory formation in a still plastic phase of early brain aging, illustrating how life strategy manifests at both circuit and synapse levels

Original uncropped immunoblots for Figs 1 and S1.

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PreScale promotes survival over memory formation during early aging.

(A) Direct curve-fitting comparisons between PreScale during early aging from 1d to 30d (Fig 1) and genetic brp titration from 1xBRP to 4xBRP at age 5d [33]. (B and C) Lifespan analysis of flies with BRP titration from 1xBRP to 4xBRP. For male flies (B), n = 394 for 1xBRP (p n = 396 for 3xBRP (p n = 395 for 4xBRP (p wt (n = 393). For female flies (C), n = 400 for 1xBRP (p = 0.017), n = 399 for 3xBRP (ns) and n = 397 for 4xBRP (p wt (n = 399). (D and E) An independent experiment of the lifespan of 2xBRP and 3xBRP flies. For male flies (D), n = 94 for 3xBRP compared to 2xBRP wt (n = 36, p E), n = 98 for 3xBRP compared to 2xBRP wt (n = 59, p F and G) Lifespan analysis of flies with 3xBRP in wake mutant background. For male flies (F), n = 234 for 2xBRP;wake compared to 2xBRP wt control (n = 233, p wake (n = 155, p G), n = 235 for 2xBRP;wake compared to 2xBRP wt control (n = 230, p wake (n = 231, ns). (H and I) Lifespan analysis of flies with 3xBRP in inc mutant background. For male flies (H), n = 236 for inc;2xBRP compared to 2xBRP wt (n = 232, p = 0.009) or compared to inc;3xBRP (n = 212, p I), n = 235 for inc;2xBRP compared to 2xBRP wt (n = 235, p wt is also shown in Fig 1A) or compared to inc;3xBRP (n = 182, ns). (J and K) Lifespan analysis of flies with 3xBRP in atg7 mutant background. For male flies (J), n = 108 for atg7,2xBRP compared to 2xBRP wt (n = 195, p atg7;3xBRP (n = 87, ns). For female flies (K), n = 109 for atg7,2xBRP compared to 2xBRP wt (n = 208, p atg7;3xBRP (n = 87, p L and M) Lifespan analysis of flies with 3xBRP in Alzheimer’s disease model flies (elav>Aβ). For male flies (L), n = 203 for 2xBRP;Aβ compared to elav>mCD8-GFP control (n = 238, p Aβ (n = 151, p = 0.006). For female flies (M), n = 249 for 2xBRP;Aβ compared to elav>mCD8-GFP control (n = 228, p Aβ (n = 241, p = 0.002). Gehan–Breslow–Wilcoxon test with Bonferroni correction for multiple comparisons is shown for all longevity experiments. (N and O) STM for 5d (N) and 30d (O) 2xBRP and 3xBRP flies. n = 19 for 5d 2xBRP, n = 12 for 5d 3xBRP. n = 7–8 for 30d for both groups at 30d. (P and Q) MTM tested 3 h after training for 5d (P) and 30d (Q) 2xBRP and 3xBRP flies. n = 21 for 5d 2xBRP, n = 16 for 5d 3xBRP. n = 8–9 for 30d for both groups at 30d. Student t test is shown. *p S3 Data. BRP, Bruchpilot; MTM, middle-term memory; STM, short-term memory; wt, wild type.</p

Acute deep sleep induced by Gaboxadol/THIP feeding.

(A) Protocol for sleep test of wt female flies treated with different concentrations of THIP at either age 3d or 30d. (B-F) Sleep structure of 5d wt female flies fed with 0.05 mg ml−1 and 0.1 mg ml−1 THIP from measurements over 2–3 days, including sleep profile plotted in 30-min bins (B), daily sleep amount (C), number and duration of sleep episodes (D and E), and sleep latencies (F). n = 64 for untreated control wt flies, n = 32 for both 0.05 mg ml−1 and 0.1 mg ml−1 THIP-treated groups. One-way ANOVA with Bonferroni multiple comparisons test is shown. (G-K) Sleep structure of 30d wt female flies fed with 0.05 mg ml−1 THIP from measurements over 2–3 days, including sleep profile plotted in 30-min bins (G), daily sleep amount (H), number and duration of sleep episodes (I and J), and sleep latencies (K). n = 31 for both groups. Student t test is shown. *p p p S1 Data Sheet. (TIF)</p

Spd supplementation does not affect sleep rebound of 5d young animals.

(A and B) Rationale (A) and protocol (B) for the consequence of Spd supplementation in age-associated alterations of sleep pattern. Early aging trigger PreScale, memory decline, and sleep pattern changes, which might functionally intersect for survival. Spd supplementation was shown to suppress PreScale and memory decline, but its effect on early aging-associated sleep pattern changes was unclear. (C) Sleep profile for 5d wt female flies treated with 5 mM Spd compared to untreated for 3 consecutive days. (D) Normalized cumulative sleep loss during 12-h nighttime sleep deprivation and 24-h sleep rebound. Two-way repeated-measures ANOVA with Fisher LSD test did not detect any significant treatment × time interaction (F(47, 2784) = 0.0038; p > 0.9999) during sleep rebound. (E) Sleep recovered at three different time points after sleep deprivation for 5d 5 mM Spd-treated compared to untreated female flies. n = 29–31 for both groups. Two-way ANOVA with Sidak multiple comparisons is shown. ns, not significant. Error bars: mean ± SEM. Underlying data can be found in S1 Data Sheet. LSD, least significant difference; Spd, spermidine; wt, wild type. (TIF)</p

A model for presynaptic active zone plasticity (PreScale) in executing trade-offs between memory and longevity.

A model for the core AZ scaffold protein BRP-driven PreScale in executing trade-offs between sleep homeostasis, memory formation, and longevity. Early aging provokes PreScale-type plasticity and adaptive sleep pattern changes and subsequently steers trade-offs between memory formation and longevity, which can be mimicked by titrating the gene copies of BRP. Rejuvenation paradigms like spermidine supplementation and Gaboxadol/THIP treatment during early aging eliminate the need of PreScale for regulating adaptive sleep patterns to steer trade-offs between memory and longevity. As a consequence, these paradigms suppress PreScale to allow for new memory formation and lifespan extension. Thus, PreScale likely executes behavioral adaptations and trade-offs during a still plastic phase of early brain aging, illustrating how life strategy manifests on a circuit and synaptic plasticity level. AZ, active zone; BRP, Bruchpilot; Spd, spermidine.</p

Data underlying Fig 5.

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Excessive deep sleep reverses age-associated active zone plasticity/PreScale and boosts lifespan.

(A and B) Rationale (A) and protocol (B) for immunochemistry examinations of fly brains of 0.1 mg ml−1 THIP-treated wt female flies at ages 5d and 30d. Spd supplementation suppresses early aging-associated PreScale, memory decline, and sleep pattern changes, which might functionally intersect for survival. However, though THIP treatment suppresses age-associated memory decline, its effects on PreScale and early aging-associated sleep pattern changes were unclear. (C and D) Confocal images (C) and whole-mount brain staining analysis (D) of BRP in wt female flies at ages 5d and 30d with or without 0.1 mg ml−1 THIP treatment. n = 24–25 for all groups. Scale bar: 50 μm. (E) Protocol for sleep test of wt female flies, which have been treated with 0.1 mg ml−1 THIP for 2 days at age 30d. (F and G) Locomotor walking activity pattern (F) and statistic (G) of 30d wt flies after 2 days of 0.1 mg ml−1 THIP treatment. (H-L) Sleep structure of 30d wt female flies after 2 days of 0.1 mg ml−1 THIP treatment averaged from measurements over 2 days, including sleep profile plotted in 30-min bins (H), daytime and nighttime sleep amount (I), number and duration of sleep episodes (J and K), and sleep latencies (L). n = 90–95 for all groups. (M-P) Protocol (M and O) and 1h MTM (N and P) for 2xBRP and 3xBRP flies, which have been treated with 0.1 mg ml−1 THIP for 2 days at age 2d or 30d. n = 13–16 for all groups. Student t test is shown for comparison between two groups. One-way ANOVA with Bonferroni multiple comparisons test is shown for comparisons of three groups. ** p p Q) Protocol for longevity of 0.05 mg ml−1 THIP-treated wt flies. (R and S) Lifespan analysis of wt male and female flies treated with 0.05 mg ml−1 THIP. For male flies (R), n = 176 for treated compared to untreated (n = 150, ns). For female flies (S), n = 199 for treated compared to untreated (n = 192, p p S7 Data. BRP, Bruchpilot; MTM, middle-term memory; Spd, spermidine; THIP, 4,5,6,7-Tetrahydroisoxazolo[5,4-c]pyridin-3-ol; wt, wild type.</p

Early aging-associated sleep pattern changes of 3xBRP animals.

(A-D) Sleep structure of 1xBRP female flies at age 20d rescued by a transgenic brp copy (gBRP) and averaged from measurements over 2–3 days, including daily sleep amount (A), number and duration of sleep episodes (B and C), and sleep latencies (D). n = 63–64 for all groups. One-way ANOVA with Bonferroni multiple comparisons test is shown. (E and F) Locomotor walking activity distribution across the day (E) and averaged daily total walking activity (F) of 3xBRP compared to 2xBRP female flies at ages 5d, 20d, 30d, and 40d. (G-O) Sleep structure of 3xBRP female flies at ages 5d, 20d, 30d, and 40d averaged from measurements over 2–3 days, including sleep profile plotted in 30-min bins (G), daytime and nighttime sleep amount (H and L), number and duration of sleep episodes (I, J, M, and N), and sleep latencies (K and O). n = 246–247 for 5d, n = 61–63 for 20d, n = 59–62 for 30d, and n = 32 for 40d. Two-way ANOVA with Sidak multiple comparisons is shown. *p p p S1 Data Sheet. (TIF)</p