Dissecting Metabolic Control of Behaviors and Physiology During Aging in Drosophila

Aging disrupts physiological and behavioral homeostasis, largely driven by one-carbon metabolism, mitochondrial dysfunction, energy sensing, and metabolic imbalance. To elucidate the roles of conserved metabolic, energy sensing, and mitochondrial genes in age-related decline, we employed genetic manipulations in vivo using Drosophila melanogaster models, in a cell-autonomous and non-cell-autonomous manner. By using panneuronal and indirect flight muscle (IFM)- specific drivers, we assessed the impact of gene knockdown or overexpression on sleep-circadian rhythms, locomotion, and lipid metabolism in a cell-autonomous and non-cell-autonomous manner to address bidirectional neuro-muscle communications. Knockdown of genes such as SdhD, Marf, and Gnmt leads to decrease in flight performance especially in 6 weeks with both the drivers. Which demonstrates cell-autonomous and non-cell autonomous effects of these genes. Negative geotaxis with panneuronal knockdown of Adsl, Gnmt, SdhD, Marf genes showed reduced locomotor performance in age-dependent manner consolidating their non-cell autonomous role and neuro-muscular interaction. Whereas mAcon1, LSD2, Ampkα, Ald, Adsl genes showed reduced flight performance with only IFM specific driver emphasizing the cell-autonomous role. Panneuronal knockdown of Ald, GlyP, mAcon1, and Gnmt genes showed increased total sleep, reduced activity, while Adsl and Ogdh knockdown led to sleep fragmentation, in a mid-age suggests cell autonomous impact. Functional analysis of AMPK signaling via overexpression and knockdown of Ampkα, as well as expression of the yeast ortholog SNF1A and its kinase-dead mutant, revealed kinase-dependent, age- and tissue-specific modulation of sleep and activity rhythms. Lipid analysis showed that panneuronal overexpression of Ampkα altered lipid droplet number and size in the brain, indicating disrupted lipid homeostasis during aging. These findings establish Ampkα as a central regulator of behavioral and metabolic aging, linking neuronal energy sensing, motor function, and lipid dynamics, and offer mechanistic insights into tissue-specific metabolic regulation with potential relevance for interventions targeting age-related decline and neurodegeneration.