Fig. 4: Activation of neuronal AMPK in early life is required for the MAE to program offspring body size in adulthood.

a Adult body length measurements in offspring born to D2 aak-2(ok524) mutant worms treated with 5 μM rotenone (Rot) or 50 mM metformin (Met) for 12 h. ddH₂O served as the vehicle control (Veh). b Adult body length changes in WT animals born to D2 mothers in response to early-life treatment from the larval 1 (L1) stage to the L4 stage with 1 mM AICAR. c Body length comparison between OD2 and OD5 aak-2(ok524) mutant animals. d Tissue-specific role of AMPK in mediating the MAE on offspring body size. The experiment was performed with ZIF-1-mediated protein degradation of GFP-fused AAK-2. The success of tissue-specific depletion of AMPK was indicated by loss of GFP fluorescence in the indicated tissues (white arrows) in the left images. The adult body lengths of OD2 and OD5 animals with the indicated tissue-specific AMPK depletion are compared as listed in the right panel. The scale bars represent 20 μm. e Model of the mitochondria-AMPK interplay in mediating the shaping of offspring adult size by MAE. Dots in box plots represent worm numbers. Box plots in a–d: the centerline is the median, the box range shows the 25th–75th percentiles, and the whiskers indicate the minimum–maximum values. The numbers (n) of total tested animals for each group are shown beneath the x-axis. The box plots (a–d) were analyzed by unpaired t-test. Biological replicates: 3 (a–d).