Metformin has no significant effect on the growth, development, or activity of zebrafish larvae. (a) The chemical formula of metformin. (b) Survival rate curve of zebrafish larvae from 0 to 96 hpf (%survival, n = 15). (c) Tail swing frequency of 30 hpf zebrafish larvae (swing per minute, n = 15). (d) Heartbeat of 24 hpf zebrafish larvae (beats per minute, n = 30). (e) Heartbeat of 48 hpf zebrafish larvae (beats per minute, n = 30). (f) Heartbeat of 24 hpf zebrafish larvae (beats per minute, n = 30). (g) Statistical graph of the fork length of 96 hpf zebrafish larvae (mm, n = 15). (h) Morphological images of 96 hpf zebrafish larvae observed under a stereomicroscope (scale bar: 350 μm). There were no significant differences in morphology between the 50 μM metformin-treated group and the control group. The data were analyzed using one-way ANOVA and the log-rank test. All the experiments were repeated three times. (i,j) The behavioral rhythms of the control group (n = 10/group) and the 50 μM metformin group (n = 10/group) after 3 days were observed under conditions of 14 h of light and 10 h of alternating darkness (LD). (k) The behavioral rhythms of the Con group (n = 10/group) and Met group (n = 10/group) were observed on day 3 under continuous light (LL) conditions. (l) The 30 min behavioral rhythms of the Con group (n = 10/group) and Met group (n = 10/group) were observed under continuous light (LL) and continuous darkness (LD) conditions. (**** p < 0.0001, *** p < 0.001, ** p < 0.01, “ns”, no statistical difference).
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