Figure 1.
Glycolysis, but not mitochondrial respiration, is necessary to provide ATP for axonal retraction. (A) Image series of video-microscopy showing the dynamics of axonal tip position before and during WIN+metabolic drugs exposure in media containing 10 mM glucose and 1 mM pyruvate. Hippocampal neurons expressing LifeAct-mCherry were imaged at a frequency of 0.5 frames/min for 30 min before the induction of retraction. Oligomycin, 2DG, or DMSO (control) were added in combination with WIN. On the top right is shown the quantification of the position of the growth cone (distance from position relative to time zero corresponding to the injection of WIN). On the bottom right, the boxplot shows values 20 min after drug injections; 51 < n < 58 axons from three independent experiments. (B) Image series of video-microscopy showing the dynamics of axonal tip position before and during WIN exposure in media containing 10 mM glucose or 1 mM pyruvate in hippocampal neurons. On the right, the boxplot shows values 20 min after drug injections; 43 < n < 94 axons from two independent experiments. (C) Boxplot showing values 20 min after WIN injection and 40 min after treatment with DMSO (control; n = 19, two independent experiments), 5 mM sodium azide (SA; n = 30, three independent experiments), 1 mM iodoacetate (IA; n = 25; two independent experiments), or 30 mM 2DG (n = 38, two independent experiments) in hippocampal neurons. (D) Boxplot showing values 20 min after 1 µM 2-AG injection in combination with oligomycin (oli), 2DG; or DMSO in hippocampal neurons; 14 < n < 77 axons from two independent experiments. (E) Boxplot showing values 20 min after 2 µM LPI injection in combination with oligomycin, 2DG, or DMSO in cortical neurons; 13 < n < 41 axons from two independent experiments. (F) Boxplot showing values 20 min after 0.2 U/ml calpeptin in combination with oligomycin, 2DG, or DMSO in cortical neurons; 52 < n < 101 axons from two independent experiments. For all experiments, cell concentrations of the drugs were 100 nM WIN, 10 µM oligomycin, and 30 mM 2DG. Statistics: one-way ANOVA followed by Tukey HSD post-test correction; ***P < 0.001.

Glycolysis, but not mitochondrial respiration, is necessary to provide ATP for axonal retraction. (A) Image series of video-microscopy showing the dynamics of axonal tip position before and during WIN+metabolic drugs exposure in media containing 10 mM glucose and 1 mM pyruvate. Hippocampal neurons expressing LifeAct-mCherry were imaged at a frequency of 0.5 frames/min for 30 min before the induction of retraction. Oligomycin, 2DG, or DMSO (control) were added in combination with WIN. On the top right is shown the quantification of the position of the growth cone (distance from position relative to time zero corresponding to the injection of WIN). On the bottom right, the boxplot shows values 20 min after drug injections; 51 < n < 58 axons from three independent experiments. (B) Image series of video-microscopy showing the dynamics of axonal tip position before and during WIN exposure in media containing 10 mM glucose or 1 mM pyruvate in hippocampal neurons. On the right, the boxplot shows values 20 min after drug injections; 43 < n < 94 axons from two independent experiments. (C) Boxplot showing values 20 min after WIN injection and 40 min after treatment with DMSO (control; n = 19, two independent experiments), 5 mM sodium azide (SA; n = 30, three independent experiments), 1 mM iodoacetate (IA; n = 25; two independent experiments), or 30 mM 2DG (n = 38, two independent experiments) in hippocampal neurons. (D) Boxplot showing values 20 min after 1 µM 2-AG injection in combination with oligomycin (oli), 2DG; or DMSO in hippocampal neurons; 14 < n < 77 axons from two independent experiments. (E) Boxplot showing values 20 min after 2 µM LPI injection in combination with oligomycin, 2DG, or DMSO in cortical neurons; 13 < n < 41 axons from two independent experiments. (F) Boxplot showing values 20 min after 0.2 U/ml calpeptin in combination with oligomycin, 2DG, or DMSO in cortical neurons; 52 < n < 101 axons from two independent experiments. For all experiments, cell concentrations of the drugs were 100 nM WIN, 10 µM oligomycin, and 30 mM 2DG. Statistics: one-way ANOVA followed by Tukey HSD post-test correction; ***P < 0.001.

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