Journal: iScience

Article Title: Successful axonal regeneration is associated with intraneuronal metabolic reprogramming

doi: 10.1016/j.isci.2025.113631

Figure Lengend Snippet: Inhibiting glycolysis significantly reduces axonal regeneration in adult zebrafish retinal explants (A) Schematic overview of the experimental procedure. Retinas from naive Tg(Tru.gap43:eGFP) mil1 adult zebrafish were dissected, cut into explants, and cultured, as described previously. After four days, regenerated axons were quantified. (B) Six glycolysis inhibition conditions were tested: (1) 2-DG (2-deoxy-D-glucose, orange); (2) lactate (Lac, green); (3) Galactose (Gal, red); (4) an inhibitor mix (purple, 9 μM of 3PO, 9 μM of Malonyl-NAC, and 9 μM of GSK2837808A (GSK)); (5) 9 μM of 3PO alone (brown); (6) 9 μM of GSK alone (pink). (C) Representative images show that 2-DG caused the most severe axonal outgrowth reduction, with extremely few axons growing only short distances. Similarly, lactate treatment severely reduced axonal outgrowth, resulting in a limited number of regenerating axons. Galactose and inhibitor mix treatments also significantly impaired axonal outgrowth, but less severely. 3PO or GSK treatment induced a moderate reduction in regrowing axons compared to the vehicle. Scale bar 200 μm. (D) Cumulative distribution of axonal density profiles over distance from the explant. The 2-DG and lactate treatment severely impaired both the density and length of axons. Galactose or inhibitor mix administration caused a reduction in both axonal sprouting and elongation, but less severe than 2-DG and lactate. Treatments with 3PO or GSK induced slightly reduced sprouting, but strongly impaired elongation. See also . (E) Quantification of cumulative axonal density revealed a ∼90% reduction with 2-DG treatment, ∼85% with lactate, ∼60% with galactose or the inhibitor mix, and ∼40% with 3PO or GSK, compared to the vehicle control. Data of each condition were collected from N = 13–23 explants, derived from n = 26 fish across eight independent experiments, presented as median ±95% confidence interval (D) or median ±SEM and bootstrap 95% confidence interval versus vehicle-treated fish (E). One-way Kruskal-Wallis ANOVA (E). p values are reported within graphs. Abbreviations: DIV (day in vitro ), Veh (vehicle). Figure containing assets from BioRender.com .

Article Snippet: GSK2837808A , MedChemExpress , Cat# HY-100681.

Techniques: Cell Culture, Inhibition, Control, Derivative Assay, In Vitro