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. 2005 Aug 1;170(3):349–355. doi: 10.1083/jcb.200504028

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Copyright © 2005, The Rockefeller University Press

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Figure 3. — Exogenously provided NAD and nicotinamide delay Wallerian degeneration in soma-free axonal segments. (A) Wallerian degeneration of soma-separated axonal segments is delayed by NAD and nicotinamide (NAm) without pretreatment. DRG explants cultured for 5 d were subjected to axonal transection and the ganglia were removed. At the same time, indicated concentrations of NAD or nicotinamide were added to the medium of the axonal segments only. Quantification of axon degeneration was performed from phase images taken at different time-points after transection. Statistical analysis was done by two-way ANOVA using GraphPad Prism software. P < 0.001 for NAD (5 mM, 20 mM), NAm (5 mM, 25 mM) vs. control. (B and C) Quantification of the protective effects of 1 mM (B) and 5 mM (C) NAD added at indicated time-points. Pre 24h: NAD added 24 h before transection; pre 0h: NAD added at the same time as transection; post 3, 5, 7 h: NAD added 3, 5, and 7 h post-transection, respectively. Quantification was performed from phase images taken at indicated time-points (B) or at 12 h after transection (C). Statistical analysis: two-way ANOVA in B: P < 0.001 for NAD 1 mM pre 0 h and 1 mM pre 24 h vs. control; P > 0.05 for NAD 1 mM pre 0 h vs. pre 24 h; t test in C: ***, P < 0.001. (D) Nicotinamide (NAm) attenuates NAD decrease in soma-free axonal segments. DRG explants cultured for 5 d were subjected to axonal transection and the ganglia were removed. At the same time, indicated concentrations of nicotinamide were added to the medium with axonal segments only. The axonal samples were collected at indicated time-points after transection and subjected to HPLC analysis. The results from three independent experiments are presented and statistical analysis was done by two-way ANOVA using GraphPad Prism software. P < 0.001 for 5 mM NAm and 25 mM NAm vs. control.