We analyzed levels of p-STAT3 in the proximal nerve stump 1 day after sciatic nerve lesion. In WT, p-STAT3 is barely detectable in the unlesioned contralateral learn more nerve but is dramatically upregulated by injury (Figure 3C). p-STAT3 is localized in neuronal axons, as shown by immunostaining (Figure 3D). In the absence of DLK, STAT3 is still phosphorylated in the injured axons, and the levels

are similar to WT (Figures 3C and 3D; n = 3). These data show that the local activation of STAT3 does not require DLK. Instead, these findings suggest that DLK may be necessary for translocation of the injury signal to the cell body. We next examined whether DLK is indeed required for the transport of p-STAT3 to the cell body. To track the movement of the phosphorylated protein upon injury, we performed a double nerve ligation in which the sciatic nerve is sutured at two locations

(Figure 3E). The nerve ligation injures axons and blocks axonal transport, so that transported cargoes accumulate near the knots. Retrograde cargoes accumulate in the proximal segment of the nerve, while anterograde cargoes concentrate in the distal segment, so the ratio of protein present in the proximal/distal segment is a measure selleck inhibitor of retrograde transport (Cavalli et al., 2005). Upon double ligation of WT sciatic nerves for 6 hr, p-STAT3 levels are 1.5-fold higher in the proximal segment, consistent with the retrograde transport of p-STAT3 after injury. However, this accumulation is blocked in DLK KOs (p < 0.05) (Figures 3E and 3F). We also analyzed transport of JIP3, a scaffolding protein that links DLK and JNK to the axon transport machinery (Cavalli et al., 2005; Ghosh et al., 2011). Injury facilitates the association of

JIP3 with the retrograde transport machinery and increases the retrograde transport of both JIP3 and phosphorylated JNK (Cavalli et al., Astemizole 2005). In the double ligation assay, injury-induced accumulation of JIP3 in the proximal stump is abolished in the absence of DLK (p < 0.05) (Figures 3E and 3F). Therefore, DLK is necessary for the retrograde transport of both p-STAT3 and JIP3 upon axon injury. Collectively, these results demonstrate that DLK plays an essential role for the axonal transport of injury signaling components to the cell body. Taken together, these data demonstrate that DLK is required for robust axon regeneration in the vertebrate PNS in vivo, DLK promotes retrograde transport of injury signals that enhance axonal regenerative capacity, and injury-induced potentiation of axonal regeneration requires DLK. Trauma, neurotoxins, and neurological disease can all trigger axonal damage and the loss of neuronal connections. The capacity of a neuron to regenerate an injured axon is crucial for the recovery of neural function.