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. 2018 Jul 3;7:e33910. doi: 10.7554/eLife.33910

Figure 5. DLK inhibition prevents DLK-dependent gene upregulation, mechanical allodynia, and microgliosis after SNI.

Figure 5.

(A–C) GNE-3511 treatment prevents upregulation of DLK-dependent genes in the DRG after SNI. (A) Representative images of Atf3 (green), Csf1 (orange), and Tubb3 (red) in situ hybridizations in ipsilateral L4 DRGs from vehicle-treated, GNE3511-treated, and DLK cKO mice 5 days post-SNI. Scale bar 100 μm. (B, C) Quantification of neuronal gene expression intensity in ipsilateral L4 DRGs at 5 dpi showing reduced expression of Atf3 (B) and Csf1 (C) in DLK cKO and GNE-3511 vs vehicle treated mice (n = 2 per group). *p<0.05, **p<0.01 by Tukey’s multiple comparisons test. (D) DLK inhibitor study design. vF: von Frey; BL: baseline; SNI: spared nerve injury; 1 w: 1 week post SNI; TH: tissue harvest. Twice daily dosing of GNE-3511 began at 18 hr post injury (hpi). (E) DLK inhibitor GNE-3511 prevents mechanical allodynia measured 1 week post injury. Vehicle n = 12; GNE-3511 n = 17. ***p=0.0004 for genotype by 2-way ANOVA. (F–H) GNE-3511 prevents the spinal cord microgliosis elicited by SNI as shown by Iba1 staining in cleared immunostained lumbar spinal cord. Representative images from vehicle (F) vs GNE-3511 treated mice (G) and quantification of Iba1-positive signal in ipsilateral spinal cord (H). Vehicle n = 4; GNE-3511 n = 7. ***p=0.0001 by 2-tailed Student’s t test. Scale bar in (F), (G) 400 µm.

Figure 5—source data 1. data plotted in Figure 5B,C; quantification of in situ signal intensity at 5 dpi for Atf3 and Csf1 in ipsilateral and contralateral L4 DRGs normalized by total neuronal volume sampled as labeled by Tubb3.
elife-33910-fig5-data1.xlsx (59.2KB, xlsx)
DOI: 10.7554/eLife.33910.027
Figure 5—source data 2. data plotted in Figure 5E; von Frey thresholds (>5 responses out of 10 stimulations) in vehicle vs GNE-3511-treated at 7 dpi.
elife-33910-fig5-data2.xlsx (36.3KB, xlsx)
DOI: 10.7554/eLife.33910.028
Figure 5—source data 3. data plotted in Figure 5H; quantification of Iba1-positive signal in vehicle vs GNE-3511-treated dorsal spinal cord ipsilateral to injury at 8 dpi.
elife-33910-fig5-data3.xlsx (29.4KB, xlsx)
DOI: 10.7554/eLife.33910.029