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. 2016 Dec 12;213(13):2949–2966. doi: 10.1084/jem.20160877

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© 2016 Hayano et al.

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Figure 6. — The enhanced excitatory synaptic transmission in the model of neuropathic pain is abolished by a Netrin-4–blocking antibody. (A and B) Penetration of the Netrin-4 antibody (Ab) into the dorsal horn. (B) Double immunofluorescence staining for the fluorescent dye–conjugated Netrin-4 antibody (HL-555; red) and NeuN (green). The fluorescence of the antibody was observed on neurons (arrowheads) and also around blood vessels (arrow) in lamina III (dashed lines in A). Bars: (A) 0.2 mm; (B) 10 µm. (C) Paw withdrawal of the control IgG (closed; n = 11)- or Netrin-4 antibody (open; n = 11)–treated animals after PSL injury. The data are presented as the mean ± SEM. *, P < 0.05; **, P < 0.01; Tukey-Kramer test. (D) Representative traces of the sEPSCs in the contralateral side, ipsilateral side, control IgG-treated, and Netrin-4 antibody–treated spinal cords. Electrophysiological data represent the mean from 25 slices. (E and F) The frequency (E) and amplitude (F) of the sEPSCs in each group. *, P < 0.05 vs. the contralateral side or control IgG-treated spinal cords; Student’s t test. (G) Representative traces of the eEPSCs recorded after stimulation of a neighboring area. (H) The I-O curve of eEPSCs. *, P < 0.05; ***, P < 0.001 vs. the contralateral side. ^#, P < 0.05; ^###, P < 0.001 vs. the control IgG-treated spinal cord; Bonferroni test. (I) Representative traces of non-NMDAR–mediated eEPSCs at −60 mV and NMDAR-mediated eEPSCs at 60 mV. (J) The ratio of NMDAR/non-NMDAR–mediated components of the eEPSCs in each group. **, P < 0.01 vs. the contralateral side; ***, P < 0.001 vs. the control IgG-treated spinal cord; Student’s t test. The data are presented as the mean ± SEM. Each experiment was performed twice in A and B and three times in C–J. contra, contralateral; ipsi, ipsilateral.