Fig 2: BDNF and TrkB in the L4-L6 SCDH plays a key role in the hyperalgesic priming model. (A-B) The quantification of the Western blots results and a representative Western blot showing BDNF and p-TrkB protein isolated from the ipsilateral SCDH 48 h after 2nd injection. (C) Representative immunofluorescence images of BDNF (red) merged with p-TrkB (green) in ipsilateral L4-6 SCDH of hyperalgesic priming rats. (D-F) The immunofluorescence quantification of BDNF and p-TrkB co-expression cells in ipsilateral L4-6 SCDH. (G-H) Evaluation of BDNF and p-TrkB gene expression in ipsilateral L4-6 SCDH using qPCR in the sHP, HP, HP + EA, HP + sEA groups 48 h after 2nd injection. Data are presented as mean ± SEM, n = 5; *p < 0.05, **p < 0.01 vs sHP group, #p < 0.05, ##p < 0.01 vs HP group. Two-way ANOVA with Tukey's post-hoc test was applied in. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Fig 3: BDNF neutralizing antibody intervention reduced phosphorylation of TrkB. (A-B) The quantification of the Western blots results and a representative Western blot showing BDNF and p-TrkB protein isolated from the ipsilateral SCDH 48 h after 2nd injection. (C) Representative immunofluorescence images of BDNF (red) merged with p-TrkB (green) in ipsilateral L4-6 SCDH of BDNF neutralizing antibody intervention rats. (D-F) The immunofluorescence quantification of BDNF and p-TrkB co-expression cells in ipsilateral L4-6 SCDH. (G-H) Evaluation of BDNF and p-TrkB gene expression in ipsilateral L4-6 SCDH using qPCR in the sHP + IgG, HP + IgG, HP + BDNF neutralizing antibody groups 48 h after 2nd injection. Data are presented as mean ± SEM, n = 5; *p < 0.05, **p < 0.01 vs sHP + IgG group, #p < 0.05, ##p < 0.01 vs HP + IgG group. Two-way ANOVA with Tukey's post-hoc test was applied in. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Fig 4: Acute effects of TC-G 1008 (TC-G) (20 mg/kg), ZnCl2 (16 mg Zn/kg), or VPA (150 mg/kg) and MES seizures on the relative expression of proteins: phosphorylated CREB (p-CREB), total CREB, BDNF, phosphorylated TrkB (p-TrkB) and TrkB in the hippocampi of mice fed the ZnA or ZnD diet. Hippocampal samples were obtained ca. 3 min after MES. (A–F) Data are expressed as means ± SEM of p-CREB/CREB or BDNF/ß-actin or p-TrkB/TrkB ratio. They were analyzed by one-way ANOVA. n = 4–6 in each group. Statistical details: (A) F(4,23) = 1.100, p = 0.3805, (B) F(4,23) = 0.6353, p = 0.6425, (C) F(4,23) = 0.5934, p = 0.6709, (D) F(4,23) = 1.218, p = 0.3306, (E) F(4,23) = 1.100, p = 0.3805, (F) F(4,23) = 0.6353, p = 0.6425 (A-F one-way ANOVA). (G) Representative blots of p-CREB (Ser 133), CREB (~46 kDa), BDNF (~14 kDa), p-TrkB (Tyr 816), TrkB (~140 kDa) and ß-actin (~42 kDa) in the hippocampi of mice. #p-CREB and p-TrkB come from the same blot, thus sharing the corresponding ß-actin band.

Fig 5: BDNF recombinant protein intervention increased phosphorylation of TrkB. (A-B) The quantification of the Western blots results and a representative Western blot showing BDNF and p-TrkB protein isolated from the ipsilateral SCDH 48 h after 2nd injection. (C) Representative immunofluorescence images of BDNF (red) merged with p-TrkB (green) in ipsilateral L4-6 SCDH of BDNF recombinant protein intervention rats. (D-F) The immunofluorescence quantification of BDNF and p-TrkB co-expression cells in ipsilateral L4-6 SCDH. (G-H) Evaluation of BDNF and p-TrkB gene expression in ipsilateral L4-6 SCDH using qPCR in the HP + PBS, HP + EA + PBS, HP + EA + BDNF groups 48 h after 2nd injection. Data are presented as mean ± SEM, n = 5; *p < 0.05, **p < 0.01 vs HP + PBS group, #p < 0.05, ##p < 0.01 vs HP + EA + PBS group. Two-way ANOVA with Tukey's post-hoc test was applied in. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)