FIGURE 2.

Determination of specific NR2B labeling. To determine the specific NR2B labeling, DRG sections were incubated with anti-NR2B (A) and three negative controls were used, one in which the primary antibody (anti-NR2B) was omitted (B), another in which the primary antibody was replaced with goat IgG (C), and the third one in which the primary antibody was pre-incubated with blocking peptide (D). (A–D) Show four panels: top left corresponds to the red signal: NR2B-labeling + background (A) or only background (B-D). Top right: Nissl staining. Bottom left: light picture. Bottom right: overlap of the three panels. (E) Shows intensity labeling in the red channel for the four conditions. Each dot represents a measurement from a single soma, red lines indicate the mean ± SD. Based on this result we decided to use the negative IgG goat group to set the background value, which consisted in the average value plus one standard deviation. Specific NR2B intensity labeling (NR2B labeling) was defined as: mean value of the “Positive group” minus the mean value + standard deviation of the “negative IgG group” (indicated with the bracket). (F) Shows that the “NR2B labeling” for the neuronal soma was the same for Naïve and Sham rats (mean ± SEM, normalized to the Naïve values). No significant difference was found between any of the groups (One-way ANOVA). The same was observed for SGCs and perinuclear areas between Naïve and Sham rats (not shown). (G) The sural and tibial nerves innervate the lateral and middle areas, respectively. In sural-spared nerve injury (SNI) the sural area is spared and the tibial-area in non-spared. In tibial-SNI the tibial area is spared and the sural area in non-spared. Rectangle indicate the non-spared regions and ovals the spared regions for each of the SNI variants.