Fig. 3: CRMP5 autoimmunity induces mechanical allodynia and hyperexcitability in both male and female rats.

Rats were immunized by three injections (indicated by arrows) of 50 µg of pCMV2 plasmid, allowing for the expression of CRMP5 or control (empty) in the spinodeltoidus muscle. Rats received an intramuscular injection of a plasmid (pCMV2-CRMP5) carrying the coding sequence for CRMP5 on day 0 and then two booster shots at weeks 2 and 4 after the first injection. Graph showing the paw withdrawal threshold of A male and E female rats injected with the CRMP5 coding plasmid compared to the control empty plasmid (n = 8 animals per group; *p < 0.05, two-way ANOVA). Bar graph with scatter plot showing the area under the curve for the mechanical thresholds in B male and F female rats injected as described above (n = 8 animals per group; *p < 0.05, two-tailed, Mann–Whitney test). Rats were tested for their thermal thresholds using Hargreave’s test, and no difference was found in C male and G female rats injected with the CRMP5 coding plasmid compared to the control empty plasmid (n = 8 animals per group; two-way ANOVA). Bar graph with scatter plot showing the area under the curve for the thermal thresholds in D male and H female rats injected as described above (n = 8 animals per group; two-tailed, Mann–Whitney test. I Representative recordings in response to a depolarizing current step to evoke action potentials (APs) in sensory neurons from male and female rats injected with a plasmid expressing CRMP5 or a control plasmid. J Summary of the number of APs in the indicated conditions. *p < 0.05, multiple Mann–Whitney tests. Male control n = 10 cells, Male CRMP5 n = 7 cells, Female control n = 11 cells, Female control n = 10 cells from at least three rats. K Representative recordings in response to various steps of depolarizing current to measure rheobase in sensory neurons prepared from rats with CRMP5 autoimmunity or control. L Summary of the measured rheobase in indicated conditions. Asterisks indicate significance compared with control *p < 0.05, two-tailed, Mann–Whitney test. Male control n = 10 cells, Male CRMP5 n = 7 cells, Female control n = 11 cells, Female control n = 10 cells from at least three rats. All data are shown with error bars that indicate mean ± SEM. See Supplementary Data 1 for additional statistical details. Source data are provided as a Source Data file.