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. 2019 May 16;13:209. doi: 10.3389/fncel.2019.00209

FIGURE 4.

FIGURE 4

Pridopidine and S1R agonist 3-PPP both restore homeostatic plasticity at a 1 μM concentration. (A) Representative traces of mEPSC recordings from WT CPNs treated with TTX and/or 1 μM pridopidine. (B) mEPSC amplitude of WT CPNs treated with TTX and/or 1 μM pridopidine. Cells treated with only TTX had larger mEPSCs than those treated with only vehicle control (∗∗p < 0.01, Bonferroni post hoc t-test), and cells treated with TTX and pridopidine had larger mEPSCs than those treated with only pridopidine (∗∗p < 0.01, Bonferroni post hoc t-test; TTX p < 0.001, two-way ANOVA). (C) mEPSC frequency of WT CPNs treated with TTX and/or 1 μM pridopidine. Cells treated with only TTX had more frequency mEPSCs than those treated with only vehicle control (∗∗∗p < 0.001, Bonferroni post hoc t-test), while those treated with both TTX and pridopidine showed a non-significant trend toward more frequent mEPSCs than those treated with only pridopidine (p = 0.06, Bonferroni post hoc t-test; TTX p < 0.001, two-way ANOVA). For both (B) and (C), cells per group n = 24, culture batch N = 4. (D) Representative traces of mEPSC recordings from YAC128 CPNs treated with TTX and/or 1 μM pridopidine. (E) mEPSC amplitude of YAC128 CPNs treated with TTX and/or 1 μM pridopidine. Cells treated with both TTX and pridopidine had higher amplitude mEPSCs than those treated with only TTX (∗p < 0.05, Bonferroni post hoc t-test), or pridopidine (∗∗∗p < 0.001, Bonferroni post hoc t-test; TTX p < 0.01, TTX/pridopidine interaction p < 0.001, two-way ANOVA). (F) mEPSC frequency of YAC128 CPNs treated with TTX and/or 1 μM pridopidine. Cells treated with both TTX and pridopidine had more frequent mEPSCs than those treated with pridopidine alone (∗∗p < 0.01, Bonferroni post hoc t-test; TTX p < 0.05, TTX/pridopidine interaction p < 0.05, two-way ANOVA). For (E) and (F), control n = 22, TTX n = 25, pridopidine n = 24, TTX + pridopidine n = 27, culture batch N = 4. (G) Representative traces of mEPSC recordings from YAC128 CPNs treated with TTX and/or 3-PPP. (H) mEPSC amplitude from YAC128 CPNs treated with TTX and/or 3-PPP. Cells treated with both 3-PPP and TTX had larger mEPSCs than those treated with only 3-PPP (∗p < 0.05, Bonferroni post hoc t-test; TTX p < 0.05, two-way ANOVA). (I) mEPSC frequency from YAC128 CPNs treated with TTX and/or 3-PPP. Cells treated with both 3-PPP and TTX had more frequent mEPSCs than those treated with only 3-PPP (∗p < 0.05, Bonferroni post hoc t-test; TTX/3-PPP interaction p < 0.05, TTX p = 0.09, two-way ANOVA).; For (H) and (I), control n = 14, TTX n = 13, 3-PPP n = 14, TTX/3-PPP n = 13, culture batch N = 12. (J) Representative traces of mEPSC recordings from YAC128 CPNs treated with TTX and/or 100 nM pridopidine. (K) mEPSC amplitude of YAC128 CPNs treated with TTX and/or 100 nM pridopidine. YAC128 CPNs treated with both TTX and pridopidine had higher mEPSC amplitude than those treated with only pridopidine (∗p < 0.05, Bonferroni post hoc t-test; TTX p < 0.01, two-way ANOVA). (L) mEPSC frequency of YAC128 CPNs treated with TTX and/or 100 nM pridopidine. There was no difference in mEPSC frequency between any groups (control vs. TTX: p > 0.99; pridopidine vs. pridopidine+TTX: p > 0.99; control vs. pridopidine: p = 0.83; TTX vs. pridopidine+TTX: p = 0.56, Bonferroni post hoc t-test). For (K) and (L), control n = 19, TTX n = 20, pridopidine n = 21, TTX + pridopidine n = 23, culture batch N = 4. (M) Representative traces of mEPSC recordings from YAC128 CPNs treated with TTX and/or 10 μM pridopidine. (N) mEPSC amplitude of YAC128 CPNs treated with TTX and/or 10 μM pridopidine. Cells treated with both TTX and pridopidine had larger mEPSCs than those treated with only pridopidine (∗p < 0.05, Bonferroni post hoc t-test; TTX p < 0.01, two-way ANOVA). (O) mEPSC frequency of YAC128 CPNs treated with TTX and/or 10 μM pridopidine. There was no difference in frequency between any groups (control vs. TTX: p = 0.92; pridopidine vs. pridopidine+TTX: p = 0.18; control vs. pridopidine: p = 0.72; TTX vs. pridopidine+TTX: p > 0.99, Bonferroni post hoc t-test). For (N) and (O), control n = 20, TTX n = 22, pridopidine n = 21, TTX + pridopidine = 19; culture batch N = 4.