D-Aspartate treatment attenuates myelin damage and stimulates myelin repair (2018)
Figure 4

Figure 4. Contribution of NMDA receptor and NCX3 exchanger activities on D-Asp-induced [Ca2+]i levels in oligodendrocyte precursors.

A-B. left panels. Superimposed single-cell traces representative of the effect of 100 μM D-Asp on [Ca2+]i detected in MO3.13 cells (A) and primary OPC (B) in absence or in presence of 10 μM MK-801 or 150 μM APV. A-B. right panels. Quantification of the oscillation index in MO3.13 cells (A) and primary OPC (B) in absence or in presence of 10 μM MK-801 or 150 μM APV. a-b, Quantification of the initial [Ca2+]i increase elicited by D-Asp measured as ∆% of peak versus basal values in absence or in presence of 10 μM MK-801 or 150 μM APV, both in M03.13 cells (a) and primary OPC (b). MK-801 and APV were preincubated 10 minutes before registration.

C-D. left panels; Superimposed single-cell traces representative of the effect of 100 μM D-Asp on [Ca2+]i detected in MO3.13 cells (C) and primary OPC (D) in absence or in presence of 30 nM YM-244769 or 100 nM BED. C-D, right panels; Quantification of the oscillation index in MO3.13 cells (C) and primary OPC (D) in absence or in presence of 30 nM YM-244769 or 100 nM BED. c-d, Quantification of the initial [Ca2+]i increase elicited by D-Asp measured as ∆% of peak versus basal values in absence or in presence of 30 nM YM-244769 or 100 nM BED, both in M03.13 cells (c) and primary OPC (d). YM-244769 or BED were preincubated 10 minutes before registration.

E. left panel; Superimposed single-cell traces representative of the effect of 100 μM D-Asp on [Ca2+]i detected in MO3.13 cells in presence of siCtl or sincx3 silencing. E, right panel; Quantification of the oscillation index in MO3.13 cells in absence or in presence of sincx3. e, Quantification of the initial [Ca2+]i increase elicited by D-Asp and measured as ∆% of peak versus basal values in absence or in presence of sincx3.

F. left panel. Superimposed single-cell traces representative of the effect of 100 μM D-Asp on [Ca2+]i detected in primary OPC obtained from wild-type ncx3+/+, heterozygous ncx3 +/-, and knock-out ncx3-/- mice. F. right panel. Quantification of the oscillation index elicited by D-Asp in primary mouse OPC obtained from ncx3+/+, ncx3 +/-, and ncx3-/- mice. f, Quantification of the initial [Ca2+]i increase measured as ∆% of peak versus basal values.

Data information: The values represent the mean ± S.E.M from 3 independent experimental sessions. Level of significance was determined by using: in A and a, one way-ANOVA P<0.001 followed by Bonferroni post hoc test, *P<0.05 versus control (basal value), ˄P<0.05 versus D-Asp. Data are reported as mean of 23-30 cells in each group, n=3 biological replicates; B and b, one way-ANOVA, P<0.001 followed by Bonferroni post hoc test, *P<0.05 versus control (basal value), ˄P< 0.05 versus D-Asp. Data are reported as mean of 20-23 cells in each group, n=3 biological replicates; C and c, one way-ANOVA P<0.001 followed by Bonferroni post hoc test, *p< 0.05 versus control (basal value), ˄ P< 0.05 versus D-Asp. Data are reported as mean of 19-30 cells in each group, n=3 biological replicates; D and d, one way-ANOVA P<0.001 followed by Bonferroni post hoc test, *P< 0.05 versus control (basal value), ˄P< 0.05 versus D-Asp. Data are reported as mean of 18-20 cells in each group, n=3 biological replicates; E and e, one way-ANOVA P<0.001 followed by Bonferroni post hoc test, *P< 0.05 versus sictl, ˄P< 0.05 versus D-Asp + sictl. Data are reported as mean of 25-30 cells in each group, n=3 biological replicates; F and f, one way-ANOVA P<0.001 and P=0.003, respectively, followed by Bonferroni post hoc test, *P< 0.05 versus basal value, ˄P< 0.05 versus ncx3+/+ and ncx3+/-. Data are reported as mean of 10-19 cells in each group, n=3 biological replicates. See the exact P‐values from comparisons tests in Appendix Table S4.