Fig. 4

Fig. 4 Endogenous mGluR5 is required for CHPG-induced high-amplitude myelin Ca²⁺ transients to occur.

a, Schematic of treatment of larvae and live imaging of Ca²⁺ activity in myelin sheaths. b,b′, Top shows representative maximum time series projections of Tg(mbp:memGCaMP7s) fluorescence with Ca²⁺ transients detected by AQuA2 false-colored automatically; bottom show single time-point images from the time-lapse series and highlight transients identified within yellow boxes. WT (N = 14) (b) and grm5a^−/−grm5b^−/− animals (b′) (N = 16) treated with vehicle and CHPG (WT N = 15; grm5a^−/−grm5b^−/−N = 17). Scale bar, 10 µm. c, Median amplitude (dF/F) of Ca²⁺ transients per fish (one-way ANOVA; P = 0.0079; Tukey’s multiple comparison test; WT DMSO versus CHPG P = 0.111, WT DMSO versus grm5a^−/−grm5b^−/− DMSO P = 0.9633; WT DMSO versus grm5a^−/−grm5b^−/− CHPG P = 0.9210, WT CHPG versus grm5a^−/−grm5b^−/− DMSO P = 0.0310, WT CHPG versus grm5a^−/−grm5b^−/− CHPG P = 0.0407; grm5a^−/−grm5b^−/− CHPG versus grm5a^−/−grm5b^−/− DMSO P = 0.9987) (WT DMSO N = 14, 0.94 ± 0.24, WT CHPG N = 15, 1.21 ± 0.31, grm5a^−/−grm5b^−/− DMSO N = 16, 0.98 ± 0.16, grm5a^−/−grm5b^−/− CHPG N = 17, 0.99 ± 0.19). d, Frequency of myelin Ca²⁺ transients in CHPG and DMSO-treated grm5a^−/−grm5b^−/− mutant and WT animals (one-way ANOVA P = 0.1658; Holm–Šídák’s multiple comparisons tests: WT DMSO versus WT CHPG P = 0.3022; WT DMSO versus grm5a^−/−grm5b^−/− DMSO P = 0.2591; WT CHPG versus grm5a^−/−grm5b^−/− DMSO P = 0.9840; WT CHPG versus grm5a^−/−grm5b^−/− CHPG P = 0.6868; WT DMSO versus grm5a^−/−grm5b^−/− CHPG P = 0.6868). WT DMSO 138 ± 149.2, WT CHPG 74.39 ± 48.68, grm5a^−/−grm5b^-/, DMSO 75.04 ± 82.95, grm5a^−/−grm5b^−/− CHPG 108.9 ± 95.43. Scale bar, 10 µm. e, Median duration of Ca²⁺ transients per fish (one-way ANOVA P = 0.2687; Tukey’s multiple comparisons test: WT DMSO versus WT CHPG P = 0.4745; WT DMSO versus grm5a^−/−grm5b^−/− DMSO P = 0.9864; WT DMSO versus grm5a^−/−grm5b^−/− CHPG P = 0.8633; WT CHPG versus grm5a^−/−grm5b^−/− DMSO P = 0.2638, WT CHPG versus grm5a^−/−grm5b^−/− CHPG P = 0.08864; grm5a^−/−grm5b^−/− DMSO versus grm5a^−/−grm5b^−/− CHPG P = 0.6516). WT DMSO 12.76 ± 6.964, WT CHPG 9.495 ± 5.044, grm5a^−/−grm5b^−/−, DMSO 13.52 ± 5.936, grm5a^−/−grm5b^−/−, CHPG 11.06 ± 6.233. f, Ca²⁺ transient dF/F traces of the events shown in corresponding conditions in b and b'. Data show mean ± s.d.

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