Extended Data Fig. 8: mGluR5 regulates myelin Ca2+ levels and myelin volume.
From: Activity-driven myelin sheath growth is mediated by mGluR5
Whole CNS measurement of mbp:memGCaMP7s fluorescent intensity and myelin volume using computational 3D reconstruction of SDCM images. A: Representative image of a 4 dpf zebrafish expressing mbp:memGCaMP7s in spinal oligodendrocytes and Schwann cells. Scale bar,100 µm. B: Computational reconstruction of the transgenic myelin reporter mbp:memGCaMP7s used to assess myelin Ca2+ baseline activity and myelin volume. C: Reconstruction of 4 dpf CNS myelin in animals treated with DMSO (N = 17), MTEP (N = 28) and CHPG (N = 20). Green reflects myelin along dorsal axons, while yellow labels ventral myelin. D: Doral myelin as volumetric pixel volume (vpx) (One-was ANOVA p-value = <0.0001; Tukey’s multiple Comparisons test: DMSO vs. CHPG p-value = 0.0194; DMSO vs. MTEP p-value = 0.0090; CHPG vs. MTEP p-value = <0.0001) DMSO: mean: 337125, SD: ±51246; MTEP mean: 271853, SD: ±60709; CHPG mean: 400665, SD: ±90675). E: Relative Fluorescent intensity (RFU) in dorsal myelin sheaths of mbp:memGCaMP7s, reflecting baseline myelin Ca2+ activity (One-was ANOVA p-value = <0.0001; Tukey’s multiple Comparisons test: DMSO vs. CHPG p-value = 0.0344; DMSO vs. MTEP p-value = 0.0068; CHPG vs. MTEP p-value = <0.0001; DMSO: mean: 54580775 SD: ±13046833; MTEP mean: 39357143, SD: ±21823431; CHPG mean: 67789985, SD: ±21823431). F: Ventral myelin as volumetric pixel volume (vpx) (One-was ANOVA p-value = <0.0001; Tukey’s multiple Comparisons test: DMSO vs. CHPG p-value = 0.0398; DMSO vs. MTEP p-value = 0.0131; CHPG vs. MTEP p-value = <0.0001). DMSO: mean: 593429, SD: ±146357; MTEP mean: 484280, SD: ±111349; CHPG mean: 694801, SD: ±116964). G: Relative Fluorescent intensity (RFU) in ventral myelin sheaths of mbp:memGCaMP7s, reflecting baseline myelin Ca2+ activity (One-was ANOVA p-value = <0.0001; Tukey’s multiple Comparisons test: DMSO vs. CHPG p-value = 0.0030; DMSO vs. MTEP p-value = 0.1009; CHPG vs. MTEP p-value = <0.0001). DMSO: mean: 198536209, SD: ±66337105; MTEP mean: 156607143, SD: ±50820920; CHPG mean: 273645670, SD: ±84901986). H: Representative reconstruction of wild-type (N = 16) and grm5a-/- grm5b-/- (N = 17) expression of mbp:memGCaMP7s reporting dorsal (green) and Ventral axon myelination (yellow). I: Doral myelin volume in of wild-type and grm5a-/- grm5b-/- animals as volumetric pixel volume (vpx) (Unpaired t-test p-value = 0.0024). J: Relative Fluorescent intensity (RFU) in dorsal myelin sheaths of wild-type (N = 16, mean: 54180897, SD: ±29591313 and grm5a-/- grm5b-/- (N = 17, mean: 28549095, SD: ±13030576) animals expressing mbp:memGCaMP7s, reflecting baseline myelin Ca2+ activity (Unpaired t-test p-value = 0.0027). K: Ventral myelin volume in of wild-type (N = 16, mean: 944652 SD: ±252246) and grm5a-/- grm5b-/- (N = 17, mean: 470384, SD: ±219208) animals as volumetric pixel volume (vpx) (Unpaired t-test p-value = <0.0001). L: Relative Fluorescent intensity (RFU) in ventral myelin sheaths of wild-type (N = 16, mean: 246369587 SD: ±92539193) and grm5a-/- grm5b-/- (N = 17, mean: 88194157 SD: ±54784433 animals expressing mbp:memGCaMP7s, reflecting baseline myelin Ca2+ activity (Mann–Whitney test p-value = <0.0001).
