Journal: Nature Neuroscience
Article Title: Activity-driven myelin sheath growth is mediated by mGluR5
doi: 10.1038/s41593-025-01956-9
Figure Lengend Snippet: a , HCR detection of grm5a and grm5b messenger RNA in OPCs at 5 dpf. Scale bar, 5 μm. b : HCR detection of grm5a and grm5b mRNA in OLs at 5 dpf. Scale bar, 5 μm. c , d , Schematic showing organization of human and zebrafish genes encoding mGluR5 and their CRISPR/Cas9 based editing that led to identification of mutants with premature STOP codons in exons 2 and 1 of grm5a ( c ) and grm5b ( d ), respectively. e , Images of wild-type ( N = 35) and grm5a −/− grm5b −/− ( N = 22) oligodendrocytes expressing mbp:eGFP–CAAX. f , Images of wild-type ( N = 18) and grm5a −/− grm5b −/− ( N = 12) oligodendrocytes expressing mbp:eGFP–CAAX at 4 dpf. g , Images of wild-type ( N = 10) and grm5a −/− grm5b −/− ( N = 14) oligodendrocytes expressing mbp:eGFP–CAAX at 7 dpf. Scale bars 15 μm ( e – g ). h , Mean sheath length of wild-type ( N = 35, 21.11 ± 5.149) and grm5a −/− grm5b −/− ( N = 22, 16.34 ± 5.38) oligodendrocytes, in 3 dpf old animals (two-sided Mann–Whitney U -test, P = 0.0035). i , Mean sheath length of wild-type ( N = 18, 28.8 ± 6.5) and grm5a −/− grm5b −/− ( N = 12, 19.47 ± 3.67) oligodendrocytes, in 4 dpf old animals (two-sided unpaired t -test: P = 0.0001). j , Mean sheath length of wild-type ( N = 10, 31.79 ± 4.723) and grm5a −/− grm5b −/− ( N = 14, 22.68 ± 4.40) oligodendrocytes, in 7 dpf old animals (two-sided unpaired t -test: P < 0.0001). k , Comparison of mean sheath length in wild-type animals between 3, 4 and 7 dpf animals (one-way ANOVA, Kruskal–Wallis test P < 0.0001; Dunn’s multiple comparisons test: wild-type 3 dpf versus 4 dpf P = 0.0002; wild-type 4 dpf to 7 dpf P = 0.7173, wild-type 3 dpf versus 7 dpf P < 0.0001). (3 dpf, 21.11 ± 5.149; 4 dpf, 28.8 ± 6.52; 7 dpf, 31.79 ± 4.72). l , Number of myelin sheath per oligodendrocyte in 3 dpf wild-type (16.2 ± 4.8) and grm5a −/− grm5b −/− (15.36 ± 3.94) (two-sided unpaired t -test; P = 0.4964). m , Number of sheaths per oligodendrocyte in 4 dpf wild-type (16.06 ± 4.03) and grm5a −/− grm5b −/− (15.92 ± 34.46) (two-sided unpaired t -test; P = 0.9301). n , Number of myelin sheath per oligodendrocyte in 7 dpf wild-type (21.2 ± 5.35) and grm5a −/− grm5b −/− (19.5 ± 8.91) (two-sided unpaired t -test; P = 0.5971). o , Comparison of the mean sheath length in grm5a −/− grm5b −/− animals between 3, 4 and 7 dpf animals (one-way ANOVA, Kruskal–Wallis test P < 0.0001; Dunn’s multiple comparisons test: grm5a −/− grm5b −/− 3 dpf versus 4 dpf P = 0.1675; grm5a −/− grm5b −/− 4 dpf to 7 dpf P = 0.2070, grm5a −/− grm5b −/− 3 dpf versus 7 dpf P < 0.0009) (3 dpf, 16.34 ± 5.38; 4 dpf, 19.47 ± 3.67; 7 dpf, 22.68 ± 4.40). Data show mean ± s.d.
Article Snippet: The following compounds were used in this study: the mGluR5 antagonist MTEP 3-((2-methyl-1,3-thiazol-4-yl)ethynyl)pyridine; Tocris 1186195-60-7; 200 μM, 1% DMSO vehicle), the mGluR5 agonist CHPG (RS)-2-chloro-5-hydroxyphenylglycine (Tocris Cas:170846-74-9; 360 μM, 1% DMSO vehicle), the sodium channel blocker MS222/Tricaine (Sigma; 600 μM, no vehicle), the neuromuscular blocking agent mivacurium chloride (Abcam; 1.5 mg ml −1 , no vehicle).
Techniques: CRISPR, Expressing, MANN-WHITNEY, Comparison