sv huc 1 cells  (Tocris)

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 , Protocol to label individual oligodendrocytes, manipulate mGluR5 activity and assess myelination. b , Relative frequency of sheath lengths (DMSO n = 427, mean 16.79 µm; MTEP n = 201, mean 18.41 µm; CHPG n = 470, mean 30.44 µm). c , Individual oligodendrocytes at 4 dpf, labeled with mbp:eGFP–CAAX, after treatment with DMSO, mGluR5 antagonist MTEP and allosteric agonist CHPG. Scale bar, 15 µm. d , Mean sheath length per oligodendrocyte (one OL/fish) post treatment (one-way analysis of variance (ANOVA), P < 0.0001; Holm–Šídák’s multiple comparisons test: DMSO versus CHPG P = 0.0062, DMSO versus MTEP P = 0.0047; CHPG versus MTEP P < 0.0001). 1% DMSO ( N = 23, 26.90 µm ± 6.031), CHPG ( N = 28, 31.66 µm ± 4.27) and MTEP ( N = 12, 20.15 µm ± 8.77). e , Number of myelin sheaths produced by single oligodendrocytes (one-way ANOVA; P = 0.2090, Kruskal–Wallis DMSO versus CHPG P = 0.2477; DMSO versus MTEP P > 0.9999; CHPG versus MTEP P > 0.9999). DMSO, N = 23, 18.04 ± 4.995; MTEP, N = 12, 16.75 ± 3.89; CHPG N = 28, 15.32 ± 6.08). Scale bar, 50 µm. f , Representative images of Tg(olig1:nls–mApple) after pharmacological manipulation of mGluR5. Scale bar, 50 µm. g , Number OPCs in the spinal cord (one-way ANOVA; P = 0.7774; Tukey’s multiple comparisons test, DMSO versus MTEP P = 0.9908; DMSO versus CHPG P = 0.8579; CHPG versus MTEP P = 0.7781), DMSO ( N = 15); CHPG ( N = 17); MTEP ( N = 16). DMSO 371.1 ± 62.39; MTEP 368.5 ± 54.37; CHPG 381.8 ± 53.13. h , Representative images of Tg(mbp:nls–eGFP) after pharmacological manipulation of mGluR5. i , Number of myelinating oligodendrocytes in the spinal cord (one-way ANOVA; P = 0.5155; Tukey’s multiple comparisons test: DMSO versus MTEP P = 0.9141; DMSO versus CHPG P = 0.7359; CHPG versus MTEP P = 0.5037), DMSO ( N = 22); CHPG ( N = 25); MTEP ( N = 18). DMSO, 252 ± 33.24; MTEP, 257.3 ± 43.61; CHPG, 243 ± 45.46. Scale bar, 50 µm. Data indicate 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: Activity Assay, Labeling, Produced

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

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 : Brightfield image of a 4 dpf old zebrafish. B Brightfield image of a 4 dpf grm5a −/− grm5b −/− mutant zebrafish. C : Swimming behavior of 5 dpf mGluR5 mutants and WT siblings in light and dark phases (WT N = 21 mean 863.8, SD: ±260.6; grm5a −/− N =14, mean: 860.3, SD: ±296.4; grm5b −/− N =13, mean: 856.7, SD: ±351.8; grm5a −/− grm5b −/− N =30, mean:860.3, SD: ±238.9). Average distances swam over time and distinct phases by different groups. D : Distance swam by individual fish during all dark phases combined (WT N = 21 mean 863.8, SD: ±260.6; grm5a -/- N=14, mean: 860.3, SD: ±296.4; grm5b -/- =13, mean: 856.7, SD: ±351.8; grm5a −/− grm5b -/- N=30, mean:860.3, SD: ±238.9). (One-Way ANOVA; p-value = 0.9999, Tukey’s multiple comparisons test: WT vs. grm5a -/- p-value = 0.9999, WT vs. grm5b -/- p-value = 0.9999, WT vs. grm5a -/- grm5b -/- p-value = 0.9999, grm5a -/- vs. grm5b -/- p-value = 0.9999, grm5a -/- vs. grm5a -/- grm5b -/- p-value = 0.9999,). E : Distance swam by individual fish during all light phases combined (WT N= 21 mean 348, SD: ±259.5; grm5a -/- N=14, mean: 313.9, SD: ±259.5; grm5b -/- N=13, mean: 390.8 SD: ±269.3; grm5a -/- grm5b -/- =30, mean:344.5, SD: ±253.3). (One-Way ANOVA; p-value = 0.2334; Tukey’s multiple comparisons test: WT vs. grm5a -/- p-value = 0.9807, WT vs. grm5b -/- p-value = 0.9650, WT vs. grm5a -/- grm5b -/- p-value = 0.9999 grm5a -/- vs. grm5b -/- p-value = 0.8653, grm5a -/- vs. grm5a -/- grm5b -/- p-value = >0.983, grm5a -/- vs. grm5a -/- grm5b -/- p-value = 0.9484).

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: Mutagenesis

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 : 3 and 4 dpf wild-type oligodendrocytes labeled with mpb:eGFP-CAAX (N = 36). B : 3 and 4 dpf grm5a -/- grm5b -/- oligodendrocytes at 3 dpf (N = 22). C : 3 and 4 dpf grm5a -/- oligodendrocytes (N = 14). D : 3 and 4 dpf grm5b -/- oligodendrocytes (N = 17). E: Relative frequency distribution of individual measured myelin sheath lengths in 3 dpf control and loss of function mutants (WT n = 583, mean: 20.03 µm) ( grm5a -/- n = 259, mean: 14.08 µm) ( grm5b -/- n = 279 mean: 13.48 µm) ( grm5a -/- grm5b -/- n = 338, mean: 15.89 µm). F: Relative frequency distribution of individual measured myelin sheath lengths in 4 dpf control and loss of function mutants (WT n = 288, mean: 27.52 µm; grm5a -/- n = 164, mean: 15.9 µm; grm5b -/- n = 209, mean: 19.72 µm; grm5a -/- grm5b -/- n = 191, mean: 18.88 µm). G Relative frequency distribution of individual measured myelin sheath length in 7 dpf WT (n:212 mean: 31.34 µm) and grm5a -/- grm5b -/- (n = 259, mean: 21.87) animals. H : Comparison between sheath lengths per cell in 3 and 4 dpf fish (One-Way ANOVA p-value > 0.0001; Tukey’s multiple comparisons test: 3 dpf WT vs. 3 dpf grm5a -/- p-value = 0.0038; 3 dpf WT vs. 3 dpf grm5b -/- p-value = 0.0064; WT 3 dpf vs. 3 dpf grm5a -/- grm5b -/- p-value = 0.0293; 3 dpf WT vs. 4 dpf WT: p-value = <0.0001; WT 4 dpf vs. 4 dpf grm5a -/- p-value = <0.0001; WT 4 dpf vs. 4 dpf grm5b -/- p-value = <0.0028; WT 4 dpf vs. 4 dpf grm5a -/- grm5b -/- p-value = 0.0002; 3 dpf WT vs 4 dpf grm5a -/- grm5b -/- p-value = 0.9842; 3 dpf grm5a -/- vs. 4 dpf WT p-value = <0.0001; 3 dpf grm5b -/- vs. 4 dps WT p-value = <0.0001, 3 dpf grm5a -/- grm5b -/- vs. 4 dpf WT p-value = <0.0001). WT 3 dpf mean: 21.11, SD: ±5.15; 3 dpf grm5a -/- mean= 14.5, SD: ±4.77; 3 dpf grm5b -/- mean= 15,17 SD: ±5.38; 3 dpf grm5a -/- grm5b -/- mean= 16.34 SD: ±5.376; 4 dpf WT mean: 28.8, SD: ±6.519; 4 dpf grm5a -/- mean: 15.99 SD: ±3.94, 4 dpf grm5b -/- mean: 20.83 SD: ±6.716; 4 dpf grm5a -/- grm5b -/- mean: 19.47, SD: ±3.67). I : Myelin sheath numbers of 3 and 4 dpf WT and mGluR5 mutants (One-way ANOVA p-value = 0.2457 Tukey’s multiple comparisons test: 3 dpf WT vs. 3 dpf grm5a -/- p-value = 0.8436; 3 dpf WT vs. 3 dpf grm5b -/- p-value = 0.9995; WT 3 dpf vs. 3 dpf grm5a -/- grm5b -/- p-value = 0.9988; 3 dpf WT vs. 4 dpf WT: p-value = >0.9999; WT 4 dpf vs. 4 dpf grm5a -/- p-value = 0.4523; WT 4 dpf vs. 4 dpf grm5b -/- p-value = 0.982; WT 4 dpf vs. 4 dpf grm5a -/- grm5b -/- p-value = >0.9999; 3 dpf WT vs. 4 dpf grm5a -/- grm5b -/- p-value = 0.9992, 3 dpf grm5a -/- vs. 4 dps WT p-value = 0.8797; 3 dpf grm5b -/- vs. 4 dps WT p-value = 0.9994, 3 dpf grm5a -/- grm5b -/- vs. 4 dpf WT p-value = 0.9999). WT 3 dpf mean: 16.2, SD: ±4.80; 3 dpf grm5a -/- mean: 18.5, SD: ±5.00 3 dpf grm5b -/- mean: 17, SD: ±4.74; 3 dpf grm5a -/- grm5b -/- mean: 15.36 SD: ±3.94; 4 dpf WT mean: 16.06 SD: ±4.04; 4 dpf grm5a -/- mean: 20.5 SD: ±8.23, 4 dpf grm5b -/- mean: 17.83 SD: ±7.49; 4 dpf grm5a -/- grm5b -/- mean: 15.92, SD: ±4.46).

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: Labeling, Control, Comparison

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 , Construct used to express mGluR5 tethered to eGFP, alongside a membrane anchored reporter mScarlet, in myelinating oligodendrocytes. b , Control construct used to express fluorescent reporter in myelinating oligodendrocytes. c , c ′, Images of oligodendrocytes expressing control mbp:memScarlet in wild-type (WT) ( c ) ( N = 15) and grm5a −/− ( c′ ) ( N = 20) animals at 4 dpf. d , d ′, Oligodendrocytes expressing mbp:memScarlet-P2A–grm5a–eGFP in WT ( d ) ( N = 12) and grm5a −/− ( d ′) ( N = 12) animals at 4 dpf. Scale bar, 10 μm. e , Relative frequency distribution of individual myelin sheath lengths. WT mbp:memScarlet n = 265 (mean 26.49 µm); WT mbp:memScarlet-P2A–grm5a–eGFP n = 124 (mean 31.55 µm); grm5a −/− mbp:memScarlet n = 365 (mean 19.54 µm); grm5a −/− mbp:memScarlet-P2A–grm5a–eGFP n = 138 (mean 31.58 µm). f , Mean sheath length of WT oligodendrocytes expressing mbp:memScarlet-P2A–grm5a–eGFP ( N = 12, 32.19 ± 8.41) or mbp:memScarlet ( N = 15, 26.92 ± 4.28) (two-sided unpaired t -test, P = 0.0445). g , Mean myelin sheath length of grm5a −/− oligodendrocytes expressing mbp:memScarlet-P2A–grm5a–eGFP ( N = 12, 33.25 ± 6.57) or mbp:memScarlet ( N = 20, 20.05 ± 3.45) (two-sided unpaired t -test, P = <0.0001). h , Sheath number per oligodendrocytes expressing mbp:memScarlet-P2A–grm5a–eGFP (12.67 ± 3.46) or mbp:memScarlet (17.13 ± 5.17) (two-sided unpaired t -test, P = 0.0164). i , Sheath number per oligodendrocytes in grm5a −/− mutants expressing mbp:memScarlet-P2A–grm5a–eGFP ( N = 12, 11.25 ± 4.33) or mbp:memScarlet ( N = 20, 17.1 ± 5.21) (two-sided unpaired t -test, P = 0.0027). Scale bar, 10 μm. 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: Construct, Membrane, Control, Expressing