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

Journal: Frontiers in Oncology

Article Title: Repurposing riluzole as an anti-osteosarcoma agent

doi: 10.3389/fonc.2025.1508819

Figure Lengend Snippet: p53 expression and glutamate secretion in OS cells. (A) Western blots showing expression of p53 in OS cell lines and PDX cells. The data was analyzed by one-way ANOVA followed by Dunnett’s test. *p<0.05. (B–D) Glutamate secreted into media between days 0 and 7 by primary OS cell lines, hFOB and metastatic cell lines, and hFOB and PDX cells. The experiment was repeated three times; the average of the three is shown. The data was analyzed by one-way ANOVA followed by Dunnett’s test for each day, with the value for hFOB for that day serving as the comparison value. *p <0.05, **p<0.01, ***p<0.001. (E–F) mRNA expression of mGluR1 and mGluR5 by OS cell lines, measured by quantitative PCR. *p<0.05, **p<0.01, ***p<0.001. (G–H) Expression of vGLUT and xCT by OS cell lines and PDX cells, measured by western blot. The data was analyzed by one-way ANOVA followed by Dunnett’s post hoc analysis. ***p<0.001.

Article Snippet: For western blotting, we used antibodies for p53 (Proteintech #10442-1-AP), mGluR1 (Abclonal #A11462), mGluR5 (Santa Cruz Biotechnology #sc-293442), vGLUT1 (Abcam #ab272913), xCT (Abcam #ab307601), β-actin anti-mouse 8H10D10 (Cell Signaling Technologies #3700S), and GAPDH anti-rabbit (Cell Signaling Technology #5174S).

Techniques: Expressing, Western Blot, Comparison, Real-time Polymerase Chain Reaction

Journal: World Journal of Gastroenterology

Article Title: Inhibition of metabotropic glutamate receptor-5 alleviates hepatic steatosis by enhancing autophagy via activation of the AMPK signaling pathway

doi: 10.3748/wjg.v31.i7.98852

Figure Lengend Snippet: Inhibition of metabotropic glutamate receptor type 5 attenuates hepatocyte steatosis in vitro. A: Protein expression of metabotropic glutamate receptor type 5 (mGluR5) in HepG2 cells treated with different concentrations of free fatty acids (FFAs) for 24 hours; B: Protein expression of mGluR5 in HepG2 cells treated with 0.5 mmol/L FFAs for different Durations; C: CCK-8 assays revealed the effects of MPEP (2.5, 5, 10, 20, and 50 μM) on HepG2 cell viability; D: CCK-8 assays revealed the effects of CHPG (50, 100, 200, 500, and 1000 μM) on HepG2 cell viability; E and F: HepG2 cells were stimulated with 0.5 mmol/L FFAs for 24 hours and then treated with MPEP (10 μM) or CHPG (500 μM) for another 24 hours. The cells were stained with Oil Red O, and the intracellular triglyceride concentration was quantitatively determined. a P < 0.05 vs free fatty acids, b P < 0.01 vs free fatty acids, c P < 0.001 vs free fatty acids; d P < 0.001 vs bovine serum albumin. The data are presented as the mean ± SE. All cell experiments were repeated three times. mGluR5: Metabotropic glutamate receptor type 5; FFAs: Free fatty acids; BSA: Bovine serum albumin.

Article Snippet: As shown in Supplementary Figure 1C , to investigate the involvement of autophagy and AMPK in the inhibition of mGluR5, chloroquine (CQ) (HY-17589A, MedChemExpress) and compound C (CC) (HY-13418A, MedChemExpress) were separately added to the cells in the presence of FFAs for 24 hours after successful induction of steatosis.

Techniques: Inhibition, In Vitro, Expressing, CCK-8 Assay, Staining, Concentration Assay

Journal: World Journal of Gastroenterology

Article Title: Inhibition of metabotropic glutamate receptor-5 alleviates hepatic steatosis by enhancing autophagy via activation of the AMPK signaling pathway

doi: 10.3748/wjg.v31.i7.98852

Figure Lengend Snippet: Inhibition of metabotropic glutamate receptor type 5 improves lipid accumulation by increasing autophagy in free fatty acids-stimulated HepG2 cells. A and B: HepG2 cells were stimulated with 0.5 mmol/L free fatty acids (FFAs) for 24 hours and then treated with MPEP (10 μM) or CHPG (500 μM) for another 24 hours. Western blotting was used to detect the protein expression of LC3 and p62 in the cells; C and D: HepG2 cells were stimulated with 0.5 mmol/L FFAs for 24 hours and then treated with MPEP (10 μM) or MPEP (10 μM) +chloroquine (20 μM) for another 24 hours. The cells were stained with Oil Red O, and the intracellular triglyceride concentration was quantitatively determined. a P < 0.05 vs free fatty acids; b P < 0.01 vs free fatty acids; c P < 0.001 vs bovine serum albumin; d P < 0.001 vs MPEP; e P < 0.05 vs bovine serum albumin. The data are presented as the mean ± SE. All cell experiments were repeated three times. FFAs: Free fatty acids; BSA: Bovine serum albumin; CQ: Chloroquine.

Article Snippet: As shown in Supplementary Figure 1C , to investigate the involvement of autophagy and AMPK in the inhibition of mGluR5, chloroquine (CQ) (HY-17589A, MedChemExpress) and compound C (CC) (HY-13418A, MedChemExpress) were separately added to the cells in the presence of FFAs for 24 hours after successful induction of steatosis.

Techniques: Inhibition, Western Blot, Expressing, Staining, Concentration Assay

Journal: World Journal of Gastroenterology

Article Title: Inhibition of metabotropic glutamate receptor-5 alleviates hepatic steatosis by enhancing autophagy via activation of the AMPK signaling pathway

doi: 10.3748/wjg.v31.i7.98852

Figure Lengend Snippet: Inhibition of metabotropic glutamate receptor type 5-induced autophagy via activation of the AMPK signaling pathway. A and B: HepG2 cells were stimulated with 0.5 mmol/L free fatty acids (FFAs) for 24 hours and then treated with MPEP (10 μM) or CHPG (500 μM) for another 24 hours. Western blotting was used to detect the protein expression of P-AMPK, AMPK, P-ULK1 and ULK1 in the cells; C-E: HepG2 cells were stimulated with 0.5 mmol/L FFAs for 24 hours and then treated with MPEP (10 μM), CHPG or MPEP (10 μM) + Compound C (10 μM) for another 24 hours. The cells were stained with Oil Red O (C), and the level of intracellular triglyceride was quantitatively determined (D). Western blot was used to determine transcription factor EB expression in the cytoplasmic and nuclear fractions of HepG2 cells (E). Histone H3 was used as a reference control for nuclear expression. a P < 0.05 vs free fatty acids; b P < 0.01 vs free fatty acids; c P < 0.001 vs bovine serum albumin; d P < 0.01 vs bovine serum albumin; e P < 0.01 vs MPEP. The data are presented as the mean ± SE. All cell experiments were repeated three times. FFAs: Free fatty acids; BSA: Bovine serum albumin; TFEB: Transcription factor EB.

Article Snippet: As shown in Supplementary Figure 1C , to investigate the involvement of autophagy and AMPK in the inhibition of mGluR5, chloroquine (CQ) (HY-17589A, MedChemExpress) and compound C (CC) (HY-13418A, MedChemExpress) were separately added to the cells in the presence of FFAs for 24 hours after successful induction of steatosis.

Techniques: Inhibition, Activation Assay, Western Blot, Expressing, Staining, Control

Journal: World Journal of Gastroenterology

Article Title: Inhibition of metabotropic glutamate receptor-5 alleviates hepatic steatosis by enhancing autophagy via activation of the AMPK signaling pathway

doi: 10.3748/wjg.v31.i7.98852

Figure Lengend Snippet: Knockout of the metabotropic glutamate receptor type 5 gene improved glucose and lipid metabolism and induced hepatic autophagy in high-fat diet-fed mice. A: Weekly weights of male mice; B and C: Blood glucose curve and area under the curve of the Intraperitoneal glucose tolerance test in male mice; D: Serum insulin levels in male mice; E and F: Serum triglyceride and total cholesterol levels in male mice; G and H: Serum low-density lipoprotein and high-density lipoprotein levels in male mice; I: Western blot analysis was used to evaluate the protein expression levels of LC3 and p62 in mice livers. The data are presented as the mean ± SE. a P < 0.05 vs WT/NCD, b P < 0.01 vs WT/NCD, c P < 0.001 vs WT/NCD; d P < 0.05 vs WT/HFD, e P < 0.01 vs WT/HFD, f P < 0.001 vs WT/HFD. WT: Wild-type; KO: Knockout; HFD: High-fat diet; NCD: Normal chow diet; AUC: Area under the curve; TG: Triglyceride; TC: Total cholesterol; HDL-C: High-density lipoprotein cholesterol; LDL-C: Low-density lipoprotein cholesterol.

Article Snippet: As shown in Supplementary Figure 1C , to investigate the involvement of autophagy and AMPK in the inhibition of mGluR5, chloroquine (CQ) (HY-17589A, MedChemExpress) and compound C (CC) (HY-13418A, MedChemExpress) were separately added to the cells in the presence of FFAs for 24 hours after successful induction of steatosis.

Techniques: Knock-Out, Western Blot, Expressing

Journal: World Journal of Gastroenterology

Article Title: Inhibition of metabotropic glutamate receptor-5 alleviates hepatic steatosis by enhancing autophagy via activation of the AMPK signaling pathway

doi: 10.3748/wjg.v31.i7.98852

Figure Lengend Snippet: Schematic diagram summarizing the underlying mechanism by which metabotropic glutamate receptor type 5 alleviates hepatic steatosis. Metabotropic glutamate receptor type 5 inhibition promotes AMPK phosphorylation, which leads to increased LC3II expression and nuclear translocation of transcription factor EB and the induction of p62 transcription, thereby activating autophagy and reducing lipid accumulation in hepatocytes. mGluR5: Metabotropic glutamate receptor type 5; TFEB: Transcription factor EB.

Article Snippet: As shown in Supplementary Figure 1C , to investigate the involvement of autophagy and AMPK in the inhibition of mGluR5, chloroquine (CQ) (HY-17589A, MedChemExpress) and compound C (CC) (HY-13418A, MedChemExpress) were separately added to the cells in the presence of FFAs for 24 hours after successful induction of steatosis.

Techniques: Inhibition, Expressing, Translocation Assay