Journal: Neural Regeneration Research

Article Title: Comparative proteomic analysis of plasma exosomes reveals the functional contribution of N-acetyl-alpha-glucosaminidase to Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01500

Figure Lengend Snippet: Primers for quantitative reverse transcription-polymerase chain reaction

Article Snippet: The sections were then incubated with antibodies to tyrosine hydroxylase (TH) (rabbit, 1:200, Abcam, Cat# ab137869, RRID: AB_2801410), NAGLU (rabbit, 1:500, Abcam, Cat# ab214671), and α-syn (mouse, 1:200, Santa Cruz Biotechnology, Cat# sc-53955, RRID: AB_831743).

Techniques:

Journal: Neural Regeneration Research

Article Title: Comparative proteomic analysis of plasma exosomes reveals the functional contribution of N-acetyl-alpha-glucosaminidase to Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01500

Figure Lengend Snippet: NAGLU and α-syn expression in MPP + -treated N2A cells and α-syn-overexpressing SH-SY5Y cells. (A) Immunoblot analysis of NAGLU and α-syn expression in untreated and MPP + -treated N2A cells. β-actin was used as a loading control. (B, C) Relative protein (B) and mRNA expression (C) of NAGLU and α-syn. (D) Immunoblot analysis of NAGLU and α-syn expression in untransfected cells (control), cells transfected with a control vector (vector), and cells transfected with plasmids encoding α-syn WT (WT) or α-syn A53T (A53T). β-Actin was used as a loading control. (E) Relative expression of NAGLU and α-syn. (G) Double-immunostaining for NAGLU (green) and α-syn (red) in untransfected SH-SY5Y cells (control), SH-SY5Y cells transfected with a control vector (vector), and SH-SY5Y cells transfected with plasmids encoding α-syn WT (WT) or α-syn A53T (A53T). Nuclei were stained with DAPI (blue). Scale bars: 50 μm. (H) NAGLU and α-syn fluorescence intensity. * P < 0.05, ** P < 0.01, *** P < 0.001 (unpaired t -test for B, C, and H; one-way analysis of variance followed by Tukey’s post hoc test for E, F). Data are shown as the mean ± SEM ( n = 3). DAPI: 4′,6-Diamidino-2-phenylindole; MPP + : methyl-4-phenylpyridinium; NAGLU: N-acetyl-alpha-glucosaminidase; N.S.: not significant; WB: western blot; WT: wild-type; α-syn: α-synuclein.

Article Snippet: The sections were then incubated with antibodies to tyrosine hydroxylase (TH) (rabbit, 1:200, Abcam, Cat# ab137869, RRID: AB_2801410), NAGLU (rabbit, 1:500, Abcam, Cat# ab214671), and α-syn (mouse, 1:200, Santa Cruz Biotechnology, Cat# sc-53955, RRID: AB_831743).

Techniques: Expressing, Western Blot, Control, Transfection, Plasmid Preparation, Double Immunostaining, Staining, Fluorescence

Journal: Neural Regeneration Research

Article Title: Comparative proteomic analysis of plasma exosomes reveals the functional contribution of N-acetyl-alpha-glucosaminidase to Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01500

Figure Lengend Snippet: NAGLU overexpression protects N2A cells against the effects of MPP + in vitro. (A) Western blot showing that NAGLU expression was significantly elevated in cells transfected with an NAGLU overexpression plasmid compared with untransfected cells and cells transfected with a control vector. β-Actin was used as a loading control. (B) Relative NAGLU expression. (C) Relative NAGLU mRNA expression normalized to the control. (D) Cells were transfected with an NAGLU overexpression plasmid or a control vector and treated with MPP + (500 μM) for 24 hours, and cell survival was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay. (E) NAGLU and α-syn expression in each group. β-Actin was used as a loading control. (F, G) NAGLU and α-syn expression. (H) Double-immunostaining for NAGLU (green) and α-syn (red) in N2A cells. Nuclei (blue) were counterstained with DAPI. Scale bar: 50 μm. (I) NAGLU and α-syn fluorescence intensities are shown. Data are presented as the mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s post hoc test). DAPI: 4′,6-Diamidino-2-phenylindole; MPP + : methyl-4-phenylpyridinium; NAGLU: N-acetyl-alpha-glucosaminidase; N.S.: not significant; OE: overexpression; α-syn: α-synuclein.

Article Snippet: The sections were then incubated with antibodies to tyrosine hydroxylase (TH) (rabbit, 1:200, Abcam, Cat# ab137869, RRID: AB_2801410), NAGLU (rabbit, 1:500, Abcam, Cat# ab214671), and α-syn (mouse, 1:200, Santa Cruz Biotechnology, Cat# sc-53955, RRID: AB_831743).

Techniques: Over Expression, In Vitro, Western Blot, Expressing, Transfection, Plasmid Preparation, Control, MTT Assay, Double Immunostaining, Fluorescence

Journal: Neural Regeneration Research

Article Title: Comparative proteomic analysis of plasma exosomes reveals the functional contribution of N-acetyl-alpha-glucosaminidase to Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01500

Figure Lengend Snippet: NAGLU expression was reduced in the brains of MPTP-treated mice. (A) Representative immunofluorescence staining images of the substantia nigra pars compacta showing that the number of TH+ cells (red) was decreased in MPTP-treated mice. Nuclei (blue) were counterstained with DAPI. (B, D) TH, NAGLU, and α-syn fluorescence intensities. (C, E) α-syn (red) (C) and NAGLU (red) (E) immunofluorescence in the midbrain. Scale bar: 200 μm. (F) Relative NAGLU and α-syn mRNA expression (qRT-PCR). Results are presented as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01 (unpaired t -test). DAPI: 4′,6-Diamidino-2-phenylindole; TH: tyrosine hydroxylase; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; NAGLU: N-acetyl-alpha-glucosaminidase; α-syn: α-synuclein.

Article Snippet: The sections were then incubated with antibodies to tyrosine hydroxylase (TH) (rabbit, 1:200, Abcam, Cat# ab137869, RRID: AB_2801410), NAGLU (rabbit, 1:500, Abcam, Cat# ab214671), and α-syn (mouse, 1:200, Santa Cruz Biotechnology, Cat# sc-53955, RRID: AB_831743).

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Quantitative RT-PCR

Journal: Neural Regeneration Research

Article Title: Comparative proteomic analysis of plasma exosomes reveals the functional contribution of N-acetyl-alpha-glucosaminidase to Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01500

Figure Lengend Snippet: Evaluation of the diagnostic value of exosomal NAGLU in PD. (A) Plasma exosome NAGLU levels were significantly lower in patients with PD ( n = 63) than in healthy controls ( n = 66), as determined by ELISA (Student’s t -test). (B) Receiver operating characteristic curves for exosome NAGLU expression levels between patients with PD and healthy controls. (C) Exosomal NAGLU levels at different H–Y stages (one-way analysis of variance followed by Tukey’s post hoc test). (D) Correlation analysis of H–Y stages and NAGLU level. Correlations between exosome α-syn and NAGLU expression in patients with PD (E) and controls (F). * P < 0.05, ** P < 0.01, *** P < 0.001. Pearson correlation analysis was used in D, E, and F. H–Y: Hoehn and Yahr stage; NAGLU: N-acetyl-alpha-glucosaminidase; N.S.: not significant; PD: Parkinson’s disease.

Article Snippet: The sections were then incubated with antibodies to tyrosine hydroxylase (TH) (rabbit, 1:200, Abcam, Cat# ab137869, RRID: AB_2801410), NAGLU (rabbit, 1:500, Abcam, Cat# ab214671), and α-syn (mouse, 1:200, Santa Cruz Biotechnology, Cat# sc-53955, RRID: AB_831743).

Techniques: Diagnostic Assay, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Expressing