Journal: Heliyon

Article Title: The assessment of possible gender-related effect of endogenous striatal alpha-tocopherol level on MPTP neurotoxicity in mice

doi: 10.1016/j.heliyon.2020.e04425

Figure Lengend Snippet: DA, DOPAC and HVA concentration levels (ng/mg ww) and DA turnover in the striatum of mice.

Article Snippet: The reagents for αT high performance liquid chromatography (HPLC) measurement has already been reported [ ], and besides those, we used the following chemicals in this study: disodium-ethylenediaminetetraacetate dihydrate (Na EDTA∗2H O; Lach-Ner s.r.o, Neratovice, Czech Republic), sodium-metabisulfite (Na S O 5 ; Fluka, Budapest, Hungary, Honeywell Group), sodium-dihydrogenphosphate (NaH PO 4 ; Reanal Laboratory Chemicals, Budapest, Hungary), HPLC purity absolute ethanol (EtOH) and acetonitrile (ACN; VWR International, Radnar, PA, USA), MPTP hydrochloride (MedChemExpress, Monmouth Junction, NJ, USA) and the following substances were obtained from Sigma-Aldrich (Saint Louis, MO, USA): perchloric acid (HClO 4 ), phosphoric acid (H 3 PO 4 ), sodium hydroxide (NaOH), buthyl-hydroxy-toluol (BHT), sodium-octyl-sulphate (NaOS), 3,4-dihydroxyphenylacetic acid (DOPAC), 3,4-dihydroxybenzylamine hydrobromide (DHBA∗HBr, internal standard (IS)), homovanillic acid (HVA), dopamine hydrochloride (DA∗HCl) and isoproterenol hydrochloride (IPR∗HCl, IS).

Techniques: Concentration Assay, Control

Journal: Heliyon

Article Title: The assessment of possible gender-related effect of endogenous striatal alpha-tocopherol level on MPTP neurotoxicity in mice

doi: 10.1016/j.heliyon.2020.e04425

Figure Lengend Snippet: Measured analyte concentrations in mouse striatal and plasma samples. DA levels (a), DA turnover (b), αT levels in the striatum (c) and in the plasma (d) are presented for the demonstration of the effect of MPTP treatment. Data are presented as mean (±S.D.); n (control and MPTP-treated females) = 14; n (control males) = 15; n (MPTP-treated males) = 13; αT α-tocopherol; DA dopamine; DOPAC 3,4-dihydroxyphenylacetic acid; HVA homovanillic acid; MPTP 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; n.s. not significant; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: The reagents for αT high performance liquid chromatography (HPLC) measurement has already been reported [ ], and besides those, we used the following chemicals in this study: disodium-ethylenediaminetetraacetate dihydrate (Na EDTA∗2H O; Lach-Ner s.r.o, Neratovice, Czech Republic), sodium-metabisulfite (Na S O 5 ; Fluka, Budapest, Hungary, Honeywell Group), sodium-dihydrogenphosphate (NaH PO 4 ; Reanal Laboratory Chemicals, Budapest, Hungary), HPLC purity absolute ethanol (EtOH) and acetonitrile (ACN; VWR International, Radnar, PA, USA), MPTP hydrochloride (MedChemExpress, Monmouth Junction, NJ, USA) and the following substances were obtained from Sigma-Aldrich (Saint Louis, MO, USA): perchloric acid (HClO 4 ), phosphoric acid (H 3 PO 4 ), sodium hydroxide (NaOH), buthyl-hydroxy-toluol (BHT), sodium-octyl-sulphate (NaOS), 3,4-dihydroxyphenylacetic acid (DOPAC), 3,4-dihydroxybenzylamine hydrobromide (DHBA∗HBr, internal standard (IS)), homovanillic acid (HVA), dopamine hydrochloride (DA∗HCl) and isoproterenol hydrochloride (IPR∗HCl, IS).

Techniques: Clinical Proteomics, Control

Journal: Heliyon

Article Title: The assessment of possible gender-related effect of endogenous striatal alpha-tocopherol level on MPTP neurotoxicity in mice

doi: 10.1016/j.heliyon.2020.e04425

Figure Lengend Snippet: αT concentration levels in the plasma and the striatum of mice.

Article Snippet: The reagents for αT high performance liquid chromatography (HPLC) measurement has already been reported [ ], and besides those, we used the following chemicals in this study: disodium-ethylenediaminetetraacetate dihydrate (Na EDTA∗2H O; Lach-Ner s.r.o, Neratovice, Czech Republic), sodium-metabisulfite (Na S O 5 ; Fluka, Budapest, Hungary, Honeywell Group), sodium-dihydrogenphosphate (NaH PO 4 ; Reanal Laboratory Chemicals, Budapest, Hungary), HPLC purity absolute ethanol (EtOH) and acetonitrile (ACN; VWR International, Radnar, PA, USA), MPTP hydrochloride (MedChemExpress, Monmouth Junction, NJ, USA) and the following substances were obtained from Sigma-Aldrich (Saint Louis, MO, USA): perchloric acid (HClO 4 ), phosphoric acid (H 3 PO 4 ), sodium hydroxide (NaOH), buthyl-hydroxy-toluol (BHT), sodium-octyl-sulphate (NaOS), 3,4-dihydroxyphenylacetic acid (DOPAC), 3,4-dihydroxybenzylamine hydrobromide (DHBA∗HBr, internal standard (IS)), homovanillic acid (HVA), dopamine hydrochloride (DA∗HCl) and isoproterenol hydrochloride (IPR∗HCl, IS).

Techniques: Concentration Assay, Clinical Proteomics, Control

Journal: Microbiology Spectrum

Article Title: Neuroprotective effects of hypoactive Akkermansia muciniphila in MPTP-induced mouse models of Parkinson’s disease

doi: 10.1128/spectrum.03379-24

Figure Lengend Snippet: Investigation of the impact of hypoactive Akkermansia muciniphila on MPTP-induced behavioral alterations in mice. ( A ) Timeline of the experimental procedure; ( B ) longitudinal analysis of mouse body weight; ( C ) body weight gain following MPTP administration; ( D ) total descent duration in the pole test ( n = 7); ( E ) total walking duration in the beam walking test ( n = 7); ( F ) exploratory behavior in the Y-maze ( n = 7); ( G ) spontaneous alternation in the Y-maze ( n = 7). Comparative analysis of pre- and post-MPTP treatment performance in the pole test ( H ), beam walking test ( I ), and Y-maze ( J and K ). Data are presented as means ± SEM and were subjected to one-way ANOVA followed by Tukey’s post hoc test. * P < 0.05, indicating a significant difference compared to the saline group.

Article Snippet: Concurrently, the MPTP + AKK + L-DOPA group and MPTP + L-DOPA group received an additional oral gavage of L-DOPA (100 mg/kg; TargetMol) and benserazide (25 mg/kg; MedChemExpress).

Techniques: Saline

Journal: Microbiology Spectrum

Article Title: Neuroprotective effects of hypoactive Akkermansia muciniphila in MPTP-induced mouse models of Parkinson’s disease

doi: 10.1128/spectrum.03379-24

Figure Lengend Snippet: Investigation of the neuroprotective effects of hypoactive Akkermansia muciniphila and L-DOPA in the brain and colon. Panels A to F illustrate the mRNA expression levels of brain-derived neurotrophic factor (BDNF), Iba1, glial cell line-derived neurotrophic factor (GDNF), ZO-1, occludin, and claudin-1 in the brain ( n = 4). Panels G to J depict the mRNA expression levels of ZO-1, occludin, and claudin-1 in colon ( n = 4). Data are presented as means ± SEM and were analyzed using one-way ANOVA followed by Tukey’s post hoc test. Statistical significance is indicated by * P < 0.05 and ** P < 0.005.

Article Snippet: Concurrently, the MPTP + AKK + L-DOPA group and MPTP + L-DOPA group received an additional oral gavage of L-DOPA (100 mg/kg; TargetMol) and benserazide (25 mg/kg; MedChemExpress).

Techniques: Expressing, Derivative Assay

Journal: Microbiology Spectrum

Article Title: Neuroprotective effects of hypoactive Akkermansia muciniphila in MPTP-induced mouse models of Parkinson’s disease

doi: 10.1128/spectrum.03379-24

Figure Lengend Snippet: The effects of hypoactive Akkermansia muciniphila and L-DOPA on inflammation suppression. Panels A to D illustrate the mRNA expression levels of proinflammatory cytokines IL-6, TNF-a, IL-113, and the anti-inflammatory cytokine IL-10 in the brain ( n = 4). Panels E and F present the mRNA expression levels of IL-113 and IL-10 in the colon ( n = 4). Percentages of monocytes ( G ), lymphocytes ( H ), and neutrophilic granulocytes ( I ) are also depicted ( n = 6). Data are presented as means ± SEM and were analyzed using one-way ANOVA followed by Tukey’s post hoc test. Statistical significance is indicated as follows: * P < 0.05, ** P < 0.005, *** P < 0.0005, and **** P < 0.0001, in comparison to the MPTP group.

Article Snippet: Concurrently, the MPTP + AKK + L-DOPA group and MPTP + L-DOPA group received an additional oral gavage of L-DOPA (100 mg/kg; TargetMol) and benserazide (25 mg/kg; MedChemExpress).

Techniques: Expressing, Comparison

Journal: Microbiology Spectrum

Article Title: Neuroprotective effects of hypoactive Akkermansia muciniphila in MPTP-induced mouse models of Parkinson’s disease

doi: 10.1128/spectrum.03379-24

Figure Lengend Snippet: The fluctuations in the relative abundances of bacterial compositions in mouse fecal samples across various experimental time points. ( A ) A bar graph illustrates the relative compositions of the bacterial community at different time intervals ( n = 6). The relative abundances of fecal ( B ) Bacteroidota, ( C ) Firmicutes, ( D ) Proteobacteria, ( E ) Verrucomicrobiota, ( F ) Desulfobacterota, and ( G ) Akkermansia muciniphila are presented for each group throughout the experimental period ( n = 6). The data are presented as means ± SEM and were analyzed using two-way ANOVA followed by Tukey’s post hoc test and Dunn’s post hoc test. * P < 0.05, ** P < 0.005, *** P < 0.0005 denote significant differences ( n = 6). The blue/black asterisk indicates significant differences compared to the MPTP/saline group at each experimental time point; the gray asterisk indicates a significant difference within the same group between day 41 and day 34.

Article Snippet: Concurrently, the MPTP + AKK + L-DOPA group and MPTP + L-DOPA group received an additional oral gavage of L-DOPA (100 mg/kg; TargetMol) and benserazide (25 mg/kg; MedChemExpress).

Techniques: Saline

Journal: Microbiology Spectrum

Article Title: Neuroprotective effects of hypoactive Akkermansia muciniphila in MPTP-induced mouse models of Parkinson’s disease

doi: 10.1128/spectrum.03379-24

Figure Lengend Snippet: The administration of hypoactive Akkermansia muciniphila in conjunction with MPTP treatment significantly impacted the fecal microbiomes of mice. Variations in the diversity index among the groups were assessed using Chao1 ( A ) and ACE ( C ) estimator analyses ( n = 6). The data are expressed as means ± SEM and were evaluated using the Kruskal-Wallis test followed by Dunn’s post hoc test. A P -value of * P < 0.05 denotes a statistically significant difference within the same group between day 41 and day 34. Panel B presents a heat map illustrating the relative abundance of bacterial phyla in mice on day 41 ( n = 6). Panel D depicts the principal coordinate analysis of β-diversity, with axes labeled according to the contribution vectors (PCoA1 and PCoA2) of the principal components, and mouse samples are icon coded to distinguish between groups ( n = 3).

Article Snippet: Concurrently, the MPTP + AKK + L-DOPA group and MPTP + L-DOPA group received an additional oral gavage of L-DOPA (100 mg/kg; TargetMol) and benserazide (25 mg/kg; MedChemExpress).

Techniques: Labeling

Journal: Microbiology Spectrum

Article Title: Neuroprotective effects of hypoactive Akkermansia muciniphila in MPTP-induced mouse models of Parkinson’s disease

doi: 10.1128/spectrum.03379-24

Figure Lengend Snippet: The fluctuations in fecal short-chain fatty acid concentrations across all mouse groups throughout the experimental period. The concentrations, measured in micrograms of SCFA per gram of feces (gg/g), included ( A ) acetic acid, ( B ) propionic acid, ( C ) butyric acid, ( D ) isobutyric acid, ( E ) valeric acid, and ( F ) isovaleric acid. Data are presented as means ± SEM and were analyzed using a two-way ANOVA followed by Tukey’s post hoc test. Statistical significance is indicated as follows: * P < 0.05, ** P < 0.005, *** P < 0.0005 denote significant differences ( n = 6); the blue/black asterisk indicates significant differences compared to the MPTP/saline group at each experimental time point; the gray asterisk indicates a significant difference within the same group between day 41 and day 34.

Article Snippet: Concurrently, the MPTP + AKK + L-DOPA group and MPTP + L-DOPA group received an additional oral gavage of L-DOPA (100 mg/kg; TargetMol) and benserazide (25 mg/kg; MedChemExpress).

Techniques: Saline

Journal: Neural regeneration research

Article Title: CHCHD2 Thr61Ile mutation impairs F1F0-ATPase assembly in in vitro and in vivo models of Parkinson's disease.

doi: 10.4103/1673-5374.378010

Figure Lengend Snippet: Figure 2 ｜Cellular ATP levels and F1F0-ATPase activity in isolated mitochondria in a in SH-SY5Y cell model of Parkinson’s disease. (A, B) Flag-vector: LV-Flag-vector (Ubi-MCS-3FLAG-SV40-puromycin vector)-transfected SH-SY5Y cells cultured in MEM-F12 medium; Flag-vector + MPP+: LV-Flag-vector- transfected SH-SY5Y cells treated with MPP+ (500 μM for 24 hours); CHCHD2-Flag + MPP+: LV-CHCHD2-Flag-transfected SH-SY5Y cells treated with MPP+ (500 μM for 24 hours); CHCHD2-T61I + MPP+: LV-CHCHD2-T61I-Flag-transfected SH-SY5Y cells treated with MPP+ (500 μM for 24 hours). (A) Detection of ATP levels in control or MPP+-treated SH-SY5Y cells stably transfected with empty vector or constructs encoding WT or T61I-mutant CHCHD2. (B) Effect of WT or T61I-mutant CHCHD2 on ATP synthase specific activity in control or MPP+-treated cells. (C) Effect of WT or T61I-mutant CHCHD2 on ATP synthase specific activity in AVV-transfected mice with or without MPTP treatment. CHCHD2-Flag + MPTP: AVV-CHCHD2-Flag–transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); CHCHD2-T61I + MPTP: AVV-CHCHD2-T61I–transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); Flag-vector: AVV-Flag (CMV- betaGlobin-MCS-3Flag-SV40 polyA (GV411) vector)-transfected mice treated with normal saline (twice a week, for 5 weeks); Flag-vector + MPTP: AVV-Flag-transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks). Data are expressed as the mean ± SD (n = 3 independent experiments). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (one-way analysis of variance followed by Tukey’s post hoc test). ATP: Adenosine triphosphate; MPP+: 1-methyl-4-phenylpyridinium; MPTP: 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine.

Article Snippet: Detection of mitochondrial permeability transition pore opening Opening of the mitochondrial permeability transition pore (mPTP) was detected by the calcein-AM/cobalt assay following the manufacturer’s instructions (C2009S, Beyotime Biotechnology).

Techniques: Activity Assay, Isolation, Plasmid Preparation, Transfection, Cell Culture, Control, Stable Transfection, Construct, Mutagenesis, Saline

Journal: Neural regeneration research

Article Title: CHCHD2 Thr61Ile mutation impairs F1F0-ATPase assembly in in vitro and in vivo models of Parkinson's disease.

doi: 10.4103/1673-5374.378010

Figure Lengend Snippet: Figure 3 ｜CHCHD2 T61I mutation promotes mitochondrial permeability transition pore (mPTP) opening in SH-SY5Y cells. CHCHD2-Flag: LV-CHCHD2-Flag-transfected SH-SY5Y cells; CHCHD2-T61I: LV-CHCHD2-T61I-Flag-transfected SH-SY5Y cells; Flag-vector: LV-Flag-vector (Ubi-MCS-3FLAG-SV40-puromycin vector)-transfected SH-SY5Y cells. (A) Calcein-AM and CoCl2 were administered to live cells expressing WT or T61I-mutant CHCHD2, and fluorescence intensity was detected by flow cytometry. (B) Effect of WT or the T61I-mutant CHCHD2 on mPTP opening as determined by detecting mitochondrial membrane potential using JC-1 fluorescent probes (white squares) in control or MPP+-treated SH-SY5Y cells. The fluorescence intensity of intracellular JC-1 aggregates (red) in the MPP+ + CHCHD2-Flag group was greater than that in the MPP+ + Flag-vector group. The CHCHD2-T61I group showed an increased amount of JC-1 monomers (green). Scale bars: 10 μm. (C) The mean fluorescence intensity of JC-1 aggregates or monomers in control or MPP+-treated SH-SY5Y cells expressing WT or T61I-mutant CHCHD2. Data are expressed as the mean ± SD (n = 3 independent experiments). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (one-way analysis of variance followed by Tukey’s post hoc test). Calcein-AM: Calcein acetoxymethyl ester; MPP+: 1-methyl-4-phenylpyridinium; mPTP: mitochondrial permeability transition pore; WT: wild type.

Article Snippet: Detection of mitochondrial permeability transition pore opening Opening of the mitochondrial permeability transition pore (mPTP) was detected by the calcein-AM/cobalt assay following the manufacturer’s instructions (C2009S, Beyotime Biotechnology).

Techniques: Mutagenesis, Permeability, Transfection, Plasmid Preparation, Expressing, Fluorescence, Flow Cytometry, Membrane, Control

Journal: Neural regeneration research

Article Title: CHCHD2 Thr61Ile mutation impairs F1F0-ATPase assembly in in vitro and in vivo models of Parkinson's disease.

doi: 10.4103/1673-5374.378010

Figure Lengend Snippet: Figure 5 ｜CHCHD2 T61I mutation aggravates movement deficits and nigral DA neuron function in a mouse model of PD. (A) Experimental design. AAV-CHCHD2, AAV-CHCHD2 T61I, or AAV-Vector was stereotaxically injected into the substantia nigra pars compacta, and 4 weeks later mice were injected intraperitoneally with normal saline or MPTP for 5 weeks. One day after the last MPTP/normal saline injection, behavioral tests were performed, and then the mice were sacrificed. Flag-vector: AVV-Flag(CMV-betaGlobin-MCS-3Flag-SV40 polyA (GV411) vector)-transfected mice treated with normal saline (twice a week, for 5 weeks); Flag-vector+MPTP: AVV-Flag– transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); CHCHD2-Flag+MPTP: AVV-CHCHD2-Flag–infected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); CHCHD2-T61I+MPTP: AVV-CHCHD2-T61I–transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks). (B) The grasping test was used to examine grip strength. (C) The pole-climbing test was used to examine bradykinesia. (D) The rotarod test was used to examine motor coordination. (E) Representative immunohistochemical staining of TH-positive neurons in the substantia nigra (SN). There were fewer TH-positive neurons in the SN in the AAV-CHCHD2-T61I + MPTP group than in the AAV-Flag-vector + MPTP and AAV-CHCHD2-Flag + MPTP groups. Scale bars: 800 μm. (F) Quantification of the TH-positive cells shown in E. (G) Representative images of double-immunofluorescent staining for CHCHD2-Flag (green, Alexa Fluor 488) and TH (red, Alexa Fluor 555) (white squares) in the SN. There were significantly fewer TH-positive neurons in the SN in the AAV-CHCHD2-T61I + MPTP group than in the AAV-Flag-vector + MPTP and AAV-CHCHD2-Flag + MPTP groups. Scale bars: 50 μm. (H) Quantification of TH mean fluorescence in the SN as shown in G. (I) Representative immunoblot for TH in the SN. (J) TH expression in the SN. Results are expressed as the mean ± SD (n ≥ 6). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (one-way analysis of variance followed by Tukey’s post hoc test). ACTB: β-Actin; AAV: adeno-associated virus; DA: dopaminergic; DAPI: 4′,6-diamidino-2-phenylindole; MPTP: 1-methyl-4-phenyl- 1,2,3,6-tetrahydropyridine; TH: tyrosine hydroxylase; SN: substantia nigra.

Article Snippet: Detection of mitochondrial permeability transition pore opening Opening of the mitochondrial permeability transition pore (mPTP) was detected by the calcein-AM/cobalt assay following the manufacturer’s instructions (C2009S, Beyotime Biotechnology).

Techniques: Mutagenesis, Plasmid Preparation, Injection, Saline, Transfection, Infection, Immunohistochemical staining, Staining, Fluorescence, Western Blot, Expressing, Virus

Journal: Neural regeneration research

Article Title: CHCHD2 Thr61Ile mutation impairs F1F0-ATPase assembly in in vitro and in vivo models of Parkinson's disease.

doi: 10.4103/1673-5374.378010

Figure Lengend Snippet: Figure 8 ｜CHCHD2 modulates OSCP expression in MPP+-/MPTP-induced PD models. (A, B) Flag-vector: LV-Flag-vector (Ubi-MCS-3FLAG-SV40-puromycin vector)-transfected SH-SY5Y cells cultured in MEM-F12 medium; Flag-vector+MPP+: LV-Flag-vector–transfected SH- SY5Y cells treated with MPP+ (500 μM for 24 hours); CHCHD2-Flag+MPP+: LV-CHCHD2-Flag–transfected SH-SY5Y cells treated with MPP+ (500 μM for 24 hours); CHCHD2-T61I + MPP+: LV-CHCHD2-T61I-Flag–transfected SH-SY5Y cells treated with MPP+ (500 μM for 24 hours). (A) Representative immunoblot of OSCP, CHCHD2, and ACTB in MPP+-treated SH-SY5Y cells. (B) Quantification of relative OSCP expression shown in A. (C, D) Flag-vector: AVV-Flag(CMV-betaGlobin-MCS-3Flag-SV40 polyA (GV411) vector)-transfected mice treated with normal saline (twice a week, for 5 weeks); Flag-vector + MPTP: AVV-Flag-infected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); CHCHD2-Flag + MPTP: AVV-CHCHD2-Flag- infected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks); CHCHD2-T61I + MPTP: AVV-CHCHD2-T61I–transfected mice treated with MPTP (25 mg/kg, twice a week, for 5 weeks). (C) Representative immunoblot of OSCP, CHCHD2, and ACTB in control or MPTP-treated mice. (D) Quantification of relative OSCP expression shown in C. Data normalized to SH-SY5Y cells overexpressing Flag-vector or C57BL/6J mice transfected with AAV-Flag are expressed as the mean ± SD (n = 3 independent experiments for cells, n = 6 for animals). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (one-way analysis of variance followed by Tukey’s post hoc test). ACTB: β-Actin; MPP+: 1-methyl-4-phenylpyridinium; MPTP: 1-methyl- 4-phenyl-1,2,3,6-tetrahydropyridine; OSCP: oligomycin sensitivity conferral protein.

Article Snippet: Detection of mitochondrial permeability transition pore opening Opening of the mitochondrial permeability transition pore (mPTP) was detected by the calcein-AM/cobalt assay following the manufacturer’s instructions (C2009S, Beyotime Biotechnology).

Techniques: Expressing, Plasmid Preparation, Transfection, Cell Culture, Western Blot, Saline, Infection, Control

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: Experimental design. Three days before 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) injections, mice were pre-trained at the pole and beam tests by completing three trials within a day. Two days before MPTP injections, mice received an oral treatment with either vehicle [10% dimethyl sulfoxide (DMSO) and 25% Tween 20 v/v dissolved in saline 0.9% sodium chloride], FTY720 (1 mg/kg), or SEW2871 (20 mg/kg) once a day for a total of 14 days. At day 0, saline or MPTP (30 mg/kg/day) treatments were administered i.p. during 5 days. Motor abilities at the pole and beam tests were evaluated 8 days after the first MPTP injections. On day 11, all mice were sacrificed. The experimental setting involved six groups: 1) vehicle with saline treatments, n = 3; 2) FTY720 with saline treatments, n = 3; 3) SEW2871 with saline treatments, n = 3; 4) vehicle with MPTP treatments, n = 5; 5) FTY720 with MPTP treatments, n = 5; (6) SEW2871 with MPTP treatments, n = 5.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Saline

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: Levels of inflammatory markers are affected in the striatum. Levels of GFAP (A) , TNF-α (B) , and BDNF (C) were assessed by Western blot in the striatum of mice. The data, expressed relative to GAPDH, represent the mean of relative optical density in triplicate experiments of GFAP, TNF-α, and BDNF (expressed as a percentage of control values) ± S.E.M., n = 3–5 mice/group. ** p < 0.01, *** p < 0.001 vs . vehicle + saline; # p < 0.05, ## p < 0.01 vs . vehicle + MPTP.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Western Blot, Control, Saline

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: Signs of astrogliosis and microgliosis induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) are prevented by S1P1R modulators. (A) Representative examples of Iba-1 immunoreactive microglia (panel a1 to a4; red color), GFAP-positive astrocytes (panel a5 to a8; red color) and TH-positive neurons (panel a1 to a8; green color) using epifluorescence microscope (40X objective) in the mouse subtantia nigra pars reticulate (SNr; −3.16 mm from the Bregma). Bar equals 50 µm. Fluorescence intensity of Iba-1 and GFAP were measured in the SNc (B, D) as well as in the ventral tegmental area (VTA) (C, E) . The data represent the mean of Iba-1 and GFAP relative optical density (expressed as a percentage of control values) ± S.E.M., n = 3–5 mice/group. *** p < 0.001 vs . vehicle + saline; ### p < 0.001 vs . vehicle + MPTP.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Microscopy, Fluorescence, Control, Saline

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: S1PR modulators protect against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced subtantia nigra pars compacta (SNc) cellular loss. Representative examples of tyrosine hydroxylase (TH)-immunostaining (green) using the 40X objective of the epifluorescence microscope in coronal sections (−3.16 mm from the Bregma) of mouse subtantia nigra pars reticulate (SNr). Bar equals 50 µm (A) . Stereological counts of TH- and neuronal nuclear antigen (NeuN)-positive neurons in the SNc (B, D) and ventral tegmental area (VTA) (C, E) . The data represent the mean of TH- and NeuN-positive cell numbers/mm 3 (expressed as a percentage of control values) ± S.E.M., n = 3-5 mice/group. * p < 0.05, ** p < 0.01 vs . vehicle + saline; # p < 0.05, ## p < 0.01 vs . vehicle + MPTP.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Immunostaining, Microscopy, Control, Saline

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: Dopamine terminals were spared in the striatum following S1P1Rs modulators treatments. Levels of tyrosine hydroxylase (TH) (A) and dopamine transporter (DAT) (B) were determined in the mouse striatum by Western blot experiments. The data, expressed relative to GAPDH, represent the mean of relative optical density in triplicate experiments of TH and DAT (expressed as a percentage of control values) ± S.E.M., n = 3–5 mice/group. ** p < 0.01, *** p < 0.001 vs . vehicle + saline; ## p < 0.01 vs . vehicle + MPTP.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Western Blot, Control, Saline

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: Motor behavioral outcomes. Motor abilities on the pole and beam tests were evaluated. Data represent the mean time require to perform the pole (A) and beam (B) tests and the mean number of foot-faults occurring during the beam test (C) ± S.E.M., n = 3–5 mice/group. * p < 0.05, *** p < 0.001 vs . vehicle + saline; # p < 0.05, ### p < 0.001 vs . vehicle + MPTP.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Saline