FIGURE

Fig. 6

ID

ZDB-FIG-210217-30

Publication

Van Laar et al., 2020 - α-Synuclein amplifies cytoplasmic peroxide flux and oxidative stress provoked by mitochondrial inhibitors in CNS dopaminergic neurons in vivo

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Fig. 6

Fig. 6. TEMPOL prevents α-Synuclein-dependent cytoplasmic peroxide flux in dopaminergic neurons exposed to MPP⁺. A: Intravital confocal microscopy was used to image the roGFP2-Orp1 biosensor dynamically in the ventral diencephalon of live α-Syn; roGFP2-Orp1 zebrafish, following pretreatment with embryo water only (left six panels) or TEMPOL (3 mM in embryo water; right six panels), at baseline and following exposure to MPP⁺ (3 mM final bath concentration). Emission at 515 nm following excitation at 405 nm and 488 nm respectively are shown in the first two columns and the ratiometric 405/488 images (color scale to right of images) in the third column of each image set. B: Unbiased 3D spot detection was used to calculate the roGFP2-Orp1 405/488 ratio in dopaminergic DC4 – 6 neurons (n = 5 per group) in serial images acquired over 5 h after addition of MPP⁺ to the bath (x axis shows time after addition of MPP+). Mean ± SE roGFP2-Orp1 405/488 ratio is shown for neurons from α-Syn; Orp1-roGFP2 zebrafish that were pretreated with water only (blue) or TEMPOL (green). P < 0.05*, 0.01**, 0.0001**** water vs. TEMPOL at same time point, 2-way repeated measures ANOVA with Šidák multiple comparisons test. C: Ratiometric histograms showing the frequency distribution of the roGFP2-Orp1 405/488 ratio from every pixel within areas corresponding to DC4 – 6 dopaminergic neurons on 2D images at baseline (grey) and t = 300 min after addition of MPP⁺ for α-Syn zebrafish pretreated with water (blue) or TEMPOL (green). The arrowheads show the mean of each distribution. D: Scatterplot showing roGFP2-Orp1 405/488 ratio calculated using the 3D method in DC4 – 6 dopaminergic neurons from α-Syn zebrafish pre-treated with water (blue; n = 24) or TEMPOL (green; n = 31 neurons), at baseline and following exposure to MPP⁺. Data points represent individual neurons combined from 4 replicate zebrafish in each group; bars show mean ± SE; p < 0.0001****, 2-way ANOVA with Tukey multiple comparison test. E: Scatterplot showing fold increase in roGFP2-Orp1 405/488 ratio between baseline and post-MPP⁺ exposure for each individual cell shown in panel D. Bars show mean ± SE; p < 0.0001****, 2-tailed unpaired t-test.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	mitfa^w2/w2; mpv17^a9/a9; pt423Tg; zc57Tg
Conditions:	chemical treatment by environment: N-methyl-4-phenylpyridinium chemical treatment by environment: N-methyl-4-phenylpyridinium
Observed In:	dopaminergic neuron cell redox homeostasis
Stage:	Adult

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Redox Biol.