Fig. 6: Mitochondrial dysfunction due to prolonged mitochondria–lysosome contacts is partially rescued by expression of TBC1D15 in GBA1-PD patient dopaminergic neurons. | Nature Communications

Fig. 6: Mitochondrial dysfunction due to prolonged mitochondria–lysosome contacts is partially rescued by expression of TBC1D15 in GBA1-PD patient dopaminergic neurons.

From: Dysregulation of mitochondria-lysosome contacts by GBA1 dysfunction in dopaminergic neuronal models of Parkinson’s disease

Fig. 6

a Live-cell distribution of mitochondria (TMRM, red) in the soma and axons from PD patient-derived mutant GBA1 dopaminergic neurons (∆GBA) and its CRISPR-corrected isogenic control (Corr) neurons. White arrows mark mitochondria in a single axon. Scale bar, 10 μm. b Quantified mitochondrial density in the soma (TMRM-positive pixels/pixels of soma). Corr (n = 47 neurons), ∆GBA (n = 47 neurons) (N = 3 independent experiments). c Quantified mitochondrial density in axons (mitochondria count/length of axon (μm)). Corr (n = 122 axons), ∆GBA (n = 114 axons) (N = 3 independent experiments). b, c Unpaired two-sided Student’s t-test; ***p < 0.0001. d Oxygen consumption rate (OCR) was measured and normalized to total protein content. Corr (n = 9 samples), ∆GBA (n = 9 samples) (N = 3 independent experiments). e Western blot analysis of phospho-AMPKα and AMPKα levels in Corr and ∆GBA neurons. Ratio of p-AMPKα/AMPKα are expressed as fold-change compared to Corr (N = 3 independent experiments). Paired two-sided Student’s t-test; *p = 0.0354. f Total cellular ATP content was measured and normalized to total protein content (ng/μl). Corr (n = 40 samples), ∆GBA (n = 40 samples) (N = 3 independent experiments). Unpaired two-sided Student’s t-test; ***p < 0.0001. g Live-cell distribution of mitochondria (TMRM, red) in the soma and axons from Corr and ∆GBA human neurons with or without human TBC1D15 lentiviral expression. White arrows mark mitochondria in a single axon. Scale bar, 10 μm. h Quantification of average minimum duration of M–L contacts in Corr-vehicle (n = 62 contacts from 18 neurons), ∆GBA-vehicle (n = 60 contacts from 15 neurons), ∆GBA-TBC1D15 (n = 63 contacts from 19 neurons) (N = 3 independent experiments); ***p < 0.0001. i Quantified mitochondrial density in the soma (TMRM-positive pixels/pixels of soma). Corr-vehicle (n = 32 neurons), ∆GBA-vehicle (n = 31 neurons), ∆GBA-TBC1D15 (n = 38 neurons) (N = 3 independent experiments). j Quantified mitochondrial density in axons (mitochondria count/length of axon (μm)). Corr-vehicle (n = 124 axons), ∆GBA-vehicle (n = 127 axons), ∆GBA-TBC1D15 (n = 127 axons) (N = 3 independent experiments), ***p < 0.0001. k Total cellular ATP concentration was measured and normalized to total protein content (ng/μl). Corr-vehicle (n = 19 samples), ∆GBA-vehicle (n = 19 samples), ∆GBA-TBC1D15 (n = 19 samples) (N = 3 independent experiments), ***p = 0.0002, *p = 0.0493. h–k One-way ANOVA followed by Tukey’s multiple comparisons test. For all quantifications, data are the means ± S.E.M.; *p ≤ 0.05, ***p ≤ 0.001, ns: not significant.

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