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Glucosylceramide-induced ectosomes propagate pathogenic α-synuclein in Parkinson’s disease

Nature Cell Biology volume 28, pages 492–506 (2026)Cite this article

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Abstract

The intercellular transmission of α-synuclein contributes to Parkinson’s disease pathology. Yet, the mechanisms of α-synuclein spread are not fully understood. Here we used live-cell microscopy to examine the impact of Parkinson’s disease associated lipid alterations on α-synuclein release. We discovered that increased glucosylceramides as a consequence of reduced β-glucocerebrosidase activity induce ectosome shedding from primary neurons and from dopaminergic neurons derived from induced pluripotent stem cells of a patient with Parkinson’s disease harbouring mutations in GBA1 (N370S, L444P and W378G) and LRRK2 (G2019S and R1441H) compared with their isogenic control. We show that elevated glucosylceramide and the pharmacological inhibition of β-glucocerebrosidase similarly increase vesicle release and uptake by other neurons in mouse brains. Finally, we show that ectosomes are loaded with α-synuclein and lead to the transmission of α-synuclein pathology to neighbouring neurons. These data reveal ectosomes as a major route for α-synuclein transmission in Parkinson’s disease.

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Fig. 1: Exogenous NBD–glucosylceramide or GCase inhibition induce ectosome formation.
Fig. 2: Glucosylceramide-induced ectosome formation in an in vivo mouse brain.
Fig. 3: Dopaminergic neurons derived from PD-patient iPS cells shed more ectosomes.
Fig. 4: Increasing GCase activity or blocking glucosylceramide synthesis suppresses ectosome formation from PD neurons.
Fig. 5: Improving lysosomal functions through reacidification or LRRK2 kinase inhibition prevents ectosome formation.
Fig. 6: Ectosomes from cortical neurons accumulate α-synuclein fibrils.
Fig. 7: Ectosomes shed by PD neurons contain pathogenic α-synuclein.
Fig. 8: Ectosomes shed by PD neurons transmit α-synuclein pathology.

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Data availability

Data supporting this work are available in the Article. Uncropped blots (Source Data Fig. 1), raw data values (Source Data Table 1), statistics information (Supplemental Table 3) and light microscopy reporting table (Supplementary Table 4) are provided with the Article. All other data supporting the findings of this study, including imaging files, are available from the corresponding author upon request. Source data are provided with this paper.

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Acknowledgements

We thank P. McPherson, H. Bellen, G. Lin, X. Pan, M. Gu, M. Tyrlik, A. Walimbe, S. Kaduskar and S.-A. Mok for helpful comments on the manuscript. We thank X. Sun, P. Gao and S. van Baarle for assistance with electron microscopy and A. Simmonds for the use of Imaris software. We thank J. Han and his team of the Uvic-Genome BC Proteomics Centre for the sphingolipid analysis and K. Jacquemyn for the support related to serialEM software. Experiments were performed at the Faculty of Medicine and Dentistry Cell Imaging Core RRID: SCR_019200. We thank E. Tayler and L. Kravitz for vector art distributed on SciDraw (https://doi.org/10.5281/zenodo.3925901). This work was supported by the Canadian Institutes of Health Research (grant nos. 173321 and 191990), the Canada Research Chairs Programme (grant no. 2021-00027) and a Future Leaders in Canadian Brain Research from Brain Canada (grant no. 5946). T.D. was supported by GBA1 Canada (G-CAN) research and tools and development funding. J. Jacquemyn was supported by a Dr. Rowland and Muriel Haryett Neuroscience Fellowship and an EMBO Postdoctoral Fellowship (grant no. ALTF 120-2022).

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Authors and Affiliations

  1. Department of Physiology, University of Alberta, Edmonton, Alberta, Canada

    Julie Jacquemyn, Brian Marriott, Jinlan Chang, Ehlam Iftikhar, Kennedi Chik, Nathanael Y. J. Lee, Luis F. Rubio Atonal, Ceili Green, Jeremy Wong, Claudia Acevedo-Morantes, Jesse Jackson & Maria S. Ioannou

  2. Department of Cell Biology, University of Alberta, Edmonton, Alberta, Canada

    Julie Jacquemyn & Maria S. Ioannou

  3. Group on Molecular and Cell Biology of Lipids, University of Alberta, Edmonton, Alberta, Canada

    Julie Jacquemyn, Nathanael Y. J. Lee & Maria S. Ioannou

  4. Neuroscience and Mental Health Institute, University of Alberta, Edmonton, Alberta, Canada

    Julie Jacquemyn, Brian Marriott, Nathanael Y. J. Lee, Jesse Jackson & Maria S. Ioannou

  5. The Neuro’s Early Drug Discovery Unit, McGill University, Montreal, Quebec, Canada

    Carol X.-Q. Chen, Michael Nicouleau, Zhipeng You, Eric Deneault, Narges Abdian & Thomas M. Durcan

  6. Centre for Oncology, Radiopharmaceuticals and Research, Biologic and Radiopharmaceutical Drugs Directorate, Health Products and Food Branch, Health Canada, Ottawa, Ontario, Canada

    Eric Deneault

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Contributions

Conceptualization: M.S.I., J. Jacquemyn. Methodology: M.S.I., J. Jacquemyn, B.M., J. Jackson, C.X.Q.C., T.D. Investigation: J. Jackson. Technical assistance: J.C., E.I., K.C., N.Y.L., L.F.R.A., C.G., J.W., C.A.M. Resources: N.A., M.N., E.D., Z.Y. Visualization: M.S.I., J. Jacquemyn. Funding acquisition: M.S.I., J. Jacquemyn, T.D. Supervision: M.S.I., J. Jackson, T.D. Writing—original and revised draft: M.S.I., J. Jacquemyn. Writing—review and editing: M.S.I., J. Jacquemyn, J. Jackson, C.X.Q.C., T.D.

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Correspondence to Maria S. Ioannou.

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Nature Cell Biology thanks Dan Li and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Extended data

Extended Data Fig. 1 Increased glucosylceramide and reduced GCase activity induces ectosome formation in neurons.

a, Live-cell image of cortical neuron expressing mCh-GPI treated with NBD-glucosylceramide (NBD-GlcCer). Boxed area highlighting vesicles budding from soma and magnified on the left. b, Intensity of ethidium homodimer-1 (EthD-1) staining of neurons treated with NBD-GlcCer relative to control. n = 4 independent experiments; mean ± SEM; Two-tailed one sample t-test. c, Cytotoxicity assessment of NBD-GlcCer on cortical neurons by MTT assay. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; Two-tailed Mann-Whitney test. d, Time-lapse imaging showing mCh-GPI positive ectosome biogenesis with NBD-GlcCer treatment. e, Images of attached and detached ectosomes containing intraluminal vesicles labeled with mCh-GPI and NBD-GlcCer from the neuron-conditioned media. f, GCase activity of cortical neurons treated with CBE. The plotted line is a non-linear fit of values. n = 4 independent experiments. g, Heatmap showing fold change of significantly altered HexCer (galactosylceramides and glucosylceramides) of neuron treated with 100 µM CBE. n = 5 independent experiments. h, Average fold change of HexCer species with CBE treatment. n = 5 independent experiments. P values calculated using unpaired two-tailed t-test with false discovery rate adjusted using Benjamini-Hockberg method. i, Live-cell image of cortical neuron expressing mVenus-CAAX treated with 3 µM CBE, to induce 50% activity reduction. The arrows highlight vesicles budding from the plasma membrane along the neurites. Boxes show magnified images on the right. j, Amount of mVenus-CAAX ectosome buds per cell area relative to control. n = 4 independent experiments; 10 cells/coverslip/treatment; mean ± SEM; Two-tailed one sample t-test. k-m, Size distribution of attached and detached mCh-GPI and mVenus-CAAX-positive vesicles from neurons treated with 3 and 100 µM CBE. n = 4 independent experiments; for 3 and 100 µM CBE a total of 111 attached, 231 attached and 64 detached were analyzed respectively; mean ± SEM. n-o, Neurons were treated with CBE and stained with ethidium homodimer-1 (EthD-1) and assessed for cytotoxicity by MTT assay. n = 4 independent experiments; mean ± SEM; One-way ANOVA with Dunn’s multiple comparisons test. p, GCase activity of cortical astrocytes treated with CBE. The plotted line is a non-linear fit of values. n = 4 independent experiments; mean ± SEM. q, Live-cell image of cortical astrocytes expressing mVenus-CAAX treated with 100 µM CBE. Boxes show magnified images with orthogonal views on the right confirming ectosome budding from plasma membrane. r, Amount of mVenus-CAAX ectosome buds per astrocyte area relative to control. n = 4 independent experiments; 10 cells/coverslip/treatment; mean ± SEM; Two-tailed one sample t-test. s, Representative image of detached mVenus-CAAX positive ectosome from the astrocyte-conditioned media. 4 independent experiments were performed. Graphs are depicted as superplots where biological replicates are shown in large shapes, and technical replicates are shown as small shapes.

Source data

Extended Data Fig. 2 Ectosomes containing tdTomato fluorophore are released by cortical neurons treated with glucosylceramide in vivo.

a, Percentage of NBD-GlcCer positive buds containing the tdTomato fluorophore. n = 4 independent experiments; mean ± SEM. b, Coronal section of mouse brain with saline or NBD-GlcCer injection. Tiled widefield and fluorescent images are displayed. Boxes indicate the saline or NBD-GlcCer injection site. c, Maximum intensity projection of fixed tissue showing a cortical neuron expressing tdTomato after saline administration. Boxed area shows magnification of tdTomato positive vesicle, ≥ 2 µM from neuron, with orthogonal views on the right confirming it is fully detached. d, Number of tdTomato positive puncta in NeuN positive cells. n = 3 animals; 5 images per n; mean ± SEM; Two-tailed unpaired t-test. e, Number of NBD-GlcCer positive puncta in NeuN positive cells. n = 3 animals; 5 images per n; mean ± SEM; Two-tailed unpaired t-test. f, Percentage of tdTomato positive puncta with NBD-GlcCer in Neun positive cells neighboring the tdTomato positive cortical neuron. n = 3 animals; 5 images per n; mean ± SEM; Two-tailed unpaired t-test. Graphs are depicted as superplots where biological replicates are shown in large shapes, and where applicable technical replicates are shown as small shapes.

Extended Data Fig. 3 Increased mVenus-CAAX-positive ectosomes from GBA1 and LRRK2 patient-derived dopaminergic neurons.

a, iPS cell-derived dopaminergic neurons fixed and immunostained for β3-tubulin and tyrosine hydroxylase. b, Percentage of tyrosine hydroxylase positive dopaminergic neurons in GBA1 and LRRK2 mutant lines and their isogenic controls. n = 4 independent inductions; mean ± SEM. c, Live-cell maximum intensity projection of LRRK2-G2019S and isogenic corrected control (corr) iPS cell-derived dopaminergic neurons expressing mVenus-CAAX. Boxed areas highlighting ectosomes forming at the plasma membrane are magnified below. d, Number of mVenus-CAAX positive buds per cell area in LRRK2-G2019S and control iPS cell-derived dopaminergic neuron. n = 3 independent experiments; mean ± SEM; Two-tailed one sample t-test. e-f, Size distribution of attached mVenus-CAAX-positive vesicles in GBA1 and LRRK2 mutant lines. independent experiments: n = 3 for GBA1 (total 118 vesicles) and n = 4 for LRRK2 (total 239 vesicles); mean ± SEM. g-h Cytotoxicity assessment of mVenus-CAAX expression in GBA1 and LRRK2 mutant lines and their isogenic controls with MTT assay. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; One-way ANOVA with Dunn’s multiple comparison. Graphs are depicted as superplots where biological replicates are shown in large shapes, and where applicable technical replicates are shown as small shapes.

Extended Data Fig. 4 Increased tubulin-positive ectosomes from GBA1 and LRRK2 patient-derived dopaminergic neurons.

a, Schematic of differential ultracentrifugation procedure used to separate extracellular vesicles (EVs). b-d, Western blotting of centrifugation fractions prepared from the GBA1-N370S mutant and its isogenic control as in A. CL, cell lysate; S, supernatant; P, pellet. Quantification of CD81 and tubulin present in 10,000 x g fractionated pellet normalized to the cell lysate and relative to the isogenic control. n = 3 independent experiments; mean ± SEM; Two-tailed one sample t-test. Graph shows the experimental averages. e, Percentage mVenus-CAAX positive buds containing tubulin induced by CBE. n = 3 independent experiments; mean ± SEM; Two-tailed unpaired t-test. f, Live-cell image of cortical neurons expressing mVenus-CAAX treated with SiR-tubulin and CBE. Boxed areas highlighting an ectosome containing tubulin is magnified below. g, Size distribution of mVenus-CAAX-positive ectosome buds from neurons treated with SiR-tubulin and 100 µM CBE. n = 4 independent experiments and a total of 77 attached vesicles were analyzed. h, Live-cell image of the GBA1-N370S neuron expressing mVenus-CAAX treated with SiR-tubulin. Boxed areas highlighting an ectosome containing tubulin is magnified below. i, Percentage mVenus-CAAX positive buds containing tubulin in GBA1-N370S mutant and its isogenic corrected control. n = 3 independent experiments; mean ± SEM; Two-tailed unpaired t-test. A total of 113 attached vesicles were analyzed. j, Size distribution of mVenus-CAAX-positive ectosome buds in SiR-tubulin treated GBA1-N370S neurons. k-m, Cytotoxicity assessment of SiR-tubulin treatment on mVenus-CAAX expressing GBA1-N370S and control neurons, as well as in NBD-glucosylceramide (NBD-GlcCer) or CBE treated cortical neurons by MTT assay. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; One-way ANOVA with Dunn’s multiple comparison and Two-tailed Mann-Whitney test. n-q, Maximum intensity projection of GBA1-N370S iPS cell-derived dopaminergic neurons with Tubb3 and caveolin-1 or clathrin. Boxed areas showing single plane magnified tubulin-positive buds containing clathrin and caveolin-1. Percentage tubulin positive buds containing clathrin and caveolin-1 in GBA1-N370S neuron. n = 4 independent experiments; mean ± SEM. r, Maximum intensity projections of GBA1-N370S dopaminergic neurons and isogenic controls +/- Pitstop 2 treatment, stained for β3-tubulin. Arrows indicate ectosomes budding from the neurite and soma. s, Quantification of tubulin-positive buds relative to control with Pitstop 2 treatment. n = 4 independent experiments; 8 cells per experiment; Mean ± SEM; One-way ANOVA with Šidák’s correction for multiple comparisons. Graphs are depicted as superplots where biological replicates are shown in large shapes, and technical replicates are shown as small shapes.

Extended Data Fig. 5 Validation of hGCase/eGFP expression in dopaminergic neurons.

a-b, Cytotoxicity assessment of the GCase modulator 758 on GBA1-N370S, LRRK2-G2019S and corresponding isogenic control neurons by MTT. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; One-way ANOVA with Dunn’s multiple comparison. c-h, Western blotting of GBA1-N370S, LRRK2-G2019S and corresponding isogenic control neurons expressing hGCase/eGFP or eGFP and immunoblotted for hGCase, GFP and β-actin. Expression of hGCase and eGFP normalized against β-actin. n = 4 independent experiments, mean ± SEM. i-j, Percentage of eGFP positive dopaminergic neurons in GBA1-N370S, LRRK2-G2019S and corresponding isogenic controls. n = 4 independent experiments; 4 tile scans /2 coverslip/experiment; mean ± SEM. k-l, Cytotoxicity assessment of hGCase/eGFP expression on GBA1-N370S, LRRK2-G2019S and corresponding isogenic control neurons by MTT. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; One-way ANOVA with Dunn’s multiple comparison. Graphs are depicted as superplots where biological replicates are shown in large shapes, and where applicable technical replicates are shown as small shapes.

Extended Data Fig. 6 Sphingolipid profiling of lysates and large vesicle fraction containing ectosomes in dopaminergic neurons.

a, Schematic overview of sphingolipid pathway, species are color coded, GCase and important lipids in bold. b-q, Abundance of lipids in lysates and large extracellular vesicle fractions (lEV) from GBA1-N370S neurons and isogenic corrected controls. n = 5 independent experiments from separate inductions; Mean ± SEM; Two-tailed unpaired t-tests with Šidák’s correction for multiple comparisons. Graphs are depicted as superplots where biological replicates are shown in large shapes, and technical replicates are shown as small shapes.

Extended Data Fig. 7 Ectosome formation is inhibited by ibiglustat and MLi-2, but unaffected by glucosylsphingosine or carmofur.

a, Live-cell images of cortical neurons expressing mVenus-CAAX +/- glucosylsphingosine (GlcSph). Magnified images of neurites on the right. b, Amount of mVenus-CAAX ectosome buds per cell area relative to control with GlcSph treatment. n = 4 independent experiments; 10 cells/coverslip/treatment; mean ± SEM; Two-tailed one sample t-test. c, Cytotoxicity assessment of GlcSph on cortical neurons by MTT assay. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; Two-tailed Mann-Whitney test. d, Quantification of average diameter of attached tubulin-positive vesicles in GBA1-N370S +/- carmofur. n = 4 independent experiments; mean ± SEM; One-way ANOVA with Dunn’s multiple comparison. e, Cytotoxicity assessment of carmofur (Car) treatment on dopaminergic neurons by MTT assay. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; One-way ANOVA with Šidák’s correction for multiple comparisons. f-g, Quantification of average diameter of attached tubulin-positive vesicles in GBA1-N370S +/- ibiglustat. n = 4 independent experiments; mean ± SEM; One-way ANOVA with Dunn’s multiple comparison. h-i, Cytotoxicity assessment of ibiglustat on GBA1-N370S, LRRK2-G2019S and corresponding isogenic control neurons by MTT. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; One-way ANOVA with Dunn’s multiple comparison. j-k, Cytotoxicity assessment of rimeporide on GBA1-N370S, LRRK2-G2019S and corresponding isogenic control neurons by MTT. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; One-way ANOVA with Dunn’s multiple comparison. l, Cytotoxicity assessment of LRRK2 kinase inhibitor, MLi-2, on LRRK2-G2019S and corresponding isogenic control neurons by MTT. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; One-way ANOVA with Dunn’s multiple comparison. Graphs are depicted as superplots where biological replicates are shown in large shapes, and where applicable technical replicates are shown as small shapes.

Extended Data Fig. 8 Validation of α-synuclein pre-formed fibrils.

a-b, Thioflavin T increases emission at 482 nm with pre-formed fibrils (PFFs) generated from monomeric α-synuclein indicating the presence of beta sheet-rich structures which are absent from monomeric or PBS controls. 2x, double concentration. n = 6-8 independent experiments; mean ± SEM. c, Transmission electron micrographs of α-synuclein PFFs before and after sonication. d-e, Length distribution of fluorescently labeled and unlabeled PFFs after sonication. f, Percent area of pS129-α-synuclein relative to β3-tubulin in primary cortical neurons over time. n = 4 independent experiments; mean ± SEM; two-way ANOVA with Dunn’s multiple comparison test. g-h, Images of cortical cultures at DIV14 treated with PBS, monomeric α-synuclein, tagged PFFs (Atto-594) or untagged PFFs stained for β3-tubulin and pS129-α-synuclein. Boxed areas are magnified below and on the right. i, Representative live-cell image of ATTO594-α-synuclein fibrils in ectosomes formed upon addition of NBD-GlcCer in cortical neurons. Boxed areas are magnified on the left. 4 independent experiments were performed.

Extended Data Fig. 9 Ectosomes as a vehicle for the transmission of pathogenic α-.

a-b, Cytotoxicity assessment of PFFs on NBD-GlcCer and CBE treated cortical neurons expressing mVenus-CAAX by MTT assay. n = 4 independent experiments; 3 technical replicates/experiment; mean ± SEM; Two-tailed Mann-Whitney test and One-way ANOVA with Dunn’s multiple comparison. c, Number of mVenus-CAAX positive vesicles per area in GBA1 isogenic control neurons +/-PFFs. Independent experiments: n = 3 control and n = 4 for PFF-ATTO594 treatment; 10 cells/coverslip/treatment; mean ± SEM; Two-tailed one sample t-test. d, Cytotoxicity assessment of PFFs on mVenus-CAAX expressing GBA1-N370S and isogenic control neurons by MTT. n = 6 independent experiments; 3 technical replicates/experiment; mean ± SEM; One-way ANOVA with Dunn’s multiple comparison. e-f, Western blotting of centrifugation fractions derived from GBA1-N370S and control neurons treated with PFFs and immunoblotted for flotilin-1, pS129-α-synuclein and synuclein. CL, cell lysate; S, Supernatant; P, Pellet. 2 K, 2000 x g; 10 K, 10,000 x g; 100k, 100,000 x g. n = 3 independent experiments. g, Procedures used to separate vesicles and low molecular weight α-synuclein released by GBA1-N370S neurons by differential centrifugation. h, Image of fixed GBA1-N370S dopaminergic showing pS129-α-synuclein in tubulin-positive ectosomal bud. Boxed areas are magnified on the right. i, Percentage of tubulin-positive buds containing pS129-α-synuclein in PFFs60-treated GBA1-N370S dopaminergic. n = 4 independent experiments; 10 cells/experiment. Mean ± SEM. Graphs are depicted as superplots where biological replicates are shown in large shapes, and technical replicates are shown as small shapes.

Extended Data Fig. 10 Ectosome-mediated propagation of phosphorylated α-synuclein in GBA1-N370S neurons.

a-b, Maximum intensity projections of GBA1-N370S and control neurons incubated with large vesicle fraction derived from GBA1-N370S + /- PFF for 14 days and stained with β3-tubulin and pS129-α-synuclein. Quantifications of the amount of intracellular pS129-α-synuclein normalized to the neuron area in GBA1-N370S and control neurons treated with different centrifugation fractions after 14 days of incubation. n = 3 independent experiments; 6 cells/experiment. Mean ± SEM, One-way ANOVA with uncorrected Fisher’s LSD. c, Schematic representation of the multi-step propagation of pathogenic α-synuclein through large vesicles/ectosomes. d-e, Quantifications of the amount of intracellular pS129-α-synuclein normalized to the neuron area in GBA1-N370S and control neurons treated with large vesicle fraction from multi-step process depicted in (C) after 7 days of incubation. n = 4 independent experiments; 8 cells/experiment. Mean ± SEM; One-way ANOVA with uncorrected Fisher’s LSD. Max intensity projections of GBA1-N370S and control neurons incubated with the second-generation large vesicle fractions derived from GBA1-N370S. f-h, Size distribution of attached β3-tubulin-positive vesicles from GBA1-N370S + /- GW4869 and corresponding isogenic control neurons. n = 4 independent experiments; a total of 138, 199 and 234 attached vesicles for isogenic control and GBA1-N370S + /- GW4869 were analyzed; bars show mean ± SEM. i, Average size of vesicle diameter of GBA1-N370S + /- GW4869. n = 4 independent experiments; bars show mean ± SEM; Two-tailed Mann-Whitney test. j, Cytotoxicity assessment of GW4869 on GBA1-N370S and isogenic control neurons by MTT assay. n = 4 independent experiments; 3 technical replicates per experiment; bars show mean ± SEM; One-way ANOVA, with Dunn’s multiple correction test. k, Percentage of Iba1 positive microglia neighboring the tdTomato-expressing neuron containing tdTomato puncta. n = 5 animals; 6 images per n; mean ± SEM; Two-tailed unpaired t-test. l, Maximum intensity projection showing a cortical neuron expressing tdTomato after saline or CBE administration and immunostained for Iba1 and DAPI. Boxed area, maximum intensity projection, single slice magnification and orthogonal views (YZ; scale bar 2 µm) showing tdTomato-positive puncta internalized by neighboring microglia stained with Iba1. Graphs are depicted as superplots where biological replicates are shown in large shapes, and where applicable technical replicates are shown as small shapes.

Supplementary information

Supplementary Information (download PDF )

Supplementary Figs. 1–10. List and availability information for all iPS cell lines used in the study. Validation and characterization of new lines generated.

Reporting Summary (download PDF )

Supplementary Video (download MP4 )

Supplementary Video 1. A super-resolution microscopy image of a coronal brain section showing detached tdTomato-NBD-GlcCer positive puncta in three dimensions.

Supplementary Table (download XLSX )

Supplementary Table 1. List of significantly altered HexCer species, as determined by lipidomics, in CBE-treated cortical neurons versus control neurons.

Supplementary Table (download XLSX )

Supplementary Table 2. List of sphingolipid species in ectosome fraction and cell lysates of GBA1-N370S dopaminergic neurons and corresponding isogenic control.

Supplementary Table (download XLSX )

Supplementary Table 3. Statistics used for the study.

Supplementary Table (download XLSX )

Supplementary Table 4. Light microscopy reporting table.

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Source Data Fig. 1 uncropped blots. Unprocessed western blots.

Source Data Extended Data Table 1 (download XLSX )

Source Data Table 1 source data. Raw data.

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Jacquemyn, J., Marriott, B., Chang, J. et al. Glucosylceramide-induced ectosomes propagate pathogenic α-synuclein in Parkinson’s disease. Nat Cell Biol 28, 492–506 (2026). https://doi.org/10.1038/s41556-026-01871-6

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