CoChat
Sign Up
Cited by 100

The DOI matches its Crossref record; 22 of the 22 references it cites were found in a registry or an index.

Parkinson’s Disease–Associated LRRK2 Interferes with Astrocyte-Mediated Alpha-Synuclein Clearance

Streubel-Gallasch, Linn, Giusti, Veronica, Sandre, Michele, Tessari, Isabella, Plotegher, Nicoletta, Giusto, Elena, Masato, Anna, Iovino, Ludovica, Battisti, Ilaria, Arrigoni, Giorgio, Shimshek, Derya, Greggio, Elisa, Tremblay, Marie-Eve, Bubacco, Luigi, Erlandsson, Anna, Civiero, Laura·2021·Molecular Neurobiology
DOI 10.1007/s12035-021-02327-8PMC PMC8257537

Published version of a preprint

Open this paper in CoChat — free account

Abstract

Parkinson’s disease (PD) is a neurodegenerative, progressive disease without a cure. To prevent PD onset or at least limit neurodegeneration, a better understanding of the underlying cellular and molecular disease mechanisms is crucial. Mutations in the leucine-rich repeat kinase 2 ( LRRK2 ) gene represent one of the most common causes of familial PD. In addition, LRRK2 variants are risk factors for sporadic PD, making LRRK2 an attractive therapeutic target. Mutations in LRRK2 have been linked to impaired alpha-synuclein (α-syn) degradation in neurons. However, in which way pathogenic LRRK2 affects α-syn clearance by astrocytes, the major glial cell type of the brain, remains unclear. The impact of astrocytes on PD progression has received more attention and recent data indicate that astrocytes play a key role in α-syn-mediated pathology. In the present study, we aimed to compare the capacity of wild-type astrocytes and astrocytes carrying the PD-linked G2019S mutation in Lrrk2 to ingest and degrade fibrillary α-syn. For this purpose, we used two different astrocyte culture systems that were exposed to sonicated α-syn for 24 h and analyzed directly after the α-syn pulse or 6 days later. To elucidate the impact of LRRK2 on α-syn clearance, we performed various analyses, including complementary imaging, transmission electron microscopy, and proteomic approaches. Our results show that astrocytes carrying the G2019S mutation in Lrrk2 exhibit a decreased capacity to internalize and degrade fibrillar α-syn via the endo-lysosomal pathway. In addition, we demonstrate that the reduction of α-syn internalization in the Lrrk2 G2019S astrocytes is linked to annexin A2 (AnxA2) loss of function. Together, our findings reveal that astrocytic LRRK2 contributes to the clearance of extracellular α-syn aggregates through an AnxA2-dependent mechanism.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1007/s12035-021-02327-8

Cites

22 of 22 references verified0 citation errors · 0 not checkable

Statistics

Figures

Fig. 1 Accumulation of aggregated α-syn in Lrrk2 astrocytes. A Schematic outline of the experimental setup. Cells were exposed to 0.5 μM sonicated Cy3 α-syn PFFs for 24 h. B TEM images of α-syn PFFs pre and post sonication. Scale bars 1 μm. C Representative fluorescence microscopy images of Lrrk1+/+, Lrrk2−/−, and Lrrk2GS/GS astrocytes (GFAP, green) at 24 h; cell nuclei stained with DAPI (blue) and α-syn labeled with Cy3 (red). Insets show a close-up of Cy3 α-syn inclusions. D Orthogonal projections of z-stack images taken with a fluorescence microscope: main view (x/y), top (x/z), and right (y/z). Projections were made along the lines depicted in the main image. Astrocytes (GFAP, green), Cy3 labeled α-syn (red), DAPI (blue). E Fluorescence microscopy image showing the differently sized Cy3 α-syn inclusions observed: small dot-like inclusions (arrow head) and larger, cottony deposits (arrow). E’ Displays of the particle count obtained from the ImageJ analysis when including either all Cy3 α-syn deposits or only the small Cy3 α-syn inclusions. Scale bars = 20 μm (C, D, and E)
Fig. 1 Accumulation of aggregated α-syn in Lrrk2 astrocytes. A Schematic outline of the experimental setup. Cells were exposed to 0.5 μM sonicated Cy3 α-syn PFFs for 24 h. B TEM images of α-syn PFFs pre and post sonication. Scale bars 1 μm. C Representative fluorescence microscopy images of Lrrk1+/+, Lrrk2−/−, and Lrrk2GS/GS astrocytes (GFAP, green) at 24 h; cell nuclei stained with DAPI (blue) and α-syn labeled with Cy3 (red). Insets show a close-up of Cy3 α-syn inclusions. D Orthogonal projections of z-stack images taken with a fluorescence microscope: main view (x/y), top (x/z), and right (y/z). Projections were made along the lines depicted in the main image. Astrocytes (GFAP, green), Cy3 labeled α-syn (red), DAPI (blue). E Fluorescence microscopy image showing the differently sized Cy3 α-syn inclusions observed: small dot-like inclusions (arrow head) and larger, cottony deposits (arrow). E’ Displays of the particle count obtained from the ImageJ analysis when including either all Cy3 α-syn deposits or only the small Cy3 α-syn inclusions. Scale bars = 20 μm (C, D, and E)CC BY · PubMed Central
Fig. 2 Analysis of the differently sized α-syn inclusions in Lrrk2 astrocytes. A, B Analysis of small and large Cy3 α-syn inclusions, respectively. Quantifications of Cy3 α-syn particle count, total area, and integrated density were performed using ImageJ. For both time points, ten images per independent cell culture (24 h: Lrrk1+/+, n = 7; Lrrk2−/−, n = 6; and Lrrk2GS/GS, n = 5; 24 h + 6 d: Lrrk1+/+, n = 6; Lrrk2−/−, n = 6; and Lrrk2GS/GS, n = 5) were analyzed and reported. For each time point, the statistical analysis was performed with the Kruskal-Wallis test followed by Dunn’s multiple comparisons test, since the data did not follow Gaussian distribution for all groups. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001
Fig. 2 Analysis of the differently sized α-syn inclusions in Lrrk2 astrocytes. A, B Analysis of small and large Cy3 α-syn inclusions, respectively. Quantifications of Cy3 α-syn particle count, total area, and integrated density were performed using ImageJ. For both time points, ten images per independent cell culture (24 h: Lrrk1+/+, n = 7; Lrrk2−/−, n = 6; and Lrrk2GS/GS, n = 5; 24 h + 6 d: Lrrk1+/+, n = 6; Lrrk2−/−, n = 6; and Lrrk2GS/GS, n = 5) were analyzed and reported. For each time point, the statistical analysis was performed with the Kruskal-Wallis test followed by Dunn’s multiple comparisons test, since the data did not follow Gaussian distribution for all groups. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001CC BY · PubMed Central
Fig. 3 Internalization of aggregated α-syn in striatal astrocytes. A TEM images of SNARF-1 α-syn PFFs pre and post sonication. B Schematic outline of the experimental setup. Cells were exposed to 0.5 μM sonicated SNARF-1 α-syn PFFs for 24 h and imaged using live confocal laser scanning microscopy. C Representative images of primary Lrrk2+/+ striatal astrocytes treated with SNARF-1 α-syn PFFs were acquired at range of 530–550 and 610–630 nm. Scale bar 50 μm. D, E Eight images per cell culture were analyzed (n = 4 independent cultures). For each image, ROIs were traced and quantification of α-syn single-particle 550/630 ratio (IntDen) and number were performed using ImageJ. The cumulative distribution of the single-particle ratio was graphed for each genotype in D and particle number per ROI in E. Statistical analysis in E was performed using Kruskal-Wallis test followed by Dunn’s multiple comparisons test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001
Fig. 3 Internalization of aggregated α-syn in striatal astrocytes. A TEM images of SNARF-1 α-syn PFFs pre and post sonication. B Schematic outline of the experimental setup. Cells were exposed to 0.5 μM sonicated SNARF-1 α-syn PFFs for 24 h and imaged using live confocal laser scanning microscopy. C Representative images of primary Lrrk2+/+ striatal astrocytes treated with SNARF-1 α-syn PFFs were acquired at range of 530–550 and 610–630 nm. Scale bar 50 μm. D, E Eight images per cell culture were analyzed (n = 4 independent cultures). For each image, ROIs were traced and quantification of α-syn single-particle 550/630 ratio (IntDen) and number were performed using ImageJ. The cumulative distribution of the single-particle ratio was graphed for each genotype in D and particle number per ROI in E. Statistical analysis in E was performed using Kruskal-Wallis test followed by Dunn’s multiple comparisons test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001CC BY · PubMed Central
Fig. 4 Description of the endo-lysosomal pathway in Lrrk2 striatal astrocytes. A Representative TEM image of primary striatal astrocyte section containing electron-dense lysosomal-like structures (arrows). Scale bar 2 μm. B, C Forty TEM images were acquired (n = 4 per genotype). Each cell was imaged by covering the entire cytoplasm and lysosomal-like structure number and area were measured using ImageJ. D Representative image of the staining using Lamp2A (green) as a marker for the endo-lysosomal pathway, DAPI (blue) for the nuclei, and F-actin (cyano) to define cells. Scale bar 20 μm. Inset shows a close-up of Lamp2A-positive structures. Quantifications of Lamp2A-positive structure number and area were analyzed using ImageJ. E, F Four images per cell culture were analyzed (n = 3 per genotype). G Measurement of lysosomal pH was done in primary striatal astrocytes from the three genotypes upon unlabeled α-syn PPF treatment (n = 3 per genotype). Bafilomycin has been applied as negative control. Fluorescence ratio of light acquired at 535 nm upon excitation at 340 and 380 nm is provided. H Neutral red assay was performed in primary striatal astrocytes from the three genotypes upon unlabeled α-syn PPF treatment (n = 4 per genotype). Bafilomycin has been applied as negative control. Absorbance at 540 nm measured upon cell lysates was normalized by total protein content. Statistical analysis in B, C, and F was made by Kruskal-Wallis test followed by Dunn’s multiple comparisons test. Statistical analysis in E, G, and H was performed with one-way ANOVA followed by Tukey’s multiple comparisons test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001
Fig. 4 Description of the endo-lysosomal pathway in Lrrk2 striatal astrocytes. A Representative TEM image of primary striatal astrocyte section containing electron-dense lysosomal-like structures (arrows). Scale bar 2 μm. B, C Forty TEM images were acquired (n = 4 per genotype). Each cell was imaged by covering the entire cytoplasm and lysosomal-like structure number and area were measured using ImageJ. D Representative image of the staining using Lamp2A (green) as a marker for the endo-lysosomal pathway, DAPI (blue) for the nuclei, and F-actin (cyano) to define cells. Scale bar 20 μm. Inset shows a close-up of Lamp2A-positive structures. Quantifications of Lamp2A-positive structure number and area were analyzed using ImageJ. E, F Four images per cell culture were analyzed (n = 3 per genotype). G Measurement of lysosomal pH was done in primary striatal astrocytes from the three genotypes upon unlabeled α-syn PPF treatment (n = 3 per genotype). Bafilomycin has been applied as negative control. Fluorescence ratio of light acquired at 535 nm upon excitation at 340 and 380 nm is provided. H Neutral red assay was performed in primary striatal astrocytes from the three genotypes upon unlabeled α-syn PPF treatment (n = 4 per genotype). Bafilomycin has been applied as negative control. Absorbance at 540 nm measured upon cell lysates was normalized by total protein content. Statistical analysis in B, C, and F was made by Kruskal-Wallis test followed by Dunn’s multiple comparisons test. Statistical analysis in E, G, and H was performed with one-way ANOVA followed by Tukey’s multiple comparisons test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001CC BY · PubMed Central
Fig. 5 Characterization of LRRK2 interactoma in stimulated condition. A Schematic outline of the experimental setup. H4 cells were transfected using 3xFlag-LRRK2 encoding plasmid and, after 48 h post transfection, treated with 0.5 μM sonicated unlabeled α-syn PFFs for 24 h. LRRK2 was subsequently immunopurified using anti-Flag agarose beads (IP), eluted with Flag peptide (elution), and subjected to LC-MS/MS analysis. FT, flow-through; unbound LRRK2. B Western blot analysis showing LRRK2 expression, immunopurification, and elution in H4 cells in treated and basal conditions. C Relative quantification of LRRK2 interactome under treated and untreated conditions. The area of the precursor ions identified by LC-MS/MS analysis was used as a quantitative measure of the protein content. The ratio between the area of the precursor ions of untreated and treated samples (normalized by the content of LRRK2) was then considered to highlight proteins showing a different affinity for LRRK2 in the two conditions (n = 2). D H4 cells transfected with GFP-ANXA2 in α-syn PFF-treated condition verifying the proximity of internalized α-syn fibrils and transfected AnxA2. AnxA2-GFP (green), α-syn (red), DAPI (blue). Scale bar 30 μm
Fig. 5 Characterization of LRRK2 interactoma in stimulated condition. A Schematic outline of the experimental setup. H4 cells were transfected using 3xFlag-LRRK2 encoding plasmid and, after 48 h post transfection, treated with 0.5 μM sonicated unlabeled α-syn PFFs for 24 h. LRRK2 was subsequently immunopurified using anti-Flag agarose beads (IP), eluted with Flag peptide (elution), and subjected to LC-MS/MS analysis. FT, flow-through; unbound LRRK2. B Western blot analysis showing LRRK2 expression, immunopurification, and elution in H4 cells in treated and basal conditions. C Relative quantification of LRRK2 interactome under treated and untreated conditions. The area of the precursor ions identified by LC-MS/MS analysis was used as a quantitative measure of the protein content. The ratio between the area of the precursor ions of untreated and treated samples (normalized by the content of LRRK2) was then considered to highlight proteins showing a different affinity for LRRK2 in the two conditions (n = 2). D H4 cells transfected with GFP-ANXA2 in α-syn PFF-treated condition verifying the proximity of internalized α-syn fibrils and transfected AnxA2. AnxA2-GFP (green), α-syn (red), DAPI (blue). Scale bar 30 μmCC BY · PubMed Central
Fig. 6 Investigation of AnxA2 function in astrocyte-mediated α-syn phagocytic clearance. A Unlabeled α-syn PFFs have been applied to primary astrocytes for 24 h. Projections verify the proximity of the internalized α-syn fibrils and endogenous AnxA2. Cell cytoskeleton (F-actin, cyano), α-syn (red), AnxA2 (green), DAPI (blue). Scale bar 20 μm. B Schematic outline of the experimental setup. Cells were transfected using 3xFlag-GFP-encoding plasmid together with scramble or AnxA2 siRNA. 48 h post transfection cells were treated with 0.5 μM sonicated α-syn PFFs for 24 h and imaged using confocal microscopy. C Representative images of primary striatal astrocytes transfected with scramble or Anxa2 siRNA together with a GFP-encoding plasmid in α-syn PFF-treated condition. Projections verify the proximity of the internalized α-syn fibrils; GFP (green), α-syn (red), DAPI (blue). Scale bar 30 μm. D Four images per cell culture were analyzed (n = 3). Quantifications of α-syn PFFs fluorescent-positive puncta were performed using ImageJ (ComDet plug-in). E Western blot analysis of primary striatal astrocyte lysates transfected with scramble and AnxA2 siRNA under basal and PFF-treated conditions. Anti-Lrrk2, anti-α-syn, and anti-AnxA2 antibodies have been employed. F, G, H Quantification of band intensity was performed using ImageJ and normalized by β-actin (n = 3). Statistical analysis in D was made by Kruskal-Wallis test followed by Dunn’s multiple comparisons test. Statistical analysis in F, G, and H was performed with an unpaired t test or one-way ANOVA followed by Tukey’s multiple comparisons test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001
Fig. 6 Investigation of AnxA2 function in astrocyte-mediated α-syn phagocytic clearance. A Unlabeled α-syn PFFs have been applied to primary astrocytes for 24 h. Projections verify the proximity of the internalized α-syn fibrils and endogenous AnxA2. Cell cytoskeleton (F-actin, cyano), α-syn (red), AnxA2 (green), DAPI (blue). Scale bar 20 μm. B Schematic outline of the experimental setup. Cells were transfected using 3xFlag-GFP-encoding plasmid together with scramble or AnxA2 siRNA. 48 h post transfection cells were treated with 0.5 μM sonicated α-syn PFFs for 24 h and imaged using confocal microscopy. C Representative images of primary striatal astrocytes transfected with scramble or Anxa2 siRNA together with a GFP-encoding plasmid in α-syn PFF-treated condition. Projections verify the proximity of the internalized α-syn fibrils; GFP (green), α-syn (red), DAPI (blue). Scale bar 30 μm. D Four images per cell culture were analyzed (n = 3). Quantifications of α-syn PFFs fluorescent-positive puncta were performed using ImageJ (ComDet plug-in). E Western blot analysis of primary striatal astrocyte lysates transfected with scramble and AnxA2 siRNA under basal and PFF-treated conditions. Anti-Lrrk2, anti-α-syn, and anti-AnxA2 antibodies have been employed. F, G, H Quantification of band intensity was performed using ImageJ and normalized by β-actin (n = 3). Statistical analysis in D was made by Kruskal-Wallis test followed by Dunn’s multiple comparisons test. Statistical analysis in F, G, and H was performed with an unpaired t test or one-way ANOVA followed by Tukey’s multiple comparisons test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001CC BY · PubMed Central
Fig. 7 Analysis of AnxA2 function in G2019S primary striatal astrocytes at endogenous level. A Western blot analysis of primary striatal astrocyte lysates under PFF-treated and basal conditions using anti-Lrrk2, anti-α-syn, and anti-AnxA2 antibodies. B, C, D Quantification of band intensity was performed using ImageJ and normalized by β-actin (n = 4). E Representative images of Lrrk2+/+ and Lrrk2GS/GS astrocytes treated or not with α-syn PFFs and stained with anti-AnxA2 (green), anti-α-syn (red), F-actin (cyano), and cell nuclei with DAPI (blue). MLi-2-treated Lrrk2GS/GS astrocytes are shown in the bottom panels. Scale bar 20 μm. Insets show a close-up of α-syn inclusions and re-localized AnxA2. F, G Eight images per cell culture were analyzed (n = 3). Quantifications of Anxa2 puncta and AnxA2-α-syn PFFs proximity were performed using ImageJ. Statistical analysis in F and G was made by Kruskal-Wallis test followed by Dunn’s multiple comparisons test. Statistical analysis in B, C, and D was performed with unpaired t test or one-way ANOVA followed by Tukey’s multiple comparisons test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001
Fig. 7 Analysis of AnxA2 function in G2019S primary striatal astrocytes at endogenous level. A Western blot analysis of primary striatal astrocyte lysates under PFF-treated and basal conditions using anti-Lrrk2, anti-α-syn, and anti-AnxA2 antibodies. B, C, D Quantification of band intensity was performed using ImageJ and normalized by β-actin (n = 4). E Representative images of Lrrk2+/+ and Lrrk2GS/GS astrocytes treated or not with α-syn PFFs and stained with anti-AnxA2 (green), anti-α-syn (red), F-actin (cyano), and cell nuclei with DAPI (blue). MLi-2-treated Lrrk2GS/GS astrocytes are shown in the bottom panels. Scale bar 20 μm. Insets show a close-up of α-syn inclusions and re-localized AnxA2. F, G Eight images per cell culture were analyzed (n = 3). Quantifications of Anxa2 puncta and AnxA2-α-syn PFFs proximity were performed using ImageJ. Statistical analysis in F and G was made by Kruskal-Wallis test followed by Dunn’s multiple comparisons test. Statistical analysis in B, C, and D was performed with unpaired t test or one-way ANOVA followed by Tukey’s multiple comparisons test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001CC BY · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Research works printed but not linked: 71

  1. Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M (1997) Alpha-synuclein in Lewy bodies. Nature 388:839–840 · DOI · Paper page
  2. Wakabayashi K, Hayashi S, Yoshimoto M, Kudo H, Takahashi H (2000) NACP/alpha-synuclein-positive filamentous inclusions in astrocytes and oligodendrocytes of Parkinson’s disease brains. Acta Neuropathol 99:14–20 · DOI
  3. Terada S, Ishizu H, Yokota O, Tsuchiya K, Nakashima H, Ishihara T, Fujita D, Ueda K et al (2003) Glial involvement in diffuse Lewy body disease. Acta Neuropathol 105:163–169 · DOI
  4. Croisier E, Graeber MB (2006) Glial degeneration and reactive gliosis in alpha-synucleinopathies: the emerging concept of primary gliodegeneration. Acta Neuropathol 112:517–530 · DOI
  5. Rostami J, Fotaki G, Sirois J, Mzezewa R, Bergstrom J, Essand M, Healy L, Erlandsson A (2020) Astrocytes have the capacity to act as antigen-presenting cells in the Parkinson’s disease brain. J Neuroinflammation 17:119 · DOI · Paper page
  6. Biskup S, Moore DJ, Rea A, Lorenz-Deperieux B, Coombes CE, Dawson VL, Dawson TM, West AB (2007) Dynamic and redundant regulation of LRRK2 and LRRK1 expression. BMC Neurosci 8:102 · DOI
  7. Booth HDE, Hirst WD, Wade-Martins R (2017) The role of astrocyte dysfunction in Parkinson’s disease pathogenesis. Trends Neurosci 40:358–370 · DOI · Paper page
  8. Tolosa E, Vila M, Klein C, Rascol O (2020) LRRK2 in Parkinson disease: challenges of clinical trials. Nat Rev Neurol 16:97–107 · DOI
  9. Alessi DR, Sammler E (2018) LRRK2 kinase in Parkinson’s disease. Science 360:36–37 · DOI · Paper page
  10. Martin I, Dawson VL, Dawson TM (2011) Recent advances in the genetics of Parkinson’s disease. Annu Rev Genomics Hum Genet 12:301–325 · DOI
  11. Cogo S, Manzoni C, Lewis PA, Greggio E (2020) Leucine-rich repeat kinase 2 and lysosomal dyshomeostasis in Parkinson disease. J Neurochem 152:273–283 · DOI
  12. Cookson MR (2017) Mechanisms of Mutant LRRK2 Neurodegeneration. Adv Neurobiol 14:227–239 · DOI
  13. Chen J, Chen Y, Pu J (2018) Leucine-rich repeat kinase 2 in Parkinson’s disease: updated from pathogenesis to potential therapeutic target. Eur Neurol 79:256–265 · DOI
  14. Schapansky J, Khasnavis S, DeAndrade MP, Nardozzi JD, Falkson SR, Boyd JD, Sanderson JB, Bartels T et al (2018) Familial knockin mutation of LRRK2 causes lysosomal dysfunction and accumulation of endogenous insoluble alpha-synuclein in neurons. Neurobiol Dis 111:26–35 · DOI
  15. Tong Y, Giaime E, Yamaguchi H, Ichimura T, Liu Y, Si H, Cai H, Bonventre JV et al (2012) Loss of leucine-rich repeat kinase 2 causes age-dependent bi-phasic alterations of the autophagy pathway. Mol Neurodegener 7:2 · DOI
  16. Bieri G, Brahic M, Bousset L, Couthouis J, Kramer NJ, Ma R, Nakayama L, Monbureau M et al (2019) LRRK2 modifies alpha-syn pathology and spread in mouse models and human neurons. Acta Neuropathol 137:961–980 · DOI
  17. Sanyal A, DeAndrade MP, Novis HS, Lin S, Chang J, Lengacher N, Tomlinson JJ, Tansey MG et al (2020) Lysosome and inflammatory defects in GBA1-mutant astrocytes are normalized by LRRK2 inhibition. Mov Disord 35:760–773 · DOI
  18. Hartlova A, Herbst S, Peltier J, Rodgers A, Bilkei-Gorzo O, Fearns A, Dill BD, Lee H et al (2018) LRRK2 is a negative regulator of Mycobacterium tuberculosis phagosome maturation in macrophages. EMBO J 37:e98694 · DOI
  19. Lee H, Flynn R, Sharma I, Haberman E, Carling PJ, Nicholls FJ, Stegmann M, Vowles J et al (2020) LRRK2 is recruited to phagosomes and co-recruits RAB8 and RAB10 in human pluripotent stem cell-derived macrophages. Stem Cell Reports 14:940–955 · DOI
  20. Loov C, Hillered L, Ebendal T, Erlandsson A (2012) Engulfing astrocytes protect neurons from contact-induced apoptosis following injury. PLoS One 7:e33090 · DOI
  21. Loov C, Mitchell CH, Simonsson M, Erlandsson A (2015) Slow degradation in phagocytic astrocytes can be enhanced by lysosomal acidification. Glia 63:1997–2009 · DOI
  22. Loria F, Vargas JY, Bousset L, Syan S, Salles A, Melki R, Zurzolo C (2017) Alpha-synuclein transfer between neurons and astrocytes indicates that astrocytes play a role in degradation rather than in spreading. Acta Neuropathol 134:789–808 · DOI
  23. Ferrazza R, Cogo S, Melrose H, Bubacco L, Greggio E, Guella G, Civiero L, Plotegher N (2016) LRRK2 deficiency impacts ceramide metabolism in brain. Biochem Biophys Res Commun 478:1141–1146 · DOI
  24. Civiero L, Cirnaru MD, Beilina A, Rodella U, Russo I, Belluzzi E, Lobbestael E, Reyniers L et al (2015) Leucine-rich repeat kinase 2 interacts with p21-activated kinase 6 to control neurite complexity in mammalian brain. J Neurochem 135:1242–1256 · DOI
  25. Longo F, Mercatelli D, Novello S, Arcuri L, Brugnoli A, Vincenzi F, Russo I, Berti G et al (2017) Age-dependent dopamine transporter dysfunction and Serine129 phospho-alpha-synuclein overload in G2019S LRRK2 mice. Acta Neuropathol Commun 5:22 · DOI
  26. Davis AA, Temple S (1994) A self-renewing multipotential stem cell in embryonic rat cerebral cortex. Nature 372:263–266 · DOI
  27. Johe KK, Hazel TG, Muller T, Dugich-Djordjevic MM, McKay RD (1996) Single factors direct the differentiation of stem cells from the fetal and adult central nervous system. Genes Dev 10:3129–3140 · DOI
  28. Ravin R, Hoeppner DJ, Munno DM, Carmel L, Sullivan J, Levitt DL, Miller JL, Athaide C et al (2008) Potency and fate specification in CNS stem cell populations in vitro. Cell Stem Cell 3:670–680 · DOI
  29. Sollvander S, Nikitidou E, Gallasch L, Zysk M, Soderberg L, Sehlin D, Lannfelt L, Erlandsson A (2018) The Abeta protofibril selective antibody mAb158 prevents accumulation of Abeta in astrocytes and rescues neurons from Abeta-induced cell death. J Neuroinflammation 15:98 · DOI
  30. Civiero L, Vancraenenbroeck R, Belluzzi E, Beilina A, Lobbestael E, Reyniers L, Gao F, Micetic I et al (2012) Biochemical characterization of highly purified leucine-rich repeat kinases 1 and 2 demonstrates formation of homodimers. PLoS One 7:e43472 · DOI
  31. Pivato M, De Franceschi G, Tosatto L, Frare E, Kumar D, Aioanei D, Brucale M, Tessari I et al (2012) Covalent alpha-synuclein dimers: chemico-physical and aggregation properties. PLoS One 7:e50027 · DOI
  32. Kim Y, Ho SO, Gassman NR, Korlann Y, Landorf EV, Collart FR, Weiss S (2008) Efficient site-specific labeling of proteins via cysteines. Bioconjug Chem 19:786–791 · DOI
  33. Hurbain I, Romao M, Bergam P, Heiligenstein X, Raposo G (2017) Analyzing lysosome-related organelles by electron microscopy. Methods Mol Biol 1594:43–71 · DOI
  34. Carraro M, Checchetto V, Sartori G, Kucharczyk R, di Rago JP, Minervini G, Franchin C, Arrigoni G et al (2018) High-conductance channel formation in yeast mitochondria is mediated by F-ATP synthase e and g subunits. Cell Physiol Biochem 50:1840–1855 · DOI
  35. Gustafsson G, Lindstrom V, Rostami J, Nordstrom E, Lannfelt L, Bergstrom J, Ingelsson M, Erlandsson A (2017) Alpha-synuclein oligomer-selective antibodies reduce intracellular accumulation and mitochondrial impairment in alpha-synuclein exposed astrocytes. J Neuroinflammation 14:241 · DOI
  36. Westerlund M, Belin AC, Anvret A, Bickford P, Olson L, Galter D (2008) Developmental regulation of leucine-rich repeat kinase 1 and 2 expression in the brain and other rodent and human organs: implications for Parkinson’s disease. Neuroscience 152:429–436 · DOI
  37. Giesert F, Hofmann A, Burger A, Zerle J, Kloos K, Hafen U, Ernst L, Zhang J et al (2013) Expression analysis of Lrrk1, Lrrk2 and Lrrk2 splice variants in mice. PLoS One 8:e63778 · DOI
  38. Hawrysh PJ, Buck LT (2019) Mitochondrial matrix pH acidifies during anoxia and is maintained by the F1Fo-ATPase in anoxia-tolerant painted turtle cortical neurons. FEBS Open Bio 9:571–581 · DOI
  39. Lerch S, Ritz S, Bley K, Messerschmidt C, Weiss CK, Musyanovych A, Landfester K, Mailander V (2015) Nanoprobing the acidification process during intracellular uptake and trafficking. Nanomedicine 11:1585–1596 · DOI
  40. Repetto G, del Peso A, Zurita JL (2008) Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nat Protoc 3:1125–1131 · DOI
  41. Pinsino A, Russo R, Bonaventura R, Brunelli A, Marcomini A, Matranga V (2015) Titanium dioxide nanoparticles stimulate sea urchin immune cell phagocytic activity involving TLR/p38 MAPK-mediated signalling pathway. Sci Rep 5:14492 · DOI
  42. Shen K, Sidik H, Talbot WS (2016) The rag-ragulator complex regulates lysosome function and phagocytic flux in microglia. Cell Rep 14:547–559 · DOI
  43. Scharf B, Clement CC, Wu XX, Morozova K, Zanolini D, Follenzi A, Larocca JN, Levon K et al (2012) Annexin A2 binds to endosomes following organelle destabilization by particulate wear debris. Nat Commun 3:755 · DOI
  44. Li R, Tan S, Yu M, Jundt MC, Zhang S, Wu M (2015) Annexin A2 regulates autophagy in pseudomonas aeruginosa infection through the Akt1-mTOR-ULK1/2 signaling pathway. J Immunol 195:3901–3911 · DOI
  45. Moreau K, Ghislat G, Hochfeld W, Renna M, Zavodszky E, Runwal G, Puri C, Lee S et al (2015) Transcriptional regulation of annexin A2 promotes starvation-induced autophagy. Nat Commun 6:8045 · DOI
  46. Morozova K, Sridhar S, Zolla V, Clement CC, Scharf B, Verzani Z, Diaz A, Larocca JN et al (2015) Annexin A2 promotes phagophore assembly by enhancing Atg16L(+) vesicle biogenesis and homotypic fusion. Nat Commun 6:5856 · DOI
  47. Bustos V, Pulina MV, Bispo A, Lam A, Flajolet M, Gorelick FS, Greengard P (2017) Phosphorylated presenilin 1 decreases beta-amyloid by facilitating autophagosome-lysosome fusion. Proc Natl Acad Sci U S A 114:7148–7153 · DOI
  48. de Graauw M, Cao L, Winkel L, van Miltenburg MH, le Devedec SE, Klop M, Yan K, Pont C et al (2014) Annexin A2 depletion delays EGFR endocytic trafficking via cofilin activation and enhances EGFR signaling and metastasis formation. Oncogene 33:2610–2619 · DOI
  49. Stukes S, Coelho C, Rivera J, Jedlicka AE, Hajjar KA, Casadevall A (2016) The membrane phospholipid binding protein annexin A2 promotes phagocytosis and nonlytic exocytosis of cryptococcus neoformans and impacts survival in fungal infection. J Immunol 197:1252–1261 · DOI
  50. Zhang Y, Sloan SA, Clarke LE, Caneda C, Plaza CA, Blumenthal PD, Vogel H, Steinberg GK et al (2016) Purification and characterization of progenitor and mature human astrocytes reveals transcriptional and functional differences with mouse. Neuron 89:37–53 · DOI · Paper page
  51. Fell MJ, Mirescu C, Basu K, Cheewatrakoolpong B, DeMong DE, Ellis JM, Hyde LA, Lin Y et al (2015) MLi-2, a potent, selective, and centrally active compound for exploring the therapeutic potential and safety of LRRK2 kinase inhibition. J Pharmacol Exp Ther 355:397–409 · DOI
  52. Recasens A, Dehay B (2014) Alpha-synuclein spreading in Parkinson’s disease. Front Neuroanat 8:159 · DOI
  53. Luk KC, Kehm VM, Zhang B, O’Brien P, Trojanowski JQ, Lee VMY (2012) Intracerebral inoculation of pathological α-synuclein initiates a rapidly progressive neurodegenerative α-synucleinopathy in mice. J Exp Med 209:975–986 · DOI
  54. Desplats P, Lee HJ, Bae EJ, Patrick C, Rockenstein E, Crews L, Spencer B, Masliah E et al (2009) Inclusion formation and neuronal cell death through neuron-to-neuron transmission of -synuclein. Proc Natl Acad Sci 106:13010–13015 · DOI · Paper page
  55. Lee HJ, Patel S, Lee SJ (2005) Intravesicular localization and exocytosis of alpha-synuclein and its aggregates. J Neurosci 25:6016–6024 · DOI
  56. Reyes JF, Olsson TT, Lamberts JT, Devine MJ, Kunath T, Brundin P (2015) A cell culture model for monitoring α-synuclein cell-to-cell transfer. Neurobiol Dis 77:266–275 · DOI
  57. Danzer KM, Ruf WP, Putcha P, Joyner D, Hashimoto T, Glabe C, Hyman BT, McLean PJ (2011) Heat-shock protein 70 modulates toxic extracellular alpha-synuclein oligomers and rescues trans-synaptic toxicity. FASEB J 25:326–336 · DOI
  58. Zhao Y, Keshiya S, Atashrazm F, Gao J, Ittner LM, Alessi DR, Halliday GM, Fu Y et al (2018) Nigrostriatal pathology with reduced astrocytes in LRRK2 S910/S935 phosphorylation deficient knockin mice. Neurobiol Dis 120:76–87 · DOI
  59. Lee JH, Han JH, Kim H, Park SM, Joe EH, Jou I (2019) Parkinson’s disease-associated LRRK2-G2019S mutant acts through regulation of SERCA activity to control ER stress in astrocytes. Acta Neuropathol Commun 7:68 · DOI
  60. Lara Ordonez AJ, Fernandez B, Fdez E, Romo-Lozano M, Madero-Perez J, Lobbestael E, Baekelandt V, Aiastui A et al (2019) RAB8, RAB10 and RILPL1 contribute to both LRRK2 kinase-mediated centrosomal cohesion and ciliogenesis deficits. Hum Mol Genet 28:3552–3568 · DOI · Paper page
  61. Booth HDE, Wessely F, Connor-Robson N, Rinaldi F, Vowles J, Browne C, Evetts SG, Hu MT et al (2019) RNA sequencing reveals MMP2 and TGFB1 downregulation in LRRK2 G2019S Parkinson’s iPSC-derived astrocytes. Neurobiol Dis 129:56–66 · DOI
  62. Hyttinen JMT, Amadio M, Viiri J, Pascale A, Salminen A, Kaarniranta K (2014) Clearance of misfolded and aggregated proteins by aggrephagy and implications for aggregation diseases. Ageing Res Rev 18:16–28 · DOI
  63. Baptista MA, Dave KD, Frasier MA, Sherer TB, Greeley M, Beck MJ, Varsho JS, Parker GA et al (2013) Loss of leucine-rich repeat kinase 2 (LRRK2) in rats leads to progressive abnormal phenotypes in peripheral organs. PLoS One 8:e80705 · DOI
  64. Walter J, Bolognin S, Antony PMA, Nickels SL, Poovathingal SK, Salamanca L, Magni S, Perfeito R et al (2019) Neural stem cells of Parkinson’s disease patients exhibit aberrant mitochondrial morphology and functionality. Stem Cell Reports 12:878–889 · DOI
  65. Hipolito VEB, Diaz JA, Tandoc KV, Oertlin C, Ristau J, Chauhan N, Saric A, McLaughlan S et al (2019) Enhanced translation expands the endo-lysosome size and promotes antigen presentation during phagocyte activation. PLoS Biol 17:e3000535 · DOI
  66. Mayran N, Parton RG, Gruenberg J (2003) Annexin II regulates multivesicular endosome biogenesis in the degradation pathway of animal cells. EMBO J 22:3242–3253 · DOI
  67. Grieve AG, Moss SE, Hayes MJ (2012) Annexin A2 at the interface of actin and membrane dynamics: a focus on its roles in endocytosis and cell polarization. Int J Cell Biol 2012:852430 · DOI
  68. Eberhard DA, Brown MD, VandenBerg SR (1994) Alterations of annexin expression in pathological neuronal and glial reactions. Immunohistochemical localization of annexins I, II (p36 and p11 subunits), IV, and VI in the human hippocampus. Am J Pathol 145:640–649
  69. Juul-Madsen K, Qvist P, Bendtsen KL, Langkilde AE, Vestergaard B, Howard KA, Dehesa-Etxebeste M, Paludan SR et al (2020) Size-selective phagocytic clearance of fibrillar alpha-synuclein through conformational activation of complement Receptor 4. J Immunol 204:1345–1361 · DOI
  70. Zhang Q, Xu Y, Lee J, Jarnik M, Wu X, Bonifacino JS, Shen J, Ye Y (2020) A myosin-7B-dependent endocytosis pathway mediates cellular entry of alpha-synuclein fibrils and polycation-bearing cargos. Proc Natl Acad Sci U S A 117:10865–10875 · DOI
  71. Rodriguez L, Marano MM, Tandon A (2018) Import and export of misfolded alpha-synuclein. Front Neurosci 12:344 · DOI · Paper page

Cited by

Sorted by citations

See all 100

Metadata

Authors
Streubel-Gallasch, L., Giusti, V., Sandre, M., Tessari, I., Plotegher, N., Giusto, E., Masato, A., Iovino, L., Battisti, I., Arrigoni, G., Shimshek, D., Greggio, E., Tremblay, M.-E., Bubacco, L., Erlandsson, A., & Civiero, L.
Venue
Molecular Neurobiology
Publisher
Springer Science and Business Media LLC
Published
2021-02-24
Volume
58
Issue
7
Pages
3119-3140
ISSN
  • 0893-7648
  • 1559-1182
DOI
10.1007/s12035-021-02327-8
PMC
PMC8257537
Cited by
100
References
22
Language
en
Licence
Funders
  • Università degli Studi di Padova
  • Italian Ministry of Health
  • ParkinsonFörbundet
  • Dementia Association Foundation
  • Swedish Brain Foundation
  • Swedish Research Council
  • Ahlen Foundation
  • Stiftelsen Olle Engkvist Byggmästare
  • Torsten Söderbergs Stiftelse

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review