CoChat
Sign Up
Cited by 147Parkinson's Disease Mechanisms and TreatmentsAutophagy in Disease and TherapyNuclear Receptors and Signaling

The DOI matches its Crossref record; 65 of the 65 references it cites were found in a registry or an index.

Age- and disease-dependent increase of the mitophagy marker phospho-ubiquitin in normal aging and Lewy body disease

Open access
Hou, Xu, Fiesel, Fabienne C., Truban, Dominika, Castanedes Casey, Monica, Lin, Wen-lang, Soto, Alexandra I., Tacik, Pawel, Rousseau, Linda G., Diehl, Nancy N., Heckman, Michael G., Lorenzo-Betancor, Oswaldo, Ferrer, Isidre, Arbelo, José M., Steele, John C., Farrer, Matthew J., Cornejo-Olivas, Mario, Torres, Luis, Mata, Ignacio F., Graff-Radford, Neill R., Wszolek, Zbigniew K., Ross, Owen A., Murray, Melissa E., Dickson, Dennis W., Springer, Wolfdieter·2018·Autophagy
DOI 10.1080/15548627.2018.1461294PMC PMC6372017
Open this paper in CoChat — free account

Abstract

Although exact causes of Parkinson disease (PD) remain enigmatic, mitochondrial dysfunction is increasingly appreciated as a key determinant of dopaminergic neuron susceptibility in both familial and sporadic PD. Two genes associated with recessive, early-onset PD encode the ubiquitin (Ub) kinase PINK1 and the E3 Ub ligase PRKN/PARK2/Parkin, which together orchestrate a protective mitochondrial quality control (mitoQC) pathway. Upon stress, both enzymes cooperatively identify and decorate damaged mitochondria with phosphorylated poly-Ub (p-S65-Ub) chains. This specific label is subsequently recognized by autophagy receptors that further facilitate mitochondrial degradation in lysosomes (mitophagy). Here, we analyzed human post-mortem brain specimens and identified distinct pools of p-S65-Ub-positive structures that partially colocalized with markers of mitochondria, autophagy, lysosomes and/or granulovacuolar degeneration bodies. We further quantified levels and distribution of the 'mitophagy tag' in 2 large cohorts of brain samples from normal aging and Lewy body disease (LBD) cases using unbiased digital pathology. Somatic p-S65-Ub structures independently increased with age and disease in distinct brain regions and enhanced levels in LBD brain were age- and Braak tangle stage-dependent. Additionally, we observed significant correlations of p-S65-Ub with LBs and neurofibrillary tangle levels in disease. The degree of co-existing p-S65-Ub signals and pathological PD hallmarks increased in the pre-mature stage, but decreased in the late stage of LB or tangle aggregation. Altogether, our study provides further evidence for a potential pathogenic overlap among different forms of PD and suggests that p-S65-Ub can serve as a biomarker for mitochondrial damage in aging and disease. ABBREVIATIONS: BLBD: brainstem predominant Lewy body disease; CCCP: carbonyl cyanide m-chlorophenyl hydrazone; DLB: dementia with Lewy bodies; DLBD: diffuse neocortical Lewy body disease; EOPD: early-onset Parkinson disease; GVB: granulovacuolar degeneration body; LB: Lewy body; LBD: Lewy body disease; mitoQC: mitochondrial quality control; nbM: nucleus basalis of Meynert; PD: Parkinson disease; PDD: Parkinson disease with dementia; p-S65-Ub: PINK1-phosphorylated serine 65 ubiquitin; SN: substantia nigra; TLBD: transitional Lewy body disease; Ub: ubiquitin.

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 · 10.1080/15548627.2018.1461294

Cites

65 of 65 references verified0 citation errors · 0 not checkable

Statistics

Figures

Figure 1. Morphology and distribution of p-S65-Ub-immunopositive structures. (A) Adjacent sections from LBD brains were stained with different p-S65-Ub antibodies at the indicated concentrations. Representative images of p-S65-Ub-immunopositive structures from various brain regions show the same 3 major patterns: granules (left panels), GVBs (middle panels) and beaded neurites (right panels). (B) Representative images of human brain sections from normal (left) and LBD cases (right) stained with p-S65-Ub antibody are shown. Chromogen used for the SN was HighDef Blue (blue) to distinguish the positive signal from dark brown neuromelanin in DA neurons. Conventional brown 3,3ʹ-diaminobenzidine was used as a chromogen for all other brain regions (hippocampus, amygdala, putamen, and nbM). A magnified image of the boxed area is shown to the right. Scale bars: 20 µm.
Figure 1. Morphology and distribution of p-S65-Ub-immunopositive structures. (A) Adjacent sections from LBD brains were stained with different p-S65-Ub antibodies at the indicated concentrations. Representative images of p-S65-Ub-immunopositive structures from various brain regions show the same 3 major patterns: granules (left panels), GVBs (middle panels) and beaded neurites (right panels). (B) Representative images of human brain sections from normal (left) and LBD cases (right) stained with p-S65-Ub antibody are shown. Chromogen used for the SN was HighDef Blue (blue) to distinguish the positive signal from dark brown neuromelanin in DA neurons. Conventional brown 3,3ʹ-diaminobenzidine was used as a chromogen for all other brain regions (hippocampus, amygdala, putamen, and nbM). A magnified image of the boxed area is shown to the right. Scale bars: 20 µm.CC BY-NC-ND · PubMed Central
Figure 2. Reduced or absent p-S65-Ub staining in brains from PRKN or PINK1 mutation carriers. (A) SN sections from controls, sporadic LBD and cases with PINK1 or PRKN mutations were stained with p-S65-Ub antibody. Representative images of stained sections show enhanced p-S65-Ub levels in sporadic PD, but reduced or absent signal in SN of PRKN or PINK1 mutant cases. A magnified image of the boxed area is shown to the right. (B) Quantification of p-S65-Ub signal in age-matched controls, sporadic LBD cases and mutation carriers (Kruskal-Wallis test, *p < 0.01). n = 17 for normal, n = 19 for sporadic PD, n = 7 for PD with PRKN mutation, and n = 2 for PD with PINK1 mutation.
Figure 2. Reduced or absent p-S65-Ub staining in brains from PRKN or PINK1 mutation carriers. (A) SN sections from controls, sporadic LBD and cases with PINK1 or PRKN mutations were stained with p-S65-Ub antibody. Representative images of stained sections show enhanced p-S65-Ub levels in sporadic PD, but reduced or absent signal in SN of PRKN or PINK1 mutant cases. A magnified image of the boxed area is shown to the right. (B) Quantification of p-S65-Ub signal in age-matched controls, sporadic LBD cases and mutation carriers (Kruskal-Wallis test, *p < 0.01). n = 17 for normal, n = 19 for sporadic PD, n = 7 for PD with PRKN mutation, and n = 2 for PD with PINK1 mutation.CC BY-NC-ND · PubMed Central
Figure 3. High-resolution imaging and ultrastructural examination of human brain tissue immunolabeled with p-S65-Ub. Human brain sections from LBD cases were multicolor labeled with antibodies against p-S65-Ub and (A) PPIF (mitochondrial marker), (B) CTSD (lysosomal marker) or (C) CSNK1D (GVB marker) and SSBP1 (mitochondrial marker). Representative Airyscan confocal images from SN and hippocampus are shown. p-S65-Ub-immunopositive granules partially colocalized with both mitochondrial and lysosomal markers; beaded neurites were co-labeled only with mitochondrial, but not lysosomal markers. GVBs were partially co-labeled with CSNK1D and mitochondrial markers. The colocalization of p-S65-Ub with the respective organelle markers is also demonstrated in the orthogonal (Ortho) view presented to the right. Scale bars: 5 µm. (D) Top: Mitochondria (black asterisks) are intensely labeled with p-S65-Ub on the outer membrane. Bottom: Some mitochondria (white asterisk) were found next to a neurofilament-like structure (solid arrow). (E, F) Top: At lower magnification, structures labeled (triangle) are consistent with GVBs seen at light microscopy. Bottom: At higher magnification, spherical double-membrane structures, possibly mitochondria, are found in the GVBs (white asterisk and circled by dashed line). Some labeled GVBs are adjacent to paired helical filaments (open arrow). Scale bars: 1 µm.
Figure 3. High-resolution imaging and ultrastructural examination of human brain tissue immunolabeled with p-S65-Ub. Human brain sections from LBD cases were multicolor labeled with antibodies against p-S65-Ub and (A) PPIF (mitochondrial marker), (B) CTSD (lysosomal marker) or (C) CSNK1D (GVB marker) and SSBP1 (mitochondrial marker). Representative Airyscan confocal images from SN and hippocampus are shown. p-S65-Ub-immunopositive granules partially colocalized with both mitochondrial and lysosomal markers; beaded neurites were co-labeled only with mitochondrial, but not lysosomal markers. GVBs were partially co-labeled with CSNK1D and mitochondrial markers. The colocalization of p-S65-Ub with the respective organelle markers is also demonstrated in the orthogonal (Ortho) view presented to the right. Scale bars: 5 µm. (D) Top: Mitochondria (black asterisks) are intensely labeled with p-S65-Ub on the outer membrane. Bottom: Some mitochondria (white asterisk) were found next to a neurofilament-like structure (solid arrow). (E, F) Top: At lower magnification, structures labeled (triangle) are consistent with GVBs seen at light microscopy. Bottom: At higher magnification, spherical double-membrane structures, possibly mitochondria, are found in the GVBs (white asterisk and circled by dashed line). Some labeled GVBs are adjacent to paired helical filaments (open arrow). Scale bars: 1 µm.CC BY-NC-ND · PubMed Central
Figure 4. p-S65-Ub cell density increases with advanced aging and Braak tangle stage in select brain regions in neurologically normal controls or in LBD. SN, hippocampus and amygdala from neurologically normal controls and LBD cases were stained with p-S65-Ub antibody. Correlations of p-S65-Ub cell density with age at death, Braak tangle stage and Thal amyloid phase was examined within either normal aging (A–C) or LBD cohort (D–F) (Spearman’s test of correlation, significance level: p < 0.01). In controls, n = 28 for SN, n = 28 for hippocampus, n = 22 for amygdala. Putamen and nbM are shown in Figure S4A-C. In LBD, n = 21 for SN, n = 28 for hippocampus, n = 24 for amygdala. Putamen and nbM are shown in Figure S4D-F.
Figure 4. p-S65-Ub cell density increases with advanced aging and Braak tangle stage in select brain regions in neurologically normal controls or in LBD. SN, hippocampus and amygdala from neurologically normal controls and LBD cases were stained with p-S65-Ub antibody. Correlations of p-S65-Ub cell density with age at death, Braak tangle stage and Thal amyloid phase was examined within either normal aging (A–C) or LBD cohort (D–F) (Spearman’s test of correlation, significance level: p < 0.01). In controls, n = 28 for SN, n = 28 for hippocampus, n = 22 for amygdala. Putamen and nbM are shown in Figure S4A-C. In LBD, n = 21 for SN, n = 28 for hippocampus, n = 24 for amygdala. Putamen and nbM are shown in Figure S4D-F.CC BY-NC-ND · PubMed Central
Figure 5. LBD brains have significantly higher p-S65-Ub levels compared to age-matched controls in SN and hippocampus. (A) p-S65-Ub-positive cell density from all 5 brain regions was compared between age-matched controls and LBD cases (Wilcoxon rank sum test, *p < 0.01). p-S65-Ub-positive cell density was also compared within (B) different age and (C) Braak tangle stage groups (Wilcoxon rank sum test followed by adjustment with Bonferroni correction, *p < 0.005). In controls, n = 20 for SN, n = 20 for hippocampus, n = 16 for amygdala, n = 16 for putamen, and n = 15 for nbM. In LBD, n = 21 for SN, n = 28 for hippocampus, n = 24 for amygdala, putamen, and nbM. All data are presented in scatter-plots showing the median and interquartile range.
Figure 5. LBD brains have significantly higher p-S65-Ub levels compared to age-matched controls in SN and hippocampus. (A) p-S65-Ub-positive cell density from all 5 brain regions was compared between age-matched controls and LBD cases (Wilcoxon rank sum test, *p < 0.01). p-S65-Ub-positive cell density was also compared within (B) different age and (C) Braak tangle stage groups (Wilcoxon rank sum test followed by adjustment with Bonferroni correction, *p < 0.005). In controls, n = 20 for SN, n = 20 for hippocampus, n = 16 for amygdala, n = 16 for putamen, and n = 15 for nbM. In LBD, n = 21 for SN, n = 28 for hippocampus, n = 24 for amygdala, putamen, and nbM. All data are presented in scatter-plots showing the median and interquartile range.CC BY-NC-ND · PubMed Central
Figure 6. Correlation and interaction of p-S65-Ub and SNCA pathology in LBD brains. (A) Lewy body density in SN, hippocampus and amygdala was compared between age-matched normal controls and LBD cases (Wilcoxon rank sum test, *p < 0.01). Data are presented in scatter-plots showing the median and interquartile range. (B) The correlation between p-S65-Ub and Lewy body density in LBD brains was examined, and significant correlations were observed in the amygdala (Spearman’s test of correlation, significance level: p < 0.01). In controls, n = 20 for SN, n = 20 for hippocampus, n = 16 for amygdala. In LBD, n = 21 for SN, n = 28 for hippocampus, n = 24 for amygdala. Putamen and nbM are shown in Figure S7A, B. (C) SN sections of LBD cases were double immunostained with p-S65-Ub and SNCA antibodies. Cells with different stages of SNCA-positive aggregations are shown. A magnified image of the boxed area is shown to the right. p-S65-Ub (green) colocalizes with SNCA (red) more in beaded neurites than in granules in soma. p-S65-Ub levels appear to decline with maturation of SNCA into LBs. For double labeling with p-S129-SNCA, see Figure S7C. (D) The spatial relationship of p-S65-Ub- and p-S129-SNCA-positive LB is shown in 3D surface rendering. Scale bars: 5 µm.
Figure 6. Correlation and interaction of p-S65-Ub and SNCA pathology in LBD brains. (A) Lewy body density in SN, hippocampus and amygdala was compared between age-matched normal controls and LBD cases (Wilcoxon rank sum test, *p < 0.01). Data are presented in scatter-plots showing the median and interquartile range. (B) The correlation between p-S65-Ub and Lewy body density in LBD brains was examined, and significant correlations were observed in the amygdala (Spearman’s test of correlation, significance level: p < 0.01). In controls, n = 20 for SN, n = 20 for hippocampus, n = 16 for amygdala. In LBD, n = 21 for SN, n = 28 for hippocampus, n = 24 for amygdala. Putamen and nbM are shown in Figure S7A, B. (C) SN sections of LBD cases were double immunostained with p-S65-Ub and SNCA antibodies. Cells with different stages of SNCA-positive aggregations are shown. A magnified image of the boxed area is shown to the right. p-S65-Ub (green) colocalizes with SNCA (red) more in beaded neurites than in granules in soma. p-S65-Ub levels appear to decline with maturation of SNCA into LBs. For double labeling with p-S129-SNCA, see Figure S7C. (D) The spatial relationship of p-S65-Ub- and p-S129-SNCA-positive LB is shown in 3D surface rendering. Scale bars: 5 µm.CC BY-NC-ND · PubMed Central
Figure 7. Correlation and interaction of p-S65-Ub and MAPT pathology in LBD brains. (A) tangle density in SN, hippocampus and amygdala is compared between age-matched normal controls and LBD cases (Wilcoxon rank sum test, *p < 0.01). Data are presented in scatter-plots with the median and interquartile range. (B) The correlation of p-S65-Ub with tangle density was examined in LBD, and significant correlations were observed in the hippocampus and amygdala (Spearman’s test of correlation, significance level: p < 0.01). In controls, n = 20 for SN, n = 20 for hippocampus, n = 16 for amygdala. In LBD, n = 21 for SN, n = 28 for hippocampus, n = 24 for amygdala. Putamen and nbM are shown in Figure S8A, B. (C) Hippocampal sections of LBD cases were double stained with p-S65-Ub and p-MAPT antibodies. A magnified image of the boxed area is shown to the right. Intracellular p-S65-Ub GVBs (green) are embedded in the p-MAPT network (red) labeled by late stage p-MAPT marker (PHF-1). For early stage p-MAPT marker CP13, see Figure S8C, D. p-S65-Ub levels appear to increase with maturation of tangles, but disappear in late-stage tangles. (D) The spatial relationship of p-S65-Ub and PHF-1 is shown in 3D surface (left) and maximum (right) rendering. Scale bars: 5 µm.
Figure 7. Correlation and interaction of p-S65-Ub and MAPT pathology in LBD brains. (A) tangle density in SN, hippocampus and amygdala is compared between age-matched normal controls and LBD cases (Wilcoxon rank sum test, *p < 0.01). Data are presented in scatter-plots with the median and interquartile range. (B) The correlation of p-S65-Ub with tangle density was examined in LBD, and significant correlations were observed in the hippocampus and amygdala (Spearman’s test of correlation, significance level: p < 0.01). In controls, n = 20 for SN, n = 20 for hippocampus, n = 16 for amygdala. In LBD, n = 21 for SN, n = 28 for hippocampus, n = 24 for amygdala. Putamen and nbM are shown in Figure S8A, B. (C) Hippocampal sections of LBD cases were double stained with p-S65-Ub and p-MAPT antibodies. A magnified image of the boxed area is shown to the right. Intracellular p-S65-Ub GVBs (green) are embedded in the p-MAPT network (red) labeled by late stage p-MAPT marker (PHF-1). For early stage p-MAPT marker CP13, see Figure S8C, D. p-S65-Ub levels appear to increase with maturation of tangles, but disappear in late-stage tangles. (D) The spatial relationship of p-S65-Ub and PHF-1 is shown in 3D surface (left) and maximum (right) rendering. Scale bars: 5 µm.CC BY-NC-ND · 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

Show all 65 references

Research works printed but not linked: 2

  1. Kosaka K, 1984, Clin Neuropathol, 3, 185
  2. van Elteren PH, 1960, Bull Inst Int Stat, 37, 351

Cited by

Sorted by citations

Items 101–147 of 147

Related

See all

Metadata

Authors
Hou, X., Fiesel, F. C., Truban, D., Castanedes Casey, M., Lin, W.-L., Soto, A. I., Tacik, P., Rousseau, L. G., Diehl, N. N., Heckman, M. G., Lorenzo-Betancor, O., Ferrer, I., Arbelo, J. M., Steele, J. C., Farrer, M. J., Cornejo-Olivas, M., Torres, L., Mata, I. F., Graff-Radford, N. R., Wszolek, Z. K., Ross, O. A., Murray, M. E., Dickson, D. W., & Springer, W.
Affiliations
  • Mayo Clinic in Florida (US)
  • Mayo Clinic (US)
  • Department of Neuroscience, Mayo Clinic, Jacksonville, FL, USA
  • Institut de Neuropatologia, Servei d’Anatomia Patològica, Hospital Universitari de Bellvitge, Hospitalet del Llobregat, Barcelona, Spain
  • Hospital Universitario Insular de Gran Canaria (ES)
  • Micronesian Health Study II, Tamuning, Guam
  • University of British Columbia (CA)
  • Northern Pacific Global Health Research Fellows Training Consortium, Bethesda, MD, USA
  • Instituto Nacional de Ciencias Neurológicas (PE)
  • University of Washington (US)
  • Department of Neurology, Mayo Clinic, Jacksonville, FL, USA
Corresponding author
Springer, Wolfdieter
ORCID
Venue
Autophagy
Publisher
Informa UK Limited
Published
2018-07-28
Volume
14
Issue
8
Pages
1404-1418
ISSN
  • 1554-8627 (ISSN-L)
  • 1554-8635
DOI
10.1080/15548627.2018.1461294
PMC
PMC6372017
OpenAlex
W2810603360
Cited by
147
References
65
Citations by year
2026: 18 · 2025: 28 · 2024: 16 · 2023: 19 · 2022: 17 · 2021: 13 · 2020: 25 · 2019: 10 · 2018: 1
FWCI
8.32
Type
journal article
Language
en
Open access
Licence
cc-by-nc-nd
Locations
  • Autophagy (publishedVersion, cc-by-nc-nd)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • Dipòsit Digital de la Universitat de Barcelona (Universitat de Barcelona) (acceptedVersion)
  • Acceda (Universidad de Las Palmas de Gran Canaria) (submittedVersion)
  • RECERCAT (Consorci de Serveis Universitaris de Catalunya) (submittedVersion, public-domain)
Funders
HHS | NIH | National Institute of Neurological Disorders and Stroke (R01 NS085070, U54 NS100693, R01 NS078086, P50 NS072187)
Topics
  • Health Sciences › Medicine › Clinical Neurology › Parkinson's Disease Mechanisms and Treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Autophagy in Disease and Therapy
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Receptors and Signaling
Keywords
mitophagy, Parkinson's disease, Lewy body disease, normal aging, mitochondrial quality control, PINK1, Parkin, ubiquitin phosphorylation, lysosomes, neurofibrillary tangles, biomarkers, dopaminergic neurons, mitochondrial dysfunction
MeSH terms
Aged, Aged, 80 and over, Aging, Brain, Female, HeLa Cells, Humans, Male, Middle Aged, Mutation, Phosphorylation, Phosphoserine, Protein Binding, Cohort Studies, Biomarkers, tau Proteins, Lewy Body Disease, Ubiquitin, alpha-Synuclein, Mitophagy

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