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Cited by 549Neuroinflammation and Neurodegeneration MechanismsParkinson's Disease Mechanisms and TreatmentsAutophagy in Disease and Therapy

Microglia clear neuron-released α-synuclein via selective autophagy and prevent neurodegeneration

Open access
Choi, Insup, Zhang, Yuanxi, Seegobin, Steven P., Pruvost, Mathilde, Wang, Qian, Purtell, Kerry, Zhang, Bin, Yue, Zhenyu·2020·Nature Communications
DOI 10.1038/s41467-020-15119-wPMC PMC7069981

Published version of a preprint

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Abstract

Microglia maintain brain homeostasis by removing neuron-derived components such as myelin and cell debris. The evidence linking microglia to neurodegenerative diseases is growing; however, the precise mechanisms remain poorly understood. Herein, we report a neuroprotective role for microglia in the clearance of neuron-released α-synuclein. Neuronal α-synuclein activates microglia, which in turn engulf α-synuclein into autophagosomes for degradation via selective autophagy (termed synucleinphagy). Synucleinphagy requires the presence of microglial Toll-like receptor 4 (TLR4), which induces transcriptional upregulation of p62/SQSTM1 through the NF-κB signaling pathway. Induction of p62, an autophagy receptor, is necessary for the formation of α-synuclein/ubiquitin-positive puncta that are degraded by autophagy. Finally, disruption of microglial autophagy in mice expressing human α-synuclein promotes the accumulation of misfolded α-synuclein and causes midbrain dopaminergic neuron degeneration. Our study thus identifies a neuroprotective function of microglia in the clearance of α-synuclein via TLR4-NF-κB-p62 mediated synucleinphagy.

AI snapshot

Written from the paper’s full text.

TL;DR

Microglia engulf neuron-released α-synuclein and degrade it through selective autophagy, termed synucleinphagy, via TLR4–NF-κB–p62/SQSTM1 signaling. Disrupting microglial autophagy in mice expressing human α-synuclein increases misfolded α-synuclein and causes midbrain dopaminergic neuron degeneration.

Key findings

  • Neuron-released human α-synuclein (hα-Syn) is engulfed by microglia in vivo in AAV9-hα-Syn-injected mice and hα-Syn-Tg mice, accompanied by microglial activation and detection of hα-Syn in isolated microglia.
  • Ingested hα-Syn forms ubiquitin-, p62-, and GFP–LC3-positive puncta that peak at 6 h and decline by 24 h; clearance is blocked by SAR405, Bafilomycin A1, or Chloroquine, enhanced by Torin1, and abolished in Atg7- or Atg14-deficient microglia and p62-KO microglia.
  • hα-Syn-induced p62 upregulation requires TLR4 and NF-κB: Tlr4-KO, TAK-242, or ML-120B suppresses p62 induction and hα-Syn/ubiquitin puncta; hα-Syn prefers TLR4 over TLR2 for NF-κB activation in HEK293T reporter assays and does not induce TLR4 endocytosis or IRF3 signaling like LPS.
  • Microglia-specific Atg7 deficiency in hα-Syn mice increases detergent-insoluble hα-Syn, p-S129 α-synuclein-positive structures, and decreases TH-positive and Nissl-positive cells in the SNpc at 7 months after tamoxifen; hα-Syn-Tg mice alone showed no dopaminergic degeneration up to 12 months.

Methods

  • Mouse models included AAV9-GFP or AAV9-hα-Syn injection into SNpc, Thy-1 hα-Syn transgenic mice, Cx3cr1CreER-IRES-Eyfp, Atg7flox/flox, Atg14flox/flox, p62-KO, Tlr4-KO, and GFP–LC3 mice; primary microglia, adult microglia isolation, and HEK293T NF-κB-luciferase TLR assays were used.
  • Main analyses included RT-qPCR, western blot, immunostaining with confocal 3D reconstruction, electron microscopy, co-immunoprecipitation, brain fractionation into detergent-soluble/insoluble fractions, flow cytometry, and stereological counting of TH/Nissl-positive SNpc neurons.
  • Statistics used unpaired two-tailed Student’s t test or Mann–Whitney U test, one-way ANOVA with Newman–Keuls post hoc, or two-way ANOVA with Bonferroni post hoc; values are mean ± SEM.

Limitations

  • The paper states that the exact mechanism by which α-synuclein enters microglia remains unresolved, the specific E3 ligase for α-synuclein in microglia is unidentified, and the significance of other TLRs requires future study.
  • Evident from the text: mechanistic dissection relies heavily on cultured primary microglia and recombinant hα-Syn, while in vivo evidence uses overexpression models (AAV or Thy-1 transgenic hα-Syn) rather than endogenous α-synuclein; no human tissue or clinical data are presented.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1038/s41467-020-15119-w

Cites

74 of 74 references verified0 citation errors · 0 not checkable

Statistics

51 tests recomputed · 0 with the reported and recomputed p on opposite sides of .05

Claims the paper makes

52 of 73 cited sources checked · 24 supported · 4 flagged · accuracy not yet measured

  • Premisecitation

    TLR4 is required for increased inflammation in response to various forms of α-synuclein.

    “TLR4 was required for NF-κB activation, and increased inflammation and phagocytic activity in response to various forms of α-synuclein40,41,56.”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites [40] Fellner, L. et al. Toll-like receptor 4 is required for alpha-synuclein dependent activation of microglia and astroglia. Glia 61, 349–360 (2013). source
    • cites [41] Rannikko, E. H., Weber, S. S. & Kahle, P. J. Exogenous alpha-synuclein induces toll-like receptor 4 dependent inflammatory responses in astrocytes. BMC Neurosci. 16, 57 (2015). source

      The Results section reports that α-synuclein induced inflammatory-gene expression in control astrocytes but not significantly in TLR4-knockout astrocytes. However, the Discussion says the study did not determine which α-synuclein species were involved, so it does not establish the claim for various forms. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [56] Stefanova, N. et al. Toll-like receptor 4 promotes alpha-synuclein clearance and survival of nigral dopaminergic neurons. Am. J. Pathol. 179, 954–963 (2011). source

      The Results section reports the opposite for an inflammatory measure: α-synuclein-overexpressing mice lacking TLR4 had increased midbrain TNF-α and a proinflammatory astroglial response. The Discussion supports a role for TLR4 in α-synuclein phagocytosis, but not the attributed requirement for increased inflammation. (LLM, conf=0.86)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
  • Premisecitation

    TLR4 is required for increased phagocytic activity in response to various forms of α-synuclein.

    “TLR4 was required for NF-κB activation, and increased inflammation and phagocytic activity in response to various forms of α-synuclein40,41,56.”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites [40] Fellner, L. et al. Toll-like receptor 4 is required for alpha-synuclein dependent activation of microglia and astroglia. Glia 61, 349–360 (2013). source
    • cites [41] Rannikko, E. H., Weber, S. S. & Kahle, P. J. Exogenous alpha-synuclein induces toll-like receptor 4 dependent inflammatory responses in astrocytes. BMC Neurosci. 16, 57 (2015). source

      The Results section, “Primary astrocytes internalize αSYN from the medium in a TLR4-independent manner” (page unavailable), reports no considerable difference in α-synuclein internalization between TLR4-positive and TLR4-knockout astrocytes, including across the tested variants. The paper supports TLR4-dependent inflammatory signaling, but says α-synuclein uptake is TLR4-independent rather than requiring TLR4. (LLM, conf=0.86)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [56] Stefanova, N. et al. Toll-like receptor 4 promotes alpha-synuclein clearance and survival of nigral dopaminergic neurons. Am. J. Pathol. 179, 954–963 (2011). source
  • Findingown data

    Treatment with Torin1, an mTOR inhibitor that stimulates autophagy, facilitates clearance of hα-Syn/ubiquitin-positive puncta.

    “By contrast, treatment with Torin1, an inhibitor of mTOR that stimulates autophagy37, facilitated the clearance of puncta (Fig. 2g).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [37] Liu, Q. S. et al. Discovery of 1-(4-(4-Propionylpiperazin-1-yl)-3-(trifluoromethyl)phenyl)-9-(quinolin-3-yl)benzo[h][1,6]naphthyridin-2(1H)-one as a highly potent, selective mammalian target of rapamycin (mTOR) inhibitor for the treatment of cancer. J. Med. Chem. 53, 7146–7155 (2010). source

      The Results and discussion identify Torin1 as an inhibitor of cellular mTOR activity. The Introduction says mTOR regulates autophagy, but our copy does not establish that Torin1 stimulates autophagy. The citing sentence attributes puncta clearance to its own Fig. 2g, not to this source. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
  • Premisecitation

    Evidence for macroautophagy degradation of α-synuclein is extremely limited.

    “However, the evidence for macroautophagy (hereafter referred to as autophagy) degradation of α-synuclein is extremely limited24,25.”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [24] Lin, Y. C., Kuo, H. C., Wang, J. S. & Lin, W. W. Regulation of inflammatory response by 3-methyladenine involves the coordinative actions on akt and glycogen synthase kinase 3 beta rather than autophagy. J. Immunol. 189, 4154–4164 (2012). source
    • cites [25] Wu, Y. T. et al. Dual role of 3-methyladenine in modulation of autophagy via different temporal patterns of inhibition on class I and III phosphoinositide 3-kinase. J. Biol. Chem. 285, 10850–10861 (2010). source

      The ‘Dual Role of 3-Methyladenine in Modulation of Autophagy’ section reports autophagic flux and degradation of long-lived proteins, not α-synuclein. Searches for α-synuclein and synuclein found no passage assessing whether evidence for its macroautophagic degradation is limited. Page numbers were not available in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
  • Premisecitation

    α-synuclein can be secreted by neurons as a result of cellular stress or injury, or as a response to stimulation.

    “α-synuclein can be secreted by neurons as a result of cellular stress or injury, or as a response to stimulation11–13.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [11] Emmanouilidou, E. et al. Assessment of alpha-synuclein secretion in mouse and human brain parenchyma. PLoS One 6, e22225 (2011). source

      The Introduction and Results and Discussion report physiological release of α-synuclein into brain interstitial fluid, supporting neuronal secretion. The study detected it in patients with severe brain injury, but does not establish injury or cellular stress as the cause of secretion; it also does not show secretion in response to stimulation. Its discussion of probe insertion argues against neuronal membrane damage explaining the measured release. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [12] Jang, A. et al. Non-classical exocytosis of alpha-synuclein is sensitive to folding states and promoted under stress conditions. J. Neurochem. 113, 1263–1274 (2010). source
    • cites [13] Yamada, K. & Iwatsubo, T. Extracellular alpha-synuclein levels are regulated by neuronal activity. Mol. Neurodegener. 13, 9 (2018). source

      The Discussion reports that stressed cells release more α-synuclein, and the Results show that increased neuronal activity increases its release. The paper does not establish injury as a cause of α-synuclein secretion; the Background instead notes that release can occur independently of cell death. Page numbers are unavailable in our copy. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
  • Classified as probably not a substantial claim. 27
  • May combine two claims. 19

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    TLR4-NF-κB signaling induces selective autophagy after pathogen invasion.

    “TLR4-NF-κB signaling was previously shown to induce selective autophagy after pathogen invasion52–54.”
    Quote in the paperNot a substantial claim3 of 3 sources checked
    • cites [52] Shi, C. S. & Kehrl, J. H. MyD88 and Trif target Beclin 1 to trigger autophagy in macrophages. J. Biol. Chem. 283, 33175–33182 (2008). source

      The ‘TLRs Induce Autophagy in Murine’ section reports that TLR4 ligands induced autophagosome formation in macrophages. The Discussion identifies MyD88, Trif and Beclin 1 in the proposed mechanism; although it mentions TLR-mediated NF-κB activation, it does not show that NF-κB induces the autophagy. The experiments also do not establish selective autophagy after pathogen invasion. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [53] Watson, R. O., Manzanillo, P. S. & Cox, J. S. Extracellular M. tuberculosis DNA targets bacteria for autophagy by activating the host DNA-sensing pathway. Cell 150, 803–815 (2012). source

      In Results, under “Cytosolic DNA Sensing Mediates Targeting of M. tuberculosis to Autophagosomes” (page unavailable), the paper reports that autophagy targeting remained normal in macrophages deficient in TLR signaling. It instead identifies cytosolic DNA sensing through STING as necessary for selective targeting of M. tuberculosis; it does not show induction by TLR4–NF-κB signaling. (LLM, conf=0.86)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [54] Xu, Y. et al. Toll-like receptor 4 is a sensor for autophagy associated with innate immunity. Immunity 27, 135–144 (2007). source

      The Results show that LPS induces autophagy through TLR4, and the Discussion identifies a TRIF-dependent pathway involving RIP1 and p38 MAPK. The paper also reports colocalization of mycobacterial phagosomes with autophagosomes, but it does not establish NF-κB as the mediator or characterize the response as selective autophagy after pathogen invasion. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    α-synuclein can penetrate cytoplasmic membrane through endocytosis-independent process.

    “A number of studies demonstrated that α-synuclein can enter the cells or penetrate cytoplasmic membrane through endocytosis-independent process61–63.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [61] Ahn, K. J., Paik, S. R., Chung, K. C. & Kim, J. Amino acid sequence motifs and mechanistic features of the membrane translocation of alpha-synuclein. J. Neurochem. 97, 265–279 (2006). source
    • cites [62] Lee, H. J. et al. Assembly-dependent endocytosis and clearance of extracellular alpha-synuclein. Int. J. Biochem. Cell Biol. 40, 1835–1849 (2008). source
    • cites [63] Park, J. Y. et al. On the mechanism of internalization of alpha-synuclein into microglia: roles of ganglioside GM1 and lipid raft. J. Neurochem. 110, 400–411 (2009). source

      The Discussion describes extracellular monomeric α-synuclein entering microglia via GM1-mediated endocytosis. It says this uptake is independent of clathrin, caveolae and dynamin, but lipid raft-dependent; independence from those pathways is not independence from endocytosis. (LLM, conf=0.88)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    Atg7 encodes an E1-like enzyme essential in the ubiquitin-like conjugation systems required for autophagy, and Atg14 encodes a positive regulator of VPS34.

    “Atg7 encodes an E1-like enzyme essential in the ubiquitin-like conjugation systems required for autophagy, and Atg14 encodes a positive regulator of VPS34 (refs. 38,39).”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [38] Komatsu, M. et al. Impairment of starvation-induced and constitutive autophagy in Atg7-deficient mice. J. Cell Biol. 169, 425–434 (2005). source
    • cites [39] Matsunaga, K. et al. Autophagy requires endoplasmic reticulum targeting of the PI3-kinase complex via Atg14L. J. Cell Biol. 190, 511–521 (2010). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Aggregated α-synuclein is a major component in intraneuronal inclusions known as Lewy bodies associated with Parkinson's disease and dementia with Lewy body.

    “Aggregated α-synuclein is a major component in intraneuronal inclusions known as Lewy bodies (LB) associated with neurodegenerative diseases, such as Parkinson’s disease (PD) and dementia with Lewy body (DLB)5,6.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [5] Baba, M. et al. Aggregation of alpha-synuclein in Lewy bodies of sporadic Parkinson’s disease and dementia with Lewy bodies. Am. J. Pathol. 152, 879–884 (1998).
    • cites [6] Toledo, J. B. et al. Pathological alpha-synuclein distribution in subjects with coincident Alzheimer’s and Lewy body pathology. Acta Neuropathol. 131, 393–409 (2016). source

      The Introduction describes α-synuclein in the form of Lewy body–related pathology in Parkinson’s disease and discusses that pathology in dementia with Lewy bodies. It does not specify that Lewy bodies are intraneuronal inclusions or that aggregated α-synuclein is a major component of them. (LLM, conf=0.82)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    α-synuclein can trigger TLR4 or TLR2 activation in cultured glia, leading to an increase in inflammatory gene transcription.

    “Previous reports showed α-synuclein can trigger TLR4 or TLR2 activation in cultured glia, which leads to an increase in inflammatory gene transcription19,40,41.”
    Quote in the paperNot a substantial claim3 of 3 sources checked
    • cites [19] Kim, C. et al. Neuron-released oligomeric alpha-synuclein is an endogenous agonist of TLR2 for paracrine activation of microglia. Nat. Commun. 4, 1562 (2013). source
    • cites [40] Fellner, L. et al. Toll-like receptor 4 is required for alpha-synuclein dependent activation of microglia and astroglia. Glia 61, 349–360 (2013). source

      The Results (page unavailable) show that α-synuclein treatment caused TLR4-dependent NF-κB nuclear translocation and increased inflammatory cytokine release in cultured murine glia. The paper measured cytokine release, not an increase in inflammatory gene transcription; its experiments address TLR4 rather than TLR2. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [41] Rannikko, E. H., Weber, S. S. & Kahle, P. J. Exogenous alpha-synuclein induces toll-like receptor 4 dependent inflammatory responses in astrocytes. BMC Neurosci. 16, 57 (2015). source

  • Classified as probably not a substantial claim.

    Premisecitation

    p62 is an autophagy receptor that recognizes and interacts with ubiquitinated cargo to initiate selective autophagy.

    “p62/SQSTM1, an autophagy receptor34”
    • Also stated in: § p62 mediates sequestration and degradation of α-synuclein · boosted
      “p62 is known to recognize and interact with ubiquitinated cargo to initiate selective autophagy34.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [34] Lamark, T., Kirkin, V., Dikic, I. & Johansen, T. NBR1 and p62 as cargo receptors for selective autophagy of ubiquitinated targets. Cell Cycle 8, 1986–1990 (2009). source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    TLR4-LPS binding induces endocytosis of the TLR4-LPS complex and subsequent expression of Ifnb1 through activation of IRF3.

    “TLR4-LPS binding induces endocytosis of the TLR4-LPS complex and subsequent expression of interferon-beta 1 (Ifnb1) through activation of interferon regulatory factor 3 (IRF3)43,44.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [43] Sakaguchi, S. et al. Essential role of IRF-3 in lipopolysaccharide-induced interferon-beta gene expression and endotoxin shock. Biochem. Bioph. Res. Commun. 306, 860–866 (2003). source
    • cites [44] Zanoni, I. et al. CD14 controls the LPS-induced endocytosis of Toll-like receptor 4. Cell 147, 868–880 (2011). source

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    Torin1 is an inhibitor of mTOR that stimulates autophagy.

    “Torin1, an inhibitor of mTOR that stimulates autophagy37”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [37] Liu, Q. S. et al. Discovery of 1-(4-(4-Propionylpiperazin-1-yl)-3-(trifluoromethyl)phenyl)-9-(quinolin-3-yl)benzo[h][1,6]naphthyridin-2(1H)-one as a highly potent, selective mammalian target of rapamycin (mTOR) inhibitor for the treatment of cancer. J. Med. Chem. 53, 7146–7155 (2010). source

      The Abstract identifies Torin1 as an mTOR inhibitor, and the Introduction says mTOR regulates autophagy. Our copy does not state that Torin1 stimulates autophagy; the autophagy claim is therefore unsupported by this source. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Previous studies suggest α-synuclein is degraded by macroautophagy, chaperone-mediated autophagy, and the proteasome.

    “Previous studies suggest that α-synuclein is degraded by macroautophagy, chaperone-mediated autophagy, and the proteasome20–23.”
    Quote in the paperNot a substantial claim3 of 4 sources checked
    • cites [20] Cuervo, A. M., Stefanis, L., Fredenburg, R., Lansbury, P. T. & Sulzer, D. Impaired degradation of mutant alpha-synuclein by chaperone-mediated autophagy. Science 305, 1292–1295 (2004). source
    • cites [21] Tofaris, G. K. et al. Ubiquitin ligase Nedd4 promotes alpha-synuclein degradation by the endosomal-lysosomal pathway. Proc. Natl Acad. Sci. USA 108, 17004–17009 (2011). source
    • cites [22] Vogiatzi, T., Xilouri, M., Vekrellis, K. & Stefanis, L. Wild type alpha-synuclein is degraded by chaperone-mediated autophagy and macroautophagy in neuronal cells. J. Biol. Chem. 283, 23542–23556 (2008). source
    • cites [23] Webb, J. L., Ravikumar, B., Atkins, J., Skepper, J. N. & Rubinsztein, D. C. alpha-synuclein is degraded by both autophagy and the proteasome. J. Biol. Chem. 278, 25009–25013 (2003). source

      The Discussion reports that α-synuclein is degraded by the proteasome and autophagy in inducible PC12 cells. The paper describes autophagy through the formation of autophagosomes, consistent with macroautophagy, but our copy does not address chaperone-mediated autophagy; a search for that term found no supporting passage. (LLM, conf=0.87)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    p62 has a role in the clearance of intra-cytosolic Listeria.

    “p62 has a role in the clearance of intra-cytosolic Shigella and Listeria55.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [55] Mostowy, S. et al. p62 and NDP52 proteins target intracytosolic Shigella and Listeria to different autophagy pathways. J. Biol. Chem. 286, 26987–26995 (2011). source

      The Discussion says p62 targets the Listeria ActA mutant to an autophagy pathway, supporting a role in its clearance. The section on p62 and NDP52 recruitment specifies that wild-type Listeria avoids p62 recognition, so the claim is too broad without the ActA-mutant qualification. Page numbers are not available in our copy. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    α-synuclein can enter cells through endocytosis-independent process.

    “A number of studies demonstrated that α-synuclein can enter the cells or penetrate cytoplasmic membrane through endocytosis-independent process61–63.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [61] Ahn, K. J., Paik, S. R., Chung, K. C. & Kim, J. Amino acid sequence motifs and mechanistic features of the membrane translocation of alpha-synuclein. J. Neurochem. 97, 265–279 (2006). source
    • cites [62] Lee, H. J. et al. Assembly-dependent endocytosis and clearance of extracellular alpha-synuclein. Int. J. Biochem. Cell Biol. 40, 1835–1849 (2008). source
    • cites [63] Park, J. Y. et al. On the mechanism of internalization of alpha-synuclein into microglia: roles of ganglioside GM1 and lipid raft. J. Neurochem. 110, 400–411 (2009). source

      The Discussion reports that high-dose α-synuclein uptake can be temperature-independent and that monomeric uptake into microglia is independent of clathrin, caveolae and dynamin. But it describes the studied route as lipid-raft-dependent endocytosis, not as independent of endocytosis altogether. No page number is available in our copy. (LLM, conf=0.85)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    TLR2 plays a role in microglia in response to α-synuclein.

    “TLR2 also plays a role in neurons in response to α-synuclein in addition to microglia19,57,58.”
    Quote in the paperNot a substantial claim3 of 3 sources checked
    • cites [19] Kim, C. et al. Neuron-released oligomeric alpha-synuclein is an endogenous agonist of TLR2 for paracrine activation of microglia. Nat. Commun. 4, 1562 (2013). source
    • cites [57] Dzamko, N. et al. Toll-like receptor 2 is increased in neurons in Parkinson’s disease brain and may contribute to alpha-synuclein pathology. Acta Neuropathol. 133, 303–319 (2017). source
    • cites [58] Kim, C. et al. Antagonizing neuronal Toll-like receptor 2 prevents synucleinopathy by activating autophagy. Cell Rep. 13, 771–782 (2015). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    p62 has a role in the clearance of intra-cytosolic Shigella.

    “p62 has a role in the clearance of intra-cytosolic Shigella and Listeria55.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [55] Mostowy, S. et al. p62 and NDP52 proteins target intracytosolic Shigella and Listeria to different autophagy pathways. J. Biol. Chem. 286, 26987–26995 (2011). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    TLR2 plays a role in neurons in response to α-synuclein.

    “TLR2 also plays a role in neurons in response to α-synuclein in addition to microglia19,57,58.”
    Quote in the paperNot a substantial claim3 of 3 sources checked
    • cites [19] Kim, C. et al. Neuron-released oligomeric alpha-synuclein is an endogenous agonist of TLR2 for paracrine activation of microglia. Nat. Commun. 4, 1562 (2013). source

      The Results section on TLR2 upregulation reports TLR2 labeling in pyramidal neurons, but the α-synuclein-related increase it describes is in microglia. The in vivo Results show that α-synuclein expressed by neurons activates microglia through TLR2; they do not establish that TLR2 mediates a neuronal response to α-synuclein. Page numbers are unavailable in our copy. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [57] Dzamko, N. et al. Toll-like receptor 2 is increased in neurons in Parkinson’s disease brain and may contribute to alpha-synuclein pathology. Acta Neuropathol. 133, 303–319 (2017). source

      The Introduction and Discussion report neuronal TLR2 expression and show that activating neuronal TLR2 increases α-synuclein protein. They do not show that neurons activate TLR2 in response to α-synuclein; the experiments activated TLR2 with an agonist, so the direction implied by “in response to” is not established. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [58] Kim, C. et al. Antagonizing neuronal Toll-like receptor 2 prevents synucleinopathy by activating autophagy. Cell Rep. 13, 771–782 (2015). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Autophagy-lysosome system is a converging pathogenic pathway of PD.

    “emerging human genomic studies suggest the autophagy–lysosome system as a converging pathogenic pathway of PD64”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [64] Chang, D. et al. A meta-analysis of genome-wide association studies identifies 17 new Parkinson’s disease risk loci. Nat. Genet. 49, 1511–1516 (2017). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Many PD-related genes such as PINK1, PARKIN, GBA, and LRRK2 play a role in autophagy-related pathways in glial cells.

    “many PD-related genes such as PINK1, PARKIN, GBA, and LRRK2, were shown to play a role in autophagy-related pathways in glial cells65.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [65] Gan-Or, Z., Dion, P. A. & Rouleau, G. A. Genetic perspective on the role of the autophagy-lysosome pathway in Parkinson disease. Autophagy 11, 1443–1457 (2015). source

      The Introduction and the sections on PARK2/PINK1, GBA, and LRRK2 link these PD-related genes to autophagy or lysosomal pathways. The passages do not establish that these roles were shown specifically in glial cells, so the cell-type qualifier is unsupported. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Findingown data

    Microglia-engulfed α-synuclein is sequestered by autophagosomes and degraded by the autophagy-lysosome pathway.

    “Thus, we concluded that microglia engulf extracellular α-syn protein, which is sequestered by autophagosomes and degraded by the autophagy–lysosome pathway.”
    • Also stated in: § Abstract · plain
      “which in turn engulf α-synuclein into autophagosomes for degradation via selective autophagy (termed synucleinphagy).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The two mouse models used (AAV9-hα-Syn and hα-Syn-Tg) faithfully model neuron-released α-synuclein relevant to human synucleinopathies.

  • May combine two claims.

    Findingown data

    hα-Syn treatment does not significantly alter overall autophagy, as total LC3-II levels change little, and does not alter other autophagy receptors Ndp52, Optineurin, and Nbr1.

    “total LC3-II levels changed little after hα-Syn treatment (Supplementary Fig. 4i), suggesting no significant alteration of overall autophagy.”
    • Also stated in: § p62 mediates sequestration and degradation of α-synuclein · hedged
      “other autophagy receptors such as Ndp52, Optineurin, and Nbr1 were not altered (Supplementary Fig. 4i), suggesting that hα-Syn selectively induces p62 expression without affecting other autophagy receptors.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    hα-Syn treatment induces p62 protein and mRNA upregulation in cultured microglia.

    “We observed that hα-Syn treatment caused an increase in p62 protein up to 9 h (Fig. 3a).”
    • Also stated in: § p62 mediates sequestration and degradation of α-synuclein · plain
      “p62 mRNA levels peaked at 3 h, declined at 6 h, and returned to baseline at 9 h post treatment (Fig. 3b)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Actinomycin D inhibits p62 induction and suppresses formation of hα-Syn/ubiquitin-positive puncta in microglia.

    “Actinomycin D, a blocker of mRNA translation, inhibited p62 induction when added before hα-Syn treatment (Supplementary Fig. 4g) and suppressed the formation of hα-Syn/ubiquitin-positive puncta (Supplementary fig. 4h).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Uptake of hα-Syn by microglia is not inhibited by cytochalasin D or Dynasore; rather, these treatments increase uptake.

    “Neither cytochalasin D, a blocker of actin polymerization necessary for phagocytosis, nor Dynasore, a GTPase inhibitor that suppresses both clathrin-dependent and independent endocytosis, inhibited the uptake of hα-Syn protein but rather it increased the uptake (Supplementary Figs. 4b, 5a, 5b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    p38 inhibitors SB202190 and SB203580 do not significantly suppress p62 induction or reduce hα-Syn/ubiquitin-positive puncta after hα-Syn treatment.

    “only ML-120B significantly suppressed p62 induction and reduced the number of hα-Syn/ubiquitin-positive puncta after hα-Syn treatment (Fig. 5b, c)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Disruption of microglial autophagy in mice expressing human α-synuclein promotes accumulation of misfolded α-synuclein and causes midbrain dopaminergic neuron degeneration.

    “Finally, disruption of microglial autophagy in mice expressing human α-synuclein promotes the accumulation of misfolded α-synuclein and causes midbrain dopaminergic neuron degeneration.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The two mouse models used (AAV9-hα-Syn and hα-Syn-Tg) faithfully model neuron-released α-synuclein relevant to human synucleinopathies.

  • May combine two claims.

    Findingown data

    hα-Syn treatment does not induce Ifnb1 expression or IRF3 activation in microglia, unlike LPS.

    “LPS, but not hα-Syn, induced Ifnb1 expression (Fig. 4h) and increased levels of p-IFR3 (S396), a marker of IRF3 activation (Fig. 4i), indicating the absence of IRF3 signaling in hα-Syn-treated microglia.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    TAK-242 treatment impairs p62 induction and reduces hα-Syn/ubiquitin-positive puncta following hα-Syn treatment.

    “Similar to Tlr4-KO microglia, TAK-242-treated cells showed impaired p62 induction (Fig. 4d) as well as a reduction in the number of hα-Syn/ubiquitin-positive puncta following hα-Syn treatment (Fig. 4e).”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    TAK-242 is an inhibitor of TLR4 signaling by selectively binding to Cys747 of TLR4 cytoplasmic tail that disrupts its interaction with adaptor molecules TIRAP and TRAM.

    “TAK-242, an inhibitor of TLR4 signaling by selectively binding to Cys747 of TLR4 cytoplasmic tail that disrupts its interaction with adaptor molecules TIRAP and TRAM42”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [42] Matsunaga, N., Tsuchimori, N., Matsumoto, T. & Ii, M. TAK-242 (resatorvid), a small-molecule inhibitor of Toll-like receptor (TLR) 4 signaling, binds selectively to TLR4 and interferes with interactions between TLR4 and its adaptor molecules. Mol. Pharm. 79, 34–41 (2011). source

  • May combine two claims.

    Interpretationown data

    The hα-Syn-TLR4 interaction stimulates p62 expression without causing TLR4 receptor endocytosis, distinctively from LPS-TLR4 signaling.

    “The above observations suggest that the hα-Syn-TLR4 interaction stimulates p62 expression without causing TLR4 receptor endocytosis distinctively from LPS-TLR4 signaling.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Recent evidence demonstrates a strong selectivity for autophagy.

    “Recent evidence demonstrates a strong selectivity for autophagy28.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [28] Stolz, A., Ernst, A. & Dikic, I. Cargo recognition and trafficking in selective autophagy. Nat. Cell Biol. 16, 495–501 (2014). source

  • May combine two claims.

    Interpretationreasoning

    The results uncover a microglia-specific α-synuclein degradation pathway termed synucleinphagy that regulates α-synuclein homeostasis in the CNS.

    “Thus, our results uncover a microglia-specific α-synuclein degradation pathway termed synucleinphagy that regulates α-synuclein homeostasis in the CNS.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Direct evidence for autophagy in selective degradation of α-synuclein is entirely lacking.
    • Rests on an assumption: Whether microglia take part in the degradation of α-synuclein remains unclear.
    • Rests on an assumption: The exact mechanism for the clearance of internalized α-synuclein remains unclear.

  • Classified as probably not a substantial claim.

    Premisecitation

    Previous evidence suggested prion-like cell-to-cell transmission of α-synuclein.

    “Previous evidence suggested “prion-like” cell-to-cell transmission of α-synuclein9,10.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [9] Kordower, J. H., Chu, Y., Hauser, R. A., Freeman, T. B. & Olanow, C. W. Lewy body-like pathology in long-term embryonic nigral transplants in Parkinson’s disease. Nat. Med. 14, 504–506 (2008). source
    • cites [10] Li, J. Y. et al. Lewy bodies in grafted neurons in subjects with Parkinson’s disease suggest host-to-graft disease propagation. Nat. Med. 14, 501–503 (2008). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Autophagy is a bulk degradation pathway responsible for clearance of protein aggregates and damaged cellular organelles.

    “Autophagy is a bulk degradation pathway responsible for the clearance of protein aggregates and damaged cellular organelles26,27.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [26] Mizushima, N., Yoshimori, T. & Ohsumi, Y. The role of Atg proteins in autophagosome formation. Annu. Rev. Cell Dev. Biol. 27, 107–132 (2011). source
    • cites [27] Yamamoto, A. & Yue, Z. Autophagy and its normal and pathogenic states in the brain. Annu. Rev. Neurosci. 37, 55–78 (2014). source

  • Classified as probably not a substantial claim.

    Premisecitation

    SAR405 is an inhibitor of VPS34, a class III phosphatidylinositol 3-kinase required for autophagosome membrane synthesis.

    “SAR405, an inhibitor of VPS34, which is a class III phosphatidylinositol 3-kinase required for autophagosome membrane synthesis36”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [36] Ronan, B. et al. A highly potent and selective Vps34 inhibitor alters vesicle trafficking and autophagy. Nat. Chem. Biol. 10, 1013 (2014). source

  • May combine two claims.

    Interpretationown data

    Synucleinphagy requires TLR4, which induces transcriptional upregulation of p62 through the NF-κB signaling pathway.

    “Synucleinphagy requires the presence of microglial Toll-like receptor 4 (TLR4), which induces transcriptional upregulation of p62/SQSTM1 through the NF-κB signaling pathway.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Increased α-synuclein level due to multiplication of SNCA alleles is causal to Parkinson's disease.

    “Increased α-synuclein level due to the multiplication of SNCA alleles is causal to PD7,8.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [7] Chartier-Harlin, M. C. et al. alpha-synuclein locus duplication as a cause of familial Parkinson’s disease. Lancet 364, 1167–1169 (2004). source
    • cites [8] Singleton, A. B. et al. alpha-synuclein locus triplication causes Parkinson’s disease. Science 302, 841–841 (2003). source

  • May combine two claims.

    Interpretationown data

    Ingested α-synuclein in microglia is sequestered by autophagosomes for degradation, mediated by TLR4-NF-κB signaling through upregulation of p62/SQSTM1.

    “We document that ingested α-synuclein in microglia is sequestered by autophagosomes for degradation, which is mediated by TLR4-NF-κB signaling through upregulation of the autophagy receptor, p62/SQSTM1.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Direct evidence for autophagy in selective degradation of α-synuclein is entirely lacking.
    • Rests on an assumption: Whether microglia take part in the degradation of α-synuclein remains unclear.
    • Rests on an assumption: The exact mechanism for the clearance of internalized α-synuclein remains unclear.

  • May combine two claims.

    Interpretationown data

    Oligomeric p62 directly binds and recruits hα-Syn into autophagosomes.

    “supporting that oligomeric p62 directly binds and recruits hα-Syn into autophagosomes.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The high molecular weight p62 species observed by co-immunoprecipitation is oligomeric p62.

  • May combine two claims.

    Interpretationreasoning

    Neurons lack the TLR4-NF-κB pathway, likely explaining why they show no change in autophagosomes or p62 after α-synuclein treatment.

    “likely due to lack of the TLR4-NF-κB pathway.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    In the NF-κB-luciferase HEK293T system, Pam3CSK4 selectively increases NF-κB activity in cells expressing TLR2 and TLR2 combinations.

    “Application of LPS, Pam3CSK4, and recombinant Flagellin peptides selectively increased NF-κB activity in TLR4 complex, TLR2 and TLR2 combinations, and TLR5, respectively (Supplementary Fig. 6).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    In the NF-κB-luciferase HEK293T system, Flagellin selectively increases NF-κB activity in cells expressing TLR5.

    “Application of LPS, Pam3CSK4, and recombinant Flagellin peptides selectively increased NF-κB activity in TLR4 complex, TLR2 and TLR2 combinations, and TLR5, respectively (Supplementary Fig. 6).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    Data are representative of at least three independent experiments unless indicated.

    “Data are representative of at least three independent experiments unless indicated.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductown data

    Stereological counting of TH-positive or Nissl-stained cell bodies in the SNpc using the stated parameters yielded a coefficient of error <10% throughout the analysis.

    “These parameters yielded a coefficient of error <10% throughout the analysis.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    A portion of extracellular α-synuclein interacts with TLR4 (or TLR2) triggering NF-κB signaling.

    “our study supports a model that a portion of extracellular α-synuclein interacts with TLR4 (or TLR2), which then triggers NF-κB signaling, while a majority of α-synuclein penetrates cytoplasmic membrane or enters microglia through TLR4 endocytosis-independent manner.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Data are representative of at least three independent experiments.

    “Data are representative of at least three independent experiments.”
    Quote in the paperSubstantial claim
43 of 43

  • The paper relies on this without support.

    The high molecular weight p62 species observed by co-immunoprecipitation is oligomeric p62.

    “supporting that oligomeric p62 directly binds and recruits hα-Syn into autophagosomes.”

  • The paper relies on this without support.

    Direct evidence for autophagy in selective degradation of α-synuclein is entirely lacking.

    “Direct evidence for autophagy in selective degradation of α-synuclein is entirely lacking.”

  • The paper relies on this without support.

    Whether microglia take part in the degradation of α-synuclein remains unclear.

    “whether or not microglia, the prototypical scavenger cell in the brain, take part in the degradation of α-synuclein remains unclear.”

  • The paper relies on this without support.

    The exact mechanism for the clearance of internalized α-synuclein remains unclear.

    “However, the exact mechanism for the clearance of internalized α-synuclein remains unclear.”

  • The paper relies on this without support.

    The two mouse models used (AAV9-hα-Syn and hα-Syn-Tg) faithfully model neuron-released α-synuclein relevant to human synucleinopathies.

    “we employed two mouse models, both of which express wild-type (WT) human α-synuclein (hα-Syn) in a neuron-specific manner.”
5 of 5

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?

  • In animal models of PD and in PD patients, misfolded α-synuclein is released from injured neurons into extracellular fluid.

    “In animal models of PD and in PD patients, misfolded α-synuclein was found to be released from injured neurons into the extracellular fluid.51,61,62”

    From Role of neuroinflammation in neurodegeneration development (2023) · cites this paper as [62]

    The Introduction (page not provided) says neurons can secrete α-synuclein following stress or injury, and the Results (page not provided) report extracellular neuron-released α-synuclein in mouse models. The paper does not establish that the released protein was misfolded or report its release from injured neurons in PD patients. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

Statistics that don’t add up

Each reported p-value recomputed from its test statistic and degrees of freedom.

  • Inconsistent p-value: F(3, 6) = 11.46, p = .0003

    Minor

    The paper reports p = .0003 here; recomputing from the test statistic and its degrees of freedom gives p = .007. The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .0003 · recomputed p = .007

1 of 1

Statistics recomputed

Every reported test result, recomputed from its test statistic and degrees of freedom. A result shows as not adding up only once an independent check has confirmed it.

  • Doesn’t add upF(3, 6) = 11.46reported p = .0003 · recomputed p = .007

    “F(3, 6) = 11.46, p = .0003”

  • ConsistentF(5, 168) = 139.6reported p = 6.43e-58 · recomputed p < .001

    “F(5,168) = 139.6, p = 6.43e-58”

  • ConsistentF(5, 162) = 118.4reported p = 3.14e-52 · recomputed p < .001

    “F(5,162) = 118.4, p = 3.14e-52”

  • ConsistentF(3, 130) = 39.88reported p = 2.43e-18 · recomputed p < .001

    “F(3,130)=39.88, p = 2.43e-18”

  • ConsistentF(3, 8) = 13.44reported p = .0017 · recomputed p = .002

    “F(3,8) = 13.44, p = .0017”

  • ConsistentF(3, 20) = 35.43reported p = 3.39e-8 · recomputed p < .001

    “F(3,20) = 35.43, p = 3.39e-08”

  • ConsistentF(1, 139) = 272.5reported p = 1.44e-34 · recomputed p < .001

    “F(1,139)=272.5, p = 1.44e-34”

  • ConsistentF(1, 139) = 315.3reported p = 1.45e-37 · recomputed p < .001

    “F(1,139) = 315.3, p = 1.45e-37”

  • ConsistentF(1, 139) = 303.7reported p = 8.81e-37 · recomputed p < .001

    “F(1,139)=303.7, p = 8.81e-37”

  • ConsistentF(3, 16) = 3.381reported p = .0443 · recomputed p = .044

    “F(3,16) = 3.381, p = .0443”

  • ConsistentF(1, 16) = 43.09reported p = .00000651 · recomputed p < .001

    “F(1,16) = 43.09, p = 6.51e-06”

  • ConsistentF(3, 40) = 3.199reported p = .0334 · recomputed p = .033

    “F(3,40) = 3.199, p = .0334”

  • ConsistentF(3, 40) = 8.326reported p = .0002 · recomputed p < .001

    “F(3,40) = 8.326, p = .0002”

  • ConsistentF(1, 40) = 21.77reported p = .0000341 · recomputed p < .001

    “F(1,40) = 21.77, p = 3.41e-05”

  • ConsistentF(2, 174) = 12.56reported p = .00000803 · recomputed p < .001

    “F(2,174) = 12.56, p = 8.03e-06”

  • ConsistentF(2, 174) = 91.61reported p = 6.64e-28 · recomputed p < .001

    “F(2,174) = 91.61, p = 6.64e-28”

  • ConsistentF(1, 174) = 24.89reported p = .00000146 · recomputed p < .001

    “F(1,174) = 24.89, p = 1.46e-06”

  • ConsistentF(2, 92) = 156.7reported p = 2.35e-30 · recomputed p < .001

    “F(2,92) = 156.7, p = 2.35e-30”

  • ConsistentF(3, 20) = 35.43reported p = 3.39e-8 · recomputed p < .001

    “F(3,20)=35.43, p = 3.39e-08”

  • ConsistentF(3, 20) = 6.655reported p = .0027 · recomputed p = .003

    “F(3,20) = 6.655, p = .0027”

  • ConsistentF(3, 20) = 8.907reported p = .0006 · recomputed p < .001

    “F(3,20) = 8.907, p = .0006”

  • ConsistentF(3, 20) = 5.104reported p = .0087 · recomputed p = .009

    “F(3,20) = 5.104, p = .0087”

  • ConsistentF(3, 23) = 19.53reported p = .00000161 · recomputed p < .001

    “F(3,23) = 19.53, p = 1.61e-06”

  • ConsistentF(3, 23) = 4.643reported p = .0111 · recomputed p = .011

    “F(3,23) = 4.643, p = .0111”

  • ConsistentF(3, 23) = 12.09reported p = .0000592 · recomputed p < .001

    “F(3,23) = 12.09, p = 5.92e-05”

  • ConsistentF(3, 23) = 35.86reported p = 7.68e-9 · recomputed p < .001

    “F(3,23)=35.86, p = 7.68e-09”

  • ConsistentF(2, 18) = 15.1reported p = .0001 · recomputed p < .001

    “F(2,18) = 15.10, p = .0001”

  • ConsistentF(4, 24) = 9.196reported p = .0001 · recomputed p < .001

    “F(4,24) = 9.196, p = .0001”

  • ConsistentF(4, 24) = 4.53reported p = .0072 · recomputed p = .007

    “F(4,24) = 4.530, p = .0072”

  • ConsistentF(4, 24) = 0.8644reported p = .4994 · recomputed p = .499

    “F(4,24) = 0.8644, p = .4994”

  • ConsistentF(4, 24) = 1.369reported p = .2741 · recomputed p = .274

    “F(4,24) = 1.369, p = .2741”

  • ConsistentF(4, 24) = 5.372reported p = .0031 · recomputed p = .003

    “F(4,24) = 5.372, p = .0031”

  • ConsistentF(3, 12) = 3.75reported p = .0413 · recomputed p = .041

    “F(3,12) = 3.750, p = .0413”

  • ConsistentF(4, 160) = 56.21reported p = 1.53e-29 · recomputed p < .001

    “F(4,160) = 56.21, p = 1.53e-29”

  • ConsistentF(3, 40) = 20.7reported p = 2.95e-8 · recomputed p < .001

    “F(3,40) = 20.70, p = 2.95e-08”

  • ConsistentF(3, 40) = 69.43reported p = 6.55e-16 · recomputed p < .001

    “F(3,40) = 69.43, p = 6.55e-16”

  • ConsistentF(1, 40) = 69.57reported p = 2.74e-10 · recomputed p < .001

    “F(1,40) = 69.57, p = 2.74e-10”

  • ConsistentF(3, 40) = 10.12reported p = .0000432 · recomputed p < .001

    “F(3,40) = 10.12, p = 4.32e-05”

  • ConsistentF(3, 40) = 21.73reported p = 1.64e-8 · recomputed p < .001

    “F(3,40) = 21.73, p = 1.64e-08”

  • ConsistentF(1, 40) = 34.36reported p = 7.39e-7 · recomputed p < .001

    “F(1,40) = 34.36, p = 7.39e-07”

  • ConsistentF(3, 20) = 22.11reported p = .00000146 · recomputed p < .001

    “F(3,20) = 22.11, p = 1.46e-06”

  • ConsistentF(3, 20) = 11.88reported p = .0001 · recomputed p < .001

    “F(3,20) = 11.88, p = .0001”

  • ConsistentF(4, 80) = 19.63reported p = 2.74e-11 · recomputed p < .001

    “F(4,80) = 19.63, p = 2.74e-11”

  • ConsistentF(4, 80) = 27.31reported p = 2.66e-14 · recomputed p < .001

    “F(4,80) = 27.31, p = 2.66e-14”

  • ConsistentF(1, 80) = 47.95reported p = 9.9e-10 · recomputed p < .001

    “F(1,80) = 47.95, p = 9.90e-10”

  • ConsistentF(5, 126) = 6.07reported p = .0000454 · recomputed p < .001

    “F(5,126) = 6.070, p = 4.54e-05”

  • ConsistentF(5, 126) = 6.07reported p = .0000454 · recomputed p < .001

    “F(5,126) =6.070, p = 4.54e-05”

  • ConsistentF(1, 126) = 73.83reported p = 2.77e-14 · recomputed p < .001

    “F(1,126) = 73.83, p = 2.77e-14”

  • ConsistentF(3, 64) = 16.98reported p = 3.19e-8 · recomputed p < .001

    “F(3,64) = 16.98, p = 3.19e-08”

  • ConsistentF(3, 64) = 177.4reported p = 5.9e-31 · recomputed p < .001

    “F(3,64) = 177.4, p = 5.90e-31”

  • ConsistentF(1, 64) = 17.47reported p = .0000903 · recomputed p < .001

    “F(1,64) = 17.47, p = 9.03e-05”

51 of 51

Self-check

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

Each contradiction between two passages listed here was confirmed by an independent check.

  • Inconsistent residual degrees of freedom for the same two-way ANOVA

    Minor

    For Fig. 4a, the hα-Syn main effect is reported with 6 residual degrees of freedom, while the interaction and genotype effects from the same two-way ANOVA use 16. These effects should share the same residual degrees of freedom.

1 of 1

Cited works that were retracted

Works this paper cites that were later retracted, from the retraction notices publishers deposit. Major when the notice came out before this paper was published.

  • Cites a retracted work: 10.1038/ncb3192

    Info

    The cited work was retracted on 2024-03-04 (notice 10.1038/s41556-024-01383-1), not before this paper was published (2020-03-13).

    Cited work 10.1038/ncb3192 · retracted 2024-03-04 · notice 10.1038/s41556-024-01383-1 · this paper published 2020-03-13

1 of 1

Cites

1 retracted

Flagged first, then by citations

4 of the 78 references couldn’t be matched to a known paper.

Cited by

Sorted by citations

Related

Authors
Choi, I., Zhang, Y., Seegobin, S. P., Pruvost, M., Wang, Q., Purtell, K., Zhang, B., & Yue, Z.
Affiliations
Icahn School of Medicine at Mount Sinai (US)
Corresponding author
Yue, Zhenyu
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2020-03-13
Volume
11
Issue
1
Pages
1386-1386
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-020-15119-w
PMC
PMC7069981
OpenAlex
W3011306000
Semantic Scholar
0f4f3bc638feddb76ae7720b6dba5b2be88e755a
Cited by
549
References
78
Citations by year
2026: 71 · 2025: 112 · 2024: 97 · 2023: 75 · 2022: 100 · 2021: 69 · 2020: 25
FWCI
22.04
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion, cc-by)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
U.S. Department of Health & Human Services | NIH | National Institute of Neurological Disorders and Stroke (NS060123, P50NS094733)
Topics
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Health Sciences › Medicine › Neurology › Parkinson's Disease Mechanisms and Treatments
  • Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
Keywords
microglia, alpha-synuclein, selective autophagy, TLR4, NF-κB signaling, p62, neurodegeneration, dopaminergic neuron loss, neuroprotection, autophagy receptors, Parkinson's disease
MeSH terms
Animals, Autoantigens, Autophagy, Brain, Disease Models, Animal, Female, Humans, Mesencephalon, Mice, Inbred C57BL, Signal Transduction, NF-kappa B, Microglia, Mice, Knockout, Neurodegenerative Diseases, Toll-Like Receptor 4, Mice, alpha-Synuclein, HEK293 Cells, Dopaminergic Neurons

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.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-15.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-15.

Cited sources are checked by gpt-6-sol.

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.

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