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Cited by 17Neuroinflammation and Neurodegeneration MechanismsPrion Diseases and Protein MisfoldingAlzheimer's disease research and treatments

Pharmacological modulation of TSPO in microglia/macrophages and neurons in a chronic neurodegenerative model of prion disease

Open access
Vicente-Rodríguez, Marta, Mancuso, Renzo, Peris-Yague, Alba, Simmons, Camilla, NIMA Consortium, Wlazly, Dominika, Dickinson, Amber, Foster, Andy +133 more·2023·Journal of Neuroinflammation
DOI 10.1186/s12974-023-02769-yPMC PMC10084680
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Abstract

Neuroinflammation is an important component of many neurodegenerative diseases, whether as a primary cause or a secondary outcome. For that reason, either as diagnostic tools or to monitor progression and/or pharmacological interventions, there is a need for robust biomarkers of neuroinflammation in the brain. Mitochondrial TSPO (18 kDa Translocator protein) is one of few available biomarkers of neuroinflammation for which there are clinically available PET imaging agents. In this study, we further characterised neuroinflammation in a mouse model of prion-induced chronic neurodegeneration (ME7) including a pharmacological intervention via a CSF1R inhibitor. This was achieved by autoradiographic binding of the second-generation TSPO tracer, [3H]PBR28, along with a more comprehensive examination of the cellular contributors to the TSPO signal changes by immunohistochemistry. We observed regional increases of TSPO in the ME7 mouse brains, particularly in the hippocampus, cortex and thalamus. This increased TSPO signal was detected in the cells of microglia/macrophage lineage as well as in astrocytes, endothelial cells and neurons. Importantly, we show that the selective CSF1R inhibitor, JNJ-40346527 (JNJ527), attenuated the disease-dependent increase in TSPO signal, particularly in the dentate gyrus of the hippocampus, where JNJ527 attenuated the number of Iba1+ microglia and neurons, but not GFAP+ astrocytes or endothelial cells. These findings suggest that [3H]PBR28 quantitative autoradiography in combination with immunohistochemistry are important translational tools for detecting and quantifying neuroinflammation, and its treatments, in neurodegenerative disease. Furthermore, we demonstrate that although TSPO overexpression in the ME7 brains was driven by various cell types, the therapeutic effect of the CSF1R inhibitor was primarily to modulate TSPO expression in microglia and neurons, which identifies an important route of biological action of this particular CSF1R inhibitor and provides an example of a cell-specific effect of this type of therapeutic agent on the neuroinflammatory process.

AI snapshot

Written from the paper’s full text.

TL;DR

In ME7 prion-infected mice, [3H]PBR28 autoradiography detected increased TSPO binding in cortex, hippocampus, and thalamus, which was attenuated by the CSF1R inhibitor JNJ-40346527. TSPO overexpression arose from microglia/macrophages, astrocytes, neurons, and endothelial cells, but JNJ527 reduced TSPO primarily in Iba1+ microglia/macrophages and neurons in the dentate gyrus.

Key findings

  • [3H]PBR28 binding was significantly increased in ME7 vs NBH mouse brain: mixed-effects ANOVA showed a treatment effect F(2,20)=40.14, p<0.0001, and ROI effect F(1.998,39.96)=11.90, p<0.0001; JNJ527 attenuated binding across regions.
  • [3H]PBR28 showed larger relative ME7 increases than [3H]PK11195 in cortex (~twofold vs 0.9-fold), hippocampus (~1.5-fold vs 0.95-fold), and thalamus (~threefold vs 1.5-fold).
  • TSPO immunohistochemistry was increased in ME7 vs NBH in cortex (p=0.0033), thalamus (p<0.0001), CA1 (p<0.0001), CA3 (p=0.0051), and dentate gyrus (p<0.0001); JNJ527 significantly reduced TSPO only in dentate gyrus (p=0.0090), with a trend in CA1 (p=0.09).
  • In dentate gyrus, ME7 increased TSPO+Iba1+ cells (~2.2-fold, p<0.0001), TSPO+GFAP+ cells (p=0.0012), and TSPO+NeuN+ cells (~1.8-fold, p<0.0001), but not TSPO+CD31+ area (p=0.1571); JNJ527 reduced TSPO+Iba1+ cells (p=0.0341) and TSPO+NeuN+ cells (p<0.0001), but not TSPO+GFAP+ cells (p=0.9648) or TSPO+CD31+ area.

Methods

  • C57BL/6J mice (n=7–8/group) received bilateral CA1 hippocampal injections of ME7 prion brain homogenate or normal brain homogenate at 10 weeks; JNJ-40346527 was given in chow at 30 mg/kg for 4 weeks starting 12 weeks after infection, with tissue collected at 26 weeks.
  • Ex vivo [3H]PBR28 autoradiography (1 nM; non-specific binding with unlabelled PK11195) quantified cortex, hippocampus, and thalamus; TSPO immunohistochemistry and triple immunofluorescence for TSPO with Iba1, GFAP, NeuN, and CD31 assessed regional and cellular TSPO sources, focusing on dentate gyrus.
  • Autoradiography used mixed-effects ANOVA with ROI as within-subject and treatment as between-subject factors plus Sidak post hoc tests; other data used one-way ANOVA with Tukey post hoc tests.

Limitations

  • Stated: DAB-based TSPO immunohistochemistry is non-linear and enzymatically amplified, producing greater variability and lower sensitivity than autoradiography; the authors note this likely explains weaker JNJ527 effects outside dentate gyrus.
  • Stated: the study did not directly measure neurogenesis, so the neuronal findings are interpreted indirectly.
  • Evident: cellular co-localization and cell-count analyses were restricted to the dentate gyrus, and the sample size was small (n=7–8/group), limiting generalization to other regions and human TSPO PET.

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

Integrity signals

Registry

Crossref · 10.1186/s12974-023-02769-y

Cites

63 of 63 references verified0 citation errors · 0 not checkable

Statistics

28 tests recomputed · 2 with the reported and recomputed p on opposite sides of .05

Claims the paper makes

50 of 76 cited sources checked · 23 supported · 4 flagged · accuracy not yet measured

  • Premisecitation

    TSPO is expressed in almost all CNS cells, including astrocytes, neurons and endothelial cells.

    “Although increased TSPO expression has historically been mostly associated with microglial activation, TSPO is now known to be expressed in almost all CNS cells including astrocytes, neurons and endothelial cells [21–27], all of which may also be involved in, or be affected by neuroinflammation [28].”
    Quote in the paperSubstantial claim5 of 7 sources checked
    • cites [21] Vicente-Rodríguez M, Singh N, Turkheimer F, Peris-Yague A, Randall K, Veronese M, et al. Resolving the cellular specificity of TSPO imaging in a rat model of peripherally-induced neuroinflammation. Brain Behav Immun. 2021;96:154–67. source

      The Results section confirms TSPO expression in astrocytes, neurons and endothelial cells in the rat brain regions examined; the Introduction also describes TSPO in various other brain cells. The paper does not establish the broader claim that it is expressed in almost all CNS cells. Page numbers were not available in our copy. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [22] Nutma E, Ceyzériat K, Amor S, Tsartsalis S, Millet P, Owen DR, et al. Cellular sources of TSPO expression in healthy and diseased brain. Eur J Nucl Med Mol Imaging. 2021;49:146–63. source

      The Introduction and Conclusion say TSPO occurs in astrocytes and endothelial cells as well as microglia. But the Wild-type section says its expression varies by brain region and that neurons generally lack TSPO in the mouse brain, while noting particular neuronal exceptions. The source therefore does not support the unqualified claim that TSPO is expressed in almost all CNS cells. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [23] Girard C, Liu S, Adams D, Lacroix C, Sinéus M, Boucher C, et al. Axonal regeneration and neuroinflammation: roles for the translocator protein 18 kDa. J Neuroendocrinol. 2012;24:71–81. source
    • cites [24] Mills CD, Bitler JL, Woolf CJ. Role of the peripheral benzodiazepine receptor in sensory neuron regeneration. Mol Cell Neurosci. 2005;30:228–37. source
    • cites [25] Betlazar C, Harrison-Brown M, Middleton RJ, Banati R, Liu GJ. Cellular sources and regional variations in the expression of the neuroinflammatory marker translocator protein (TSPO) in the normal brain. Int J Mol Sci. 2018;19(9):2707. source

      Results §2.2 identifies vascular endothelial cells as a major source of TSPO in the normal mouse brain, but reports no discernible staining in astrocytes or mature NeuN+ neurons across the brain. It does find expression in particular neuronal populations, including Purkinje cells; that does not support the claim that TSPO is expressed in almost all CNS cells. Page numbers were not supplied in our copy. (LLM, conf=0.88)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [26] Pannell M, Economopoulos V, Wilson TC, Kersemans V, Isenegger PG, Larkin JR, et al. Imaging of translocator protein upregulation is selective for pro-inflammatory polarized astrocytes and microglia. Glia. 2020;68:280–97. source

      The Introduction says that, in the CNS, TSPO expression is confined to glial and ependymal cells, contrary to the claim that it is expressed in almost all CNS cells, including neurons. The Discussion does report endothelial TSPO expression, and the paper documents astrocytic expression, but neither supports the claim about neurons or nearly all CNS cells. Page numbers were not supplied in our copy. (LLM, conf=0.86)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [27] Gui Y, Marks JD, Das S, Hyman BT, Serrano-Pozo A. Characterization of the 18 kDa translocator protein (TSPO) expression in post-mortem normal and Alzheimer’s disease brains. Brain Pathol. 2020;30:151–64. source

      The Results section confirms TSPO expression in astrocytes and endothelial cells, but reports that neuronal-marker staining showed no TSPO expression in neurons. That directly contradicts the claim’s inclusion of neurons among TSPO-expressing CNS cells. (LLM, conf=0.90)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
  • Premisecitation

    CSF1R is thought to be expressed exclusively by microglia.

    “As CSF1R is thought to be expressed exclusively by microglia [34, 63, 64]”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [34] Gomez-Nicola D, Fransen NL, Suzzi S, Perry VH. Regulation of microglial proliferation during chronic neurodegeneration. J Neurosci. 2013;33:2481–93. source

      The Discussion (page unavailable) describes CSF1R blockade in microglia and says the same approach had previously controlled expansion of other macrophage populations. That acknowledgment runs contrary to the unqualified claim that CSF1R is expressed exclusively by microglia. (LLM, conf=0.78)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [63] Erblich B, Zhu L, Etgen AM, Dobrenis K, Pollard JW. Absence of colony stimulation factor-1 receptor results in loss of microglia, disrupted brain development and olfactory deficits. PLoS ONE. 2011;6: e26317. source

      The Introduction says CSF-1R is expressed solely on microglia postnatally, and the Discussion limits the finding to the normal brain regions and stages examined (page numbers unavailable). The unqualified claim is broader: the paper also describes CSF-1R in macrophages and reports neuronal expression after brain injury. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [64] Sasmono RT, Oceandy D, Pollard JW, Tong W, Pavli P, Wainwright BJ, et al. A macrophage colony-stimulating factor receptor–green fluorescent protein transgene is expressed throughout the mononuclear phagocyte system of the mouse. Blood. 2003;101:1155–63. source
  • Premisecitation

    The selective CSF1R inhibitor JNJ527 significantly blocked microglial proliferation in the ME7 mouse model of prion disease, as detected by ex vivo autoradiography with [3H]PK11195.

    “We previously demonstrated that the selective CSF1R inhibitor JNJ527 significantly blocked microglial proliferation in the ME7 mouse model of prion disease, and this has been detected by ex vivo autoradiography with the first generation TSPO ligand, [3H]PK11195 [33].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [33] Mancuso R, Fryatt G, Cleal M, Obst J, Pipi E, Monzón-Sandoval J, et al. CSF1R inhibitor JNJ-40346527 attenuates microglial proliferation and neurodegeneration in P301S mice. Brain. 2019;142:3243–64. source
  • Premisecitation

    In the ME7 mouse, the selective CSF1R inhibitor JNJ527 significantly blocks microglial proliferation, as shown by reduced [3H]PK11195 binding on autoradiography.

    “Using the ME7 mouse, we previously corroborated the ability of the selective colony stimulating factor receptor 1 (CSF1R) inhibitor, JNJ-40346527 (JNJ527), to significantly block microglial proliferation. This was demonstrated by the reduced binding of the first generation TSPO tracer, [3H]PK11195, using autoradiography [33].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [33] Mancuso R, Fryatt G, Cleal M, Obst J, Pipi E, Monzón-Sandoval J, et al. CSF1R inhibitor JNJ-40346527 attenuates microglial proliferation and neurodegeneration in P301S mice. Brain. 2019;142:3243–64. source
  • Premisecitation

    Clinical use of second-generation TSPO tracers is complicated by a single nucleotide polymorphism in the Tspo gene that alters binding affinity in a proportion of humans.

    “However, the clinical use of these second-generation tracers is complicated by a single nucleotide polymorphism in the Tspo gene that alters its binding affinity in a proportion of humans.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [19] Owen DR, Yeo AJ, Gunn RN, Song K, Wadsworth G, Lewis A, et al. An 18-kDa translocator protein (TSPO) polymorphism explains differences in binding affinity of the PET radioligand PBR28. J Cereb Blood Flow Metab. 2012;32:1–5. source
    • cites [20] Mizrahi R, Rusjan PM, Kennedy J, Pollock B, Mulsant B, Suridjan I, et al. Translocator protein (18 kDa) polymorphism (rs6971) explains in-vivo brain binding affinity of the PET radioligand [18 F]-FEPPA. J Cereb Blood Flow Metab. 2012;32:968–72. source
  • Classified as probably not a substantial claim. 32
  • May combine two claims. 9

  • Classified as probably not a substantial claim.

    Premisecitation

    Chronic neuroinflammation mediated by disease-associated activation of microglia and astrocytes is a common pathological hallmark of neurodegeneration.

    “Another common pathological hallmark of neurodegeneration appears to be chronic neuroinflammation, mediated by disease-associated activation of microglia and astrocytes [1].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [1] Kwon HS, Koh S-H. Neuroinflammation in neurodegenerative disorders: the roles of microglia and astrocytes. Transl Neurodegener. 2020;9:42. source

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

    Premisecitation

    PET imaging has shown increased TSPO signal in a range of human neurodegenerative diseases with an inflammatory component.

    “Accordingly, using PET imaging, increased TSPO signal has been observed in a range of human neurodegenerative diseases with an inflammatory component, such as simian immunodeficiency virus encephalitis [10], multiple sclerosis [11], Huntington’s [12] and Alzheimer’s disease [13].”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites [10] Vera JH, Guo Q, Cole JH, Boasso A, Greathead L, Kelleher P, et al. Neuroinflammation in treated HIV-positive individuals: a TSPO PET study. Neurology. 2016;86:1425–32. source

      The Discussion (page unavailable) reports increased brain TSPO radioligand uptake on PET in treated HIV-positive humans. That supports an example of increased TSPO signal, but this paper does not establish the claimed range of diseases or the sentence’s specifically cited example of simian immunodeficiency virus encephalitis. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [11] Banati RB, Newcombe J, Gunn RN, Cagnin A, Turkheimer F, Heppner F, et al. The peripheral benzodiazepine binding site in the brain in multiple sclerosis. Brain. 2000;123:2321–37. source
    • cites [12] Meßmer K, Reynolds GP. Increased peripheral benzodiazepine binding sites in the brain of patients with Huntington’s disease. Neurosci Lett. 1998;241:53–6. source

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

    Premisecitation

    [11C]PK11195 is a TSPO PET tracer still routinely used in clinical imaging but has a poor signal-to-noise ratio due to low binding affinity.

    “One of the earliest TSPO PET tracers is [11C]PK11195, still routinely used in clinical imaging but hampered by the poor signal-to-noise ratio arising from its low binding affinity [16, 17].”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [16] Turkheimer FE, Rizzo G, Bloomfield PS, Howes O, Zanotti-Fregonara P, Bertoldo A, et al. The methodology of TSPO imaging with positron emission tomography. Biochem Soc Trans. 2015;43:586–92. source

      The section on [11C]-(R)-PK11195 calls it the radiotracer of choice for imaging neuroinflammation in vivo. The second-generation radioligands section says newer tracers were developed to improve its signal-to-noise properties, but does not establish that low binding affinity causes the poorer signal-to-noise ratio; it describes PK11195 as having nanomolar affinity. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [17] Cumming P, Burgher B, Patkar O, Breakspear M, Vasdev N, Thomas P, et al. Sifting through the surfeit of neuroinflammation tracers. J Cereb Blood Flow Metab. 2018;38:204–24. source

      The Introduction identifies [11C]-(R)-PK11195 as a prototype TSPO PET tracer used in patients and reports its low specific imaging signal. It does not attribute that signal specifically to low binding affinity; instead, it suggests low brain free fraction and/or competition from endogenous ligands. The Conclusions also say [11C]-labelled tracers are not widely available for routine use, so routine clinical use is not established here. Page numbers are unavailable in our copy. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

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

    Premisecitation

    JNJ527 attenuates microglia proliferation in models of neurodegeneration.

    “CSF1R inhibitor JNJ527 is a tool compound that was previously shown to attenuate microglia proliferation and provide therapeutic action in models of neurodegeneration including ME7 prion mice and in the P301S mouse tauopathy model [33].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [33] Mancuso R, Fryatt G, Cleal M, Obst J, Pipi E, Monzón-Sandoval J, et al. CSF1R inhibitor JNJ-40346527 attenuates microglial proliferation and neurodegeneration in P301S mice. Brain. 2019;142:3243–64. source

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

    Premisecitation

    JNJ527 provides therapeutic action in models of neurodegeneration including ME7 prion mice and P301S tauopathy mice.

    “CSF1R inhibitor JNJ527 is a tool compound that was previously shown to attenuate microglia proliferation and provide therapeutic action in models of neurodegeneration including ME7 prion mice and in the P301S mouse tauopathy model [33].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [33] Mancuso R, Fryatt G, Cleal M, Obst J, Pipi E, Monzón-Sandoval J, et al. CSF1R inhibitor JNJ-40346527 attenuates microglial proliferation and neurodegeneration in P301S mice. Brain. 2019;142:3243–64. source

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

    Premisecitation

    Astrocytes, neurons and endothelial cells may also be involved in or affected by neuroinflammation.

    “all of which may also be involved in, or be affected by neuroinflammation [28].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [28] Carson MJ, Thrash JC, Walter B. The cellular response in neuroinflammation: the role of leukocytes, microglia and astrocytes in neuronal death and survival. Clin Neurosci Res. 2006;6:237–45. source

  • Classified as probably not a substantial claim.

    Premisecitation

    Prion disease experimental models are robust and tractable laboratory tools for studying these pathological processes.

    “Prion disease experimental models are robust and tractable laboratory tools to study these pathological processes [31, 32].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [31] Gomez-Nicola D, Suzzi S, Vargas-Caballero M, Fransen NL, Al-Malki H, Cebrian-Silla A, et al. Temporal dynamics of hippocampal neurogenesis in chronic neurodegeneration. Brain. 2014;137:2312–28. source
    • cites [32] Gomez-Nicola D, Perry VH. Microglial dynamics and role in the healthy and diseased brain. Neurosci. 2015;21:169–84. source

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

    Premisecitation

    In postmortem human Alzheimer's disease brains, TSPO positive microglia, astrocytes and endothelium are all increased.

    “Similarly, others have found that TSPO positive microglia, astrocytes and endothelium are all increased in models of Alzheimer’s-like neurodegeneration [48–51] and in postmortem human Alzheimer’s disease brains [27].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [27] Gui Y, Marks JD, Das S, Hyman BT, Serrano-Pozo A. Characterization of the 18 kDa translocator protein (TSPO) expression in post-mortem normal and Alzheimer’s disease brains. Brain Pathol. 2020;30:151–64. source

      The Results section identifies TSPO-positive microglia, astrocytes, and endothelial cells in human brain tissue. It reports a modest increase in overall TSPO immunoreactivity in AD white matter, but does not show that TSPO-positive cells of each of the three types increased; the Conclusions instead emphasize substantial overlap in TSPO levels between AD and controls. Page numbers were not available in our copy. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    Aberrant increased neurogenesis takes place during some neurodegenerative sequelae, including experimental prion disease.

    “Our interest in the dentate gyrus was furthered by earlier discoveries that aberrant increased neurogenesis takes place during some neurodegenerative sequelae including that in experimental prion disease [31].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [31] Gomez-Nicola D, Suzzi S, Vargas-Caballero M, Fransen NL, Al-Malki H, Cebrian-Silla A, et al. Temporal dynamics of hippocampal neurogenesis in chronic neurodegeneration. Brain. 2014;137:2312–28. source

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

    Premisecitation

    Gliosis is consistent with the presence of neuroinflammation.

    “Gliosis is consistent with the presence of neuroinflammation [46].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [46] Bernaus A, Blanco S, Sevilla A. Glia crosstalk in neuroinflammatory diseases. Front Cell Neurosci. 2020;14:1–17. source

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

    Premisecitation

    The dentate gyrus is one of the two principal neurogenic niches, where stem cells can differentiate into neuroblasts that have the potential to migrate to sites of injury in a potential compensatory neural repair role.

    “The dentate gyrus is also one of the two principal neurogenic niches, where stem cells can differentiate into neuroblasts that have the potential to migrate to sites of injury in a potential compensatory neural repair role [42, 43].”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [42] Keller D, Erö C, Markram H. Cell densities in the mouse brain: a systematic review. Front Neuroanat. 2018;12:83. source

      The Results section, “Hippocampus and associated areas” (page unavailable), identifies the dentate gyrus and reports cell densities, but does not describe it as a neurogenic niche. Searches for stem-cell differentiation, neuroblast migration, injury, and neural repair found no passage in our copy addressing those claims. (LLM, conf=0.73)

      Experimental — accuracy not yet measured
    • cites [43] Kesner RP. An analysis of the dentate gyrus function. Behav Brain Res. 2013;254:1–7. source

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

    Premisecitation

    Neurosteroids are thought to be associated with promoting adult neurogenesis.

    “Neurosteroids are also thought to be associated with promoting adult neurogenesis, alongside their other proposed roles of modulating synaptic transmission and neuronal network formation [58].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [58] Tozzi A, Bellingacci L, Pettorossi VE. Rapid estrogenic and androgenic neurosteroids effects in the induction of long-term synaptic changes: implication for early memory formation. Front Neurosci. 2020;14: 572511. source

      The Introduction (page unavailable) says sex neurosteroids influence neural-circuit development and help direct synaptogenesis and changes in synaptic transmission. It does not say they promote adult neurogenesis, which is distinct from forming synapses or dendritic spines; the term appears in the title of a work listed in the References, not as a finding stated by this paper. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

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

    Premisecitation

    In a previous study of LPS-challenged rats (a rodent model of acute neuroinflammation), TSPO signal was present in all examined cells at baseline.

    “our previous study examining cellular sources of TSPO in a rodent model of acute neuroinflammation, the LPS-challenged rat, also found a similar diversity of source of TSPO signal which was present in all examined cells at baseline, but then increased significantly in astrocytes and microglia upon inflammatory insult [21].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [21] Vicente-Rodríguez M, Singh N, Turkheimer F, Peris-Yague A, Randall K, Veronese M, et al. Resolving the cellular specificity of TSPO imaging in a rat model of peripherally-induced neuroinflammation. Brain Behav Immun. 2021;96:154–67. source

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

    Premisecitation

    In models of Alzheimer's-like neurodegeneration, TSPO positive microglia, astrocytes and endothelium are all increased.

    “Similarly, others have found that TSPO positive microglia, astrocytes and endothelium are all increased in models of Alzheimer’s-like neurodegeneration [48–51] and in postmortem human Alzheimer’s disease brains [27].”
    Quote in the paperNot a substantial claim4 of 4 sources checked
    • cites [48] Ji B, Maeda J, Sawada M, Ono M, Okauchi T, Inaji M, et al. Imaging of peripheral benzodiazepine receptor expression as biomarkers of detrimental versus beneficial glial responses in mouse models of Alzheimer’s and other CNS pathologies. J Neurosci. 2008;28:12255–67. source

      The Results section, “Distinct cellular localizations of PBR in models of Aβ and tau pathologies” (page unavailable), reports PBR-positive astrocytes near plaques in APP23 mice and PBR-positive microglia in PS19 tau mice. It does not report increased PBR-positive endothelium. It also distinguishes the two models rather than finding that all three cell types increased in each. (LLM, conf=0.89)

      Experimental — accuracy not yet measured
    • cites [49] Liu B, Le KX, Park M-A, Wang S, Belanger AP, Dubey S, et al. In vivo detection of age- and disease-related increases in neuroinflammation by 18 F-GE180 TSPO MicroPET imaging in wild-type and Alzheimer’s transgenic mice. J Neurosci. 2015;35:15716–30. source

      The colocalization results (page not available) show TSPO-positive microglia associated with amyloid plaques in APP/PS1dE9 mice. They report only rare overlap between TSPO and the astrocyte marker GFAP, and the passages examined do not establish an increase in TSPO-positive endothelium. The source therefore does not support the claim that all three cell types are increased. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [50] Tournier BB, Tsartsalis S, Rigaud D, Fossey C, Cailly T, Fabis F, et al. TSPO and amyloid deposits in sub-regions of the hippocampus in the 3xTgAD mouse model of Alzheimer’s disease. Neurobiol Dis. 2019;121:95–105. source

      The Discussion (p. 20) reports elevated TSPO colocalization with microglia in 3xTgAD mice, with some TSPO expression in astrocytes. It does not establish that TSPO-positive astrocytes increased. The endothelial finding (pp. 17, 21) concerns TSPO-positive cells in human choroid plexus, not an increase in an Alzheimer’s-like mouse model. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [51] Cosenza-Nashat M, Zhao ML, Suh HS, Morgan J, Natividad R, Morgello S, et al. Expression of the translocator protein of 18 kDa by microglia, macrophages and astrocytes based on immunohistochemical localization in abnormal human brain. Neuropathol Appl Neurobiol. 2009;35(3):306–28. source

      The Results section, “Expression of TSPO in AD” (page unavailable), reports widespread TSPO-positive glia in human AD brains, predominantly microglia, with some astrocytes. It also notes TSPO-positive vessels, but does not show that endothelial TSPO increased. The Discussion (page unavailable) describes endothelial TSPO as potentially constitutive and reports a decrease in vascular staining with AD. This citation attaches to the human postmortem AD portion of the sentence, not the animal-model portion. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    TSPO is involved in the mitochondrial intermembrane transport of cholesterol.

    “TSPO is involved in the mitochondrial intermembrane transport of cholesterol”
    Quote in the paperSubstantial claim1 of 4 sources checked
    • cites [53] Ferzaz B, Brault E, Bourliaud G, Robert J-P, Poughon G, Claustre Y, et al. SSR180575 (7-chloro-N, N,5-trimethyl-4-oxo-3-phenyl-3,5-dihydro-4H-pyridazino[4,5-b]indole-1-acetamide), a peripheral benzodiazepine receptor ligand, promotes neuronal survival and repair. J Pharmacol Exp Ther. 2002;301:1067–78. source
    • cites [54] Korneyev A, Pan BS, Polo A, Romeo E, Guidotti A, Costa E. Stimulation of brain pregnenolone synthesis by mitochondrial diazepam binding inhibitor receptor ligands in vivo. J Neurochem. 1993;61:1515–24. source
    • cites [55] Lacor P, Gandolfo P, Tonon M-C, Brault E, Dalibert I, Schumacher M, et al. Regulation of the expression of peripheral benzodiazepine receptors and their endogenous ligands during rat sciatic nerve degeneration and regeneration: a role for PBR in neurosteroidogenesis. Brain Res. 1999;815:70–80. source
    • cites [56] Papadopoulos V, Amri H, Boujrad N, Cascio C, Culty M, Garnier M, et al. Peripheral benzodiazepine receptor in cholesterol transport and steroidogenesis. Steroids. 1997;62:21–8. source

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

    Premisecitation

    Microgliosis and astrocytosis were previously described in the dentate gyrus area of the hippocampus of prion disease mice.

    “Microgliosis and astrocytosis were previously described in the dentate gyrus area of the hippocampus of prion disease mice [32, 38, 39].”
    Quote in the paperNot a substantial claim3 of 3 sources checked
    • cites [32] Gomez-Nicola D, Perry VH. Microglial dynamics and role in the healthy and diseased brain. Neurosci. 2015;21:169–84. source

      The section “Where Do Microglial Cells Come From? Migrating and Proliferating Microglia” reports a tenfold expansion of the microglial population in experimental prion disease, supporting the microgliosis component. It does not place that finding in the dentate gyrus or report astrocytosis there in prion-disease mice; searches for those terms found no such passage. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [38] De Lucia C, Rinchon A, Olmos-Alonso A, Riecken K, Fehse B, Boche D, et al. Microglia regulate hippocampal neurogenesis during chronic neurodegeneration. Brain Behav Immun. 2016;55:179–90. source

      The Results section reports significantly increased microglial density across the dentate gyrus in ME7 prion-diseased mice, supporting microgliosis there. The passages from our copy do not report astrocytosis in that area, so they support only part of the cited sentence. No page number is available in our copy. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [39] Asuni AA, Gray B, Bailey J, Skipp P, Perry VH, O’Connor V. Analysis of the hippocampal proteome in ME7 prion disease reveals a predominant astrocytic signature and highlights the brain-restricted production of clusterin in chronic neurodegeneration. J Biol Chem. 2014;289:4532–1545. source

      The Results (Fig. 3; page unavailable) show reactive astrocytes in the hippocampus of ME7 prion-disease mice and label the dentate gyrus. The Results also report microglial proliferation during prion disease, but our copy does not establish that microgliosis occurred specifically in the dentate gyrus. (LLM, conf=0.83)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    Cholesterol is the necessary precursor for the synthesis of neurosteroids.

    “cholesterol which is the necessary precursor for the synthesis of neurosteroids [53–56]”
    Quote in the paperSubstantial claim1 of 4 sources checked
    • cites [53] Ferzaz B, Brault E, Bourliaud G, Robert J-P, Poughon G, Claustre Y, et al. SSR180575 (7-chloro-N, N,5-trimethyl-4-oxo-3-phenyl-3,5-dihydro-4H-pyridazino[4,5-b]indole-1-acetamide), a peripheral benzodiazepine receptor ligand, promotes neuronal survival and repair. J Pharmacol Exp Ther. 2002;301:1067–78. source
    • cites [54] Korneyev A, Pan BS, Polo A, Romeo E, Guidotti A, Costa E. Stimulation of brain pregnenolone synthesis by mitochondrial diazepam binding inhibitor receptor ligands in vivo. J Neurochem. 1993;61:1515–24. source
    • cites [55] Lacor P, Gandolfo P, Tonon M-C, Brault E, Dalibert I, Schumacher M, et al. Regulation of the expression of peripheral benzodiazepine receptors and their endogenous ligands during rat sciatic nerve degeneration and regeneration: a role for PBR in neurosteroidogenesis. Brain Res. 1999;815:70–80. source
    • cites [56] Papadopoulos V, Amri H, Boujrad N, Cascio C, Culty M, Garnier M, et al. Peripheral benzodiazepine receptor in cholesterol transport and steroidogenesis. Steroids. 1997;62:21–8. source

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

    Premisecitation

    In the LPS-challenged rat model, TSPO signal increased significantly in astrocytes and microglia upon inflammatory insult.

    “our previous study examining cellular sources of TSPO in a rodent model of acute neuroinflammation, the LPS-challenged rat, also found a similar diversity of source of TSPO signal which was present in all examined cells at baseline, but then increased significantly in astrocytes and microglia upon inflammatory insult [21].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [21] Vicente-Rodríguez M, Singh N, Turkheimer F, Peris-Yague A, Randall K, Veronese M, et al. Resolving the cellular specificity of TSPO imaging in a rat model of peripherally-induced neuroinflammation. Brain Behav Immun. 2021;96:154–67. source

  • Classified as probably not a substantial claim.

    Premisecitation

    TSPO is expressed in glial and endothelial cells.

    “along with the established presence in the glial and endothelial cells [26, 45]”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [26] Pannell M, Economopoulos V, Wilson TC, Kersemans V, Isenegger PG, Larkin JR, et al. Imaging of translocator protein upregulation is selective for pro-inflammatory polarized astrocytes and microglia. Glia. 2020;68:280–97. source
    • cites [45] Betlazar C, Harrison-Brown M, Middleton RJ, Banati R, Liu GJ. Cellular sources and regional variations in the expression of the neuroinflammatory marker translocator protein (TSPO) in the normal brain. Int J Mol Sci. 2018;19:2707. source

  • Classified as probably not a substantial claim.

    Premisecitation

    An increase in hippocampal neurogenesis correlates with expansion of the microglia population.

    “an increase in hippocampal neurogenesis was previously shown to correlate with the expansion of the microglia population [31]”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [31] Gomez-Nicola D, Suzzi S, Vargas-Caballero M, Fransen NL, Al-Malki H, Cebrian-Silla A, et al. Temporal dynamics of hippocampal neurogenesis in chronic neurodegeneration. Brain. 2014;137:2312–28. source

      The Results report increased hippocampal neurogenesis in prion-diseased mice, and the Discussion describes expansion of their microglial population. But the Discussion says the two responses *could* be related and that further analysis is needed; it does not show the correlation claimed. (LLM, conf=0.84)

      Experimental — accuracy not yet measured

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

    Premisecitation

    GW2580 restores normal neuronal differentiation.

    “pharmacological modulation of microglia with another CSF1R inhibitor, GW2580 was able to reduce neuronal population and aberrant neurogenesis and restore the normal neuronal differentiation [38].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [38] De Lucia C, Rinchon A, Olmos-Alonso A, Riecken K, Fehse B, Boche D, et al. Microglia regulate hippocampal neurogenesis during chronic neurodegeneration. Brain Behav Immun. 2016;55:179–90. source

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

    Premisecitation

    GW2580 reduces aberrant neurogenesis.

    “pharmacological modulation of microglia with another CSF1R inhibitor, GW2580 was able to reduce neuronal population and aberrant neurogenesis and restore the normal neuronal differentiation [38].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [38] De Lucia C, Rinchon A, Olmos-Alonso A, Riecken K, Fehse B, Boche D, et al. Microglia regulate hippocampal neurogenesis during chronic neurodegeneration. Brain Behav Immun. 2016;55:179–90. source

  • May combine two claims.

    Interpretationcitation

    The reduction of Iba1+ microglia/macrophages by JNJ527 is unsurprising because CSF1R is expressed by microglia and its inhibition reduces microglial proliferation.

    “As CSF1R is thought to be expressed exclusively by microglia [34, 63, 64] and its inhibition reduces their proliferation, it is unsurprising that Iba1+ microglia/macrophages were also reduced by this treatment.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [34] Gomez-Nicola D, Fransen NL, Suzzi S, Perry VH. Regulation of microglial proliferation during chronic neurodegeneration. J Neurosci. 2013;33:2481–93. source

      In “Signaling through CSF1R controls microglial proliferation” (page unavailable), the paper reports CSF1R on microglia and finds that blocking it reduces their proliferation and population expansion in mouse hippocampus. The cited passages do not establish that CSF1R is expressed exclusively by microglia. The paper tests a blocking antibody and GW2580, not JNJ527; the sentence’s observation about JNJ527 is the citing authors’ own inference. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [63] Erblich B, Zhu L, Etgen AM, Dobrenis K, Pollard JW. Absence of colony stimulation factor-1 receptor results in loss of microglia, disrupted brain development and olfactory deficits. PLoS ONE. 2011;6: e26317. source
    • cites [64] Sasmono RT, Oceandy D, Pollard JW, Tong W, Pavli P, Wainwright BJ, et al. A macrophage colony-stimulating factor receptor–green fluorescent protein transgene is expressed throughout the mononuclear phagocyte system of the mouse. Blood. 2003;101:1155–63. source
    • Rests on an assumption: Inhibition of CSF1R reduces microglial proliferation.

  • May combine two claims.

    Findingown data

    [3H]PBR28 showed a larger relative increase in binding to ME7 brains compared to NBH controls than [3H]PK11195 in cortex (~twofold vs. 0.9-fold), hippocampus (~1.5-fold vs. 0.95-fold) and thalamus (~threefold vs. 1.5-fold).

    “and a larger relative increase in binding to the ME7 brains compared to NBH controls with [3H]PBR28 compared to [3H]PK11195 in cortex (~ twofold vs. 0.9-fold), hippocampus (~ 1.5-fold vs. 0.95-fold) and thalamus (~ threefold vs. 1.5-fold) (Additional file 1: Fig. S1).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Mixed-effects model ANOVA of [3H]PBR28 autoradiography showed a significant effect of treatment group (p < 0.0001) and ROI (p < 0.0001) with no significant group × ROI interaction (p = 0.0833).

    “Mixed-effects model ANOVA analysis of [3H]PBR28 autoradiography showed a significant effect of treatment group (p < 0.0001) and ROI (p < 0.0001) with no significant group × ROI interaction (p = 0.0833) (see Table 1A).”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Second-generation TSPO PET tracers with increased affinity, including [11C]PBR28, have been developed.

    “To overcome this problem, numerous ‘second-generation’ TSPO PET tracers with increased affinity have been developed, including [11C]PBR28 [18].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [18] Alam MM, Lee J, Lee S-Y. Recent progress in the development of TSPO PET ligands for neuroinflammation imaging in neurological diseases. Nucl Med Mol Imaging. 2010;2017(51):283–96.

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Study design and conductnone given

    Slides were exposed to tritium-sensitive film together with a set of tritium standards for calibration.

    “Slides were exposed to tritium-sensitive film (Amersham Hyperfilm MP, GE Healthcare) in autoradiography cassettes together with a set of tritium standards ([3H] Microscale, American Radiolabelled Chemicals, #art-0123A) for 6 weeks.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Prion diseases are chronic transmissible neurodegenerative diseases characterized by progressive neuronal loss and neuroinflammation.

    “Prion diseases are a group of chronic transmissible neurodegenerative diseases, characterized by both progressive neuronal loss and neuroinflammation [29, 30].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [29] Masters CL, Richardson EP. Subacute spongiform encephalopathy (Creutzfeldt-Jakob disease). Brain. 1978;101:333–44. source
    • cites [30] Williams A, Lucassen PJ, Ritchie D, Bruce M. PrP deposition, microglial activation, and neuronal apoptosis in murine scrapie. Exp Neurol. 1997;144:433–8. source

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Non-specific binding was determined on adjacent sections in the presence of unlabelled PK11195 (20 μM).

    “Non-specific binding was determined on adjacent sections in the presence of unlabelled PK11195 (20 µM; Sigma-Aldrich, #85532-75-8).”
    Quote in the paperSubstantial claim

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

    Interpretationreasoning

    If there are more neurons and neurons contain TSPO, then TSPO signal is expected to increase.

    “if there are more neurons, and neurons contain TSPO, then it is expected that TSPO signal will increase.”
    Quote in the paperSubstantial claim

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

    Interpretationcitation

    The observed increase in neurons potentially reflects the aberrant placement of neurons.

    “and potentially reflects the aberrant placement of neurons [47]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [47] Winner B, Kohl Z, Gage FH. Neurodegenerative disease and adult neurogenesis. Eur J Neurosci. 2011;33:1139–51. source

  • Classified as probably not a substantial claim.

    Premisecitation

    TSPO is present in differentiating/regenerating neurons.

    “Studies have demonstrated the presence of TSPO in immature neurons, as well as in differentiating/regenerating neurons [23, 24, 52].”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [23] Girard C, Liu S, Adams D, Lacroix C, Sinéus M, Boucher C, et al. Axonal regeneration and neuroinflammation: roles for the translocator protein 18 kDa. J Neuroendocrinol. 2012;24:71–81. source
    • cites [24] Mills CD, Bitler JL, Woolf CJ. Role of the peripheral benzodiazepine receptor in sensory neuron regeneration. Mol Cell Neurosci. 2005;30:228–37. source
    • cites [52] Varga B, Markó K, Hádinger N, Jelitai M, Demeter K, Tihanyi K, et al. Translocator protein (TSPO 18 kDa) is expressed by neural stem and neuronal precursor cells. Neurosci Lett. 2009;462:257–62. source

  • Classified as probably not a substantial claim.

    Premisecitation

    Cholesterol and neurosteroids are important for neuronal regeneration.

    “both cholesterol and neurosteroids are important for neuronal regeneration [57].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [57] Schumacher M, Robel P, Baulieu E-E. Development and regeneration of the nervous system: a role for neurosteroids (part 1 of 2). Dev Neurosci. 1996;18:6–13. source

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

    Study design and conductreasoning

    We did not measure neurogenesis directly in this study.

    “although we did not measure neurogenesis directly here as this was beyond the scope of the study.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    TSPO has a plausible modulatory role in prion neurodegeneration and neurogenesis.

    “All this, therefore, points toward a plausible role for modulatory function of TSPO in prion neurodegeneration and neurogenesis.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    TSPO is present in immature neurons.

    “Studies have demonstrated the presence of TSPO in immature neurons, as well as in differentiating/regenerating neurons [23, 24, 52].”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [23] Girard C, Liu S, Adams D, Lacroix C, Sinéus M, Boucher C, et al. Axonal regeneration and neuroinflammation: roles for the translocator protein 18 kDa. J Neuroendocrinol. 2012;24:71–81. source
    • cites [24] Mills CD, Bitler JL, Woolf CJ. Role of the peripheral benzodiazepine receptor in sensory neuron regeneration. Mol Cell Neurosci. 2005;30:228–37. source
    • cites [52] Varga B, Markó K, Hádinger N, Jelitai M, Demeter K, Tihanyi K, et al. Translocator protein (TSPO 18 kDa) is expressed by neural stem and neuronal precursor cells. Neurosci Lett. 2009;462:257–62. source
36 of 36

  • The paper relies on this without support.

    The TSPO low-affinity binder polymorphism has not been detected in rodents.

    “Interestingly, this phenomenon has not been detected in rodents.”

  • The paper relies on this without support.

    TSPO PET tracers are used to monitor response in clinical trials of novel therapies for neurodegeneration.

    “Imaging neuroinflammation in neurodegeneration with TSPO PET tracers to monitor response in clinical trials of novel therapies has been a major focus of interest in past years.”

  • The paper relies on this without support.

    No clear consensus has been reached regarding the cellular and functional interpretation of the TSPO PET signal.

    “However, no clear consensus has yet been reached regarding the cellular and functional interpretation of the TSPO PET signal.”

  • The paper relies on this without support.

    The lack of consensus on TSPO PET signal interpretation is most likely driven by heterogeneity of TSPO expression in the brain, its cellular sources, and binding affinity differences of TSPO PET imaging agents.

    “This is most likely driven by the heterogeneity of TSPO expression in the brain, its cellular sources, as well as binding affinity differences of TSPO PET imaging agents.”

  • The paper relies on this without support.

    [3H]PBR28 binding reflects TSPO expression.

    “confirmed the increased binding, and, therefore, expression of TSPO”

  • The paper relies on this without support.

    JNJ527 inhibits CSF1R.

    “pharmacological inhibition of CSF1R with JNJ527 attenuated the increased [3H]PBR28 binding”

  • The paper relies on this without support.

    Autoradiography is more representative of the underlying ground truth than immunohistochemistry.

    “our immunohistochemistry measurements are likely to be less sensitive and not as representative of the underlying ground truth, compared to autoradiography.”

  • The paper relies on this without support.

    Inhibition of CSF1R reduces microglial proliferation.

    “and its inhibition reduces their proliferation”

  • The paper relies on this without support.

    The reduction in TSPO-positive microglia/macrophages and neurons after JNJ527 reflects reduced cell numbers rather than reduced TSPO expression per cell.

    “as a consequence of the decrease in the neuronal and Iba1+ microglial population”
9 of 9

Statistics that don’t add up

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

  • Inconsistent p-value across alpha = .05: F (2,18) = 2.606, p = .0003

    Major

    The paper reports p = .0003 here; recomputing from the test statistic and its degrees of freedom gives p = .101. The two fall on opposite sides of alpha = .05. The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .0003 · recomputed p = .101

  • Inconsistent p-value across alpha = .05: F (2,20) = 1.096, p = .0017

    Major

    The paper reports p = .0017 here; recomputing from the test statistic and its degrees of freedom gives p = .353. The two fall on opposite sides of alpha = .05. The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .0017 · recomputed p = .353

  • Inconsistent p-value: F (2,20) = 10.84, p = .0012

    Minor

    The paper reports p = .0012 here; recomputing from the test statistic and its degrees of freedom gives p < .001. The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p = .0012 · recomputed p < .001

  • Inconsistent p-value: F (2,16) = 8.395, p < .0001

    Minor

    The paper reports p < .0001 here; recomputing from the test statistic and its degrees of freedom gives p = .003. The check cannot tell whether the statistic or the p-value is the misprint.

    Reported p < .0001 · recomputed p = .003

4 of 4

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(2, 18) = 2.606reported p = .0003 · recomputed p = .101The two fall on opposite sides of .05

    “F (2,18) = 2.606, p = .0003”

  • Doesn’t add upF(2, 20) = 1.096reported p = .0017 · recomputed p = .353The two fall on opposite sides of .05

    “F (2,20) = 1.096, p = .0017”

  • Doesn’t add upF(2, 18) = 2.606reported p = .0003 · recomputed p = .101The two fall on opposite sides of .05

    “F (2, 18) = 2.606, p = .0003”

  • Doesn’t add upF(2, 16) = 8.395reported p < .0001 · recomputed p = .003

    “F (2,16) = 8.395, p < .0001”

  • Doesn’t add upF(2, 20) = 10.84reported p = .0012 · recomputed p < .001

    “F (2,20) = 10.84, p = .0012”

  • Doesn’t add upF(2, 16) = 8.395reported p < .0001 · recomputed p = .003

    “F (2, 16) = 8.395, p < .0001”

  • Doesn’t add upF(2, 20) = 10.84reported p = .0012 · recomputed p < .001

    “F (2, 20) = 10.84, p = .0012”

  • ConsistentF(2, 20) = 40.14reported p < .0001 · recomputed p < .001

    “F (2,20) = 40.14, p < .0001”

  • ConsistentF(1.998, 39.96) = 11.9reported p < .0001 · recomputed p < .001

    “F (1.998, 39.96) = 11.90, p < .0001”

  • ConsistentF(4, 40) = 2.227reported p = .0833 · recomputed p = .083

    “F (4,40) = 2.227, p = .0833”

  • ConsistentF(2, 20) = 8.11reported p = .0026 · recomputed p = .003

    “F (2,20) = 8.110, p = .0026”

  • ConsistentF(2, 19) = 15.72reported p < .0001 · recomputed p < .001

    “F (2,19) = 15.72, p < .0001”

  • ConsistentF(2, 20) = 14.63reported p = .0001 · recomputed p < .001

    “F (2,20) = 14.63, p = .0001”

  • ConsistentF(2, 20) = 8.509reported p = .0021 · recomputed p = .002

    “F (2,20) = 8.509, p = .0021”

  • ConsistentF(2, 20) = 16.3reported p < .0001 · recomputed p < .001

    “F (2,20) = 16.30, p < .0001”

  • ConsistentF(2, 14) = 3.473reported p = .0596 · recomputed p = .060

    “F (2,14) = 3.473, p = .0596”

  • ConsistentF(2, 19) = 16.34reported p < .0001 · recomputed p < .001

    “F (2,19) = 16.34, p < .0001”

  • ConsistentF(2, 16) = 34.55reported p < .0001 · recomputed p < .001

    “F (2,16) = 34.55, p < .0001”

  • ConsistentF(2, 15) = 2.099reported p = .1571 · recomputed p = .157

    “F (2,15) = 2.099, p = .1571”

  • ConsistentF(2, 20) = 40.14reported p < .0001 · recomputed p < .001

    “F (2, 20) = 40.14, p < .0001”

  • ConsistentF(4, 40) = 2.227reported p = .0833 · recomputed p = .083

    “F (4, 40) = 2.227, p = .0833”

  • ConsistentF(2, 20) = 8.11reported p = .0026 · recomputed p = .003

    “F (2, 20) = 8.110, p = .0026”

  • ConsistentF(2, 20) = 14.63reported p = .0001 · recomputed p < .001

    “F (2, 20) = 14.63, p = .0001”

  • ConsistentF(2, 20) = 8.509reported p = .0021 · recomputed p = .002

    “F (2, 20) = 8.509, p = .0021”

  • ConsistentF(2, 20) = 16.3reported p < .0001 · recomputed p < .001

    “F (2, 20) = 16.30, p < .0001”

  • ConsistentF(2, 19) = 16.34reported p < .0001 · recomputed p < .001

    “F (2, 19) = 16.34, p < .0001”

  • ConsistentF(2, 16) = 34.55reported p < .0001 · recomputed p < .001

    “F (2, 16) = 34.55, p < .0001”

  • ConsistentF(2, 15) = 2.099reported p = .1571 · recomputed p = .157

    “F (2, 15) = 2.099, p = .1571”

28 of 28

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.

  • A nonsignificant p value is described as significant

    Major

    The paper calls the treatment-group effect on TSPO-positive cell counts significant at p = 0.0596, although its stated significance threshold is p < 0.05.

1 of 1

Cites

Flagged first, then by citations

Research works printed but not linked: 1

  1. Betlazar C, Harrison-Brown M, Middleton RJ, Banati R, Liu GJ. Cellular sources and regional variations in the expression of the neuroinflammatory marker translocator protein (TSPO) in the normal brain. Int J Mol Sci. 2018;19:2707. · DOI · Paper page

Cited by

Sorted by citations

Related

Authors
Vicente-Rodríguez, M., Mancuso, R., Peris-Yague, A., Simmons, C., NIMA Consortium, Wlazly, D., Dickinson, A., Foster, A., Knight, C., Leckey, C., Morgan, P., Morgan, A., O’Hagan, C., Touchard, S., Khan, S., Murphy, P., Parker, C., Patel, J., Richardson, J., Acton, P., Austin, N., Bhattacharya, A., Carruthers, N., de Boer, P., Drevets, W., Isaac, J., Jones, D., Kemp, J., Kolb, H., Nye, J., Wittenberg, G., Barker, G., Bogdanova, A., Byrom, H., Cattaneo, A., Enache, D., Gee, T., Hastings, C., Kose, M., Lombardo, G., Mariani, N., McLaughlin, A., Mondelli, V., Nettis, M., Nikkheslat, N., Pariante, C., Randall, K., Schubert, J., Sforzini, L., Sheridan, H., Singh, N., Van Loo, V., Veronese, M., Wood, T., Worrell, C., Zajkowska, Z., Campbell, B., Egebjerg, J., Eriksson, H., Gastambide, F., Adams, K. H., Jeggo, R., Moeller, T., Nelson, B., Plath, N., Thomsen, C., Pederson, J. T., Zorn, S., Deith, C., Farmer, S., McClean, J., McPherson, A., Penandes, N., Scouller, P., Sutherland, M., Attenburrow, M. J., Benjamin, J., Jones, H., Mada, F., Oladejo, A., Smith, K., Balice-Gordon, R., Binneman, B., Duerr, J., Fullerton, T., Goli, V., Hughes, Z., Piro, J., Samad, T., Sporn, J., Hoskins, L., Kohn, C., Wilcock, L., Aigbirhio, F., Bhatti, J., Bullmore, E., Chamberlain, S., Correia, M., Crofts, A., Fryer, T., Graves, M., Hatton, A., Kitzbichler, M., Lynall, M.-E., Maurice, C., O’Donnell, C., Pointon, L., Hyslop, P. S. G., Turner, L., Vertes, P., Widmer, B., Williams, G., Cavanagh, J., McColl, A., Shaw, R., Boddeke, E., Baird, A., Clare, S., Cowen, P., Huang, I.-S., Hurley, S., Nevado-Holgado, A., Ribe, E., Vyas, A., Winchester, L., Cleal, M., Gomez-Nicola, D., Perry, H., Cercignani, M., Clarke, C., Colasanti, A., Harrison, N., Murray, R., O’Connor, J., Mount, H., Gómez-Nicola, D., Perry, V. H., Turkheimer, F., Lovestone, S., Parker, C. A., & Cash, D.
Affiliations
  • University of Antwerp (BE)
  • King's College London (GB)
  • Universidad San Pablo CEU (ES)
  • Janssen (United Kingdom) (GB)
ORCID
Venue
Journal of Neuroinflammation
Publisher
Springer Science and Business Media LLC
Published
2023-04-09
Volume
20
Issue
1
Pages
92-92
ISSN
1742-2094 (ISSN-L)
DOI
10.1186/s12974-023-02769-y
PMC
PMC10084680
OpenAlex
W4362720149
Semantic Scholar
e5b5069339142405404d1d80e612c8d8296df5e4
Cited by
17
References
63
Citations by year
2026: 3 · 2025: 11 · 2024: 1 · 2023: 2
FWCI
1.40
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Neuroinflammation (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • ePrints Soton (University of Southampton) (acceptedVersion, other-oa)
  • Archivio Istituzionale della Ricerca (Universita Degli Studi Di Milano) (submittedVersion, cc-by)
  • PubMed Central (submittedVersion, cc-by)
  • Apollo (University of Cambridge) (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Figshare (submittedVersion, cc-by)
  • Research Portal (King's College London) (submittedVersion, other-oa)
  • CEU Repositorio Institucional (Fundación Universitaria San Pablo CEU) (submittedVersion, cc-by-nc-nd)
  • Padua Research Archive (University of Padova) (submittedVersion, cc-by)
Funders
GlaxoSmithKline
Topics
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Prion Diseases and Protein Misfolding
  • Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
Keywords
translocator protein (TSPO), neuroinflammation, prion disease, CSF1R inhibitor, microglia, neurons, astrocytes, neurodegeneration, hippocampus, neuroinflammation biomarkers, positron emission tomography
MeSH terms
Neuroinflammatory Diseases, Animals, Brain, Macrophages, Neurons, Biomarkers, Prion Diseases, Microglia, Receptors, GABA, Neurodegenerative Diseases, Endothelial Cells, Positron-Emission Tomography, Mice

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 2026-10-18.

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