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Cited by 312Extracellular vesicles in diseaseNeuroinflammation and Neurodegeneration MechanismsNeuroscience and Neuropharmacology Research

Glia-to-neuron transfer of miRNAs via extracellular vesicles: a new mechanism underlying inflammation-induced synaptic alterations

Open access
Prada, Ilaria, Gabrielli, Martina, Turola, Elena, Iorio, Alessia, D’Arrigo, Giulia, Parolisi, Roberta, De Luca, Mariacristina, Pacifici, Marco +8 more·2018·Acta Neuropathologica
DOI 10.1007/s00401-017-1803-xPMC PMC5978931
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Abstract

Recent evidence indicates synaptic dysfunction as an early mechanism affected in neuroinflammatory diseases, such as multiple sclerosis, which are characterized by chronic microglia activation. However, the mode(s) of action of reactive microglia in causing synaptic defects are not fully understood. In this study, we show that inflammatory microglia produce extracellular vesicles (EVs) which are enriched in a set of miRNAs that regulate the expression of key synaptic proteins. Among them, miR-146a-5p, a microglia-specific miRNA not present in hippocampal neurons, controls the expression of presynaptic synaptotagmin1 (Syt1) and postsynaptic neuroligin1 (Nlg1), an adhesion protein which play a crucial role in dendritic spine formation and synaptic stability. Using a Renilla-based sensor, we provide formal proof that inflammatory EVs transfer their miR-146a-5p cargo to neuron. By western blot and immunofluorescence analysis we show that vesicular miR-146a-5p suppresses Syt1 and Nlg1 expression in receiving neurons. Microglia-to-neuron miR-146a-5p transfer and Syt1 and Nlg1 downregulation do not occur when EV-neuron contact is inhibited by cloaking vesicular phosphatidylserine residues and when neurons are exposed to EVs either depleted of miR-146a-5p, produced by pro-regenerative microglia, or storing inactive miR-146a-5p, produced by cells transfected with an anti-miR-146a-5p. Morphological analysis reveals that prolonged exposure to inflammatory EVs leads to significant decrease in dendritic spine density in hippocampal neurons in vivo and in primary culture, which is rescued in vitro by transfection of a miR-insensitive Nlg1 form. Dendritic spine loss is accompanied by a decrease in the density and strength of excitatory synapses, as indicated by reduced mEPSC frequency and amplitude. These findings link inflammatory microglia and enhanced EV production to loss of excitatory synapses, uncovering a previously unrecognized role for microglia-enriched miRNAs, released in association to EVs, in silencing of key synaptic genes.

AI snapshot

Written from the paper’s full text.

TL;DR

Inflammatory microglia release extracellular vesicles enriched in miRNAs, including miR-146a-5p, which transfer to hippocampal neurons and suppress the synaptic proteins Syt1 and Nlg1. This EV-mediated miRNA transfer reduces dendritic spine and excitatory synapse density and weakens mEPSCs, linking reactive glia to synaptic loss in neuroinflammation.

Key findings

  • EVs from reactive microglia are enriched in miR-146a-5p, miR-181a, and miR-223; miR-146a-5p was upregulated in M1-EVs versus M2-EVs (fold change 6.92) and in Aβ-EVs versus M2-EVs (22.70), and was present in CSF EVs pooled from 10 MS patients but undetectable in cultured hippocampal neurons.
  • EV-to-neuron miRNA transfer was shown with Renilla reporters and cel-miR-39; transfer was blocked by annexin-V cloaking of phosphatidylserine or by donor anti-miR-146a-5p. Inflammatory EVs reduced Syt1 immunostaining by 50% at 48 h and decreased Syt1 and Nlg1 by western blot at 72 h (Syt1 P = 0.008; Nlg1 P = 0.001), without changing MAP2, SNAP25, or PSD95.
  • Inflammatory MVs reduced dendritic spine density and synaptic contacts; a miR-insensitive Nlg1 rescued spine loss. In vivo CA1 infusion of M1-EVs decreased spine density by ~30% compared with intact, vehicle-, and M2-EV-injected mice.
  • mEPSC frequency and amplitude were reduced in MV-treated neurons (frequency P = 0.024; amplitude P = 0.036; control mean frequency 1.537 ± 0.356 Hz; control mean amplitude 25.445 ± 1.503 pA).

Methods

  • Primary rat mixed glial cultures, pure microglia, astrocytes, and hippocampal neurons were used; microglia were polarized with Th1 cytokines, IL-4, or Aβ 1–42, and astrocytes with LPS. EVs were isolated by differential centrifugation and quantified by tunable resistive pulse sensing.
  • The study combined miRNA profiling/qPCR, dual luciferase reporters, western blot, immunofluorescence, dendritic spine/synapse imaging, electrophysiology, in vivo osmotic minipump delivery into mouse CA1 with Golgi–Cox staining, and optical tweezers with calcein/R18 membrane-fusion assays.
  • Data are presented as mean ± SE; statistical significance was assessed with tests specified in figure legends, with P < 0.05 considered significant.

Limitations

  • Stated: the minimum number of inflammatory EVs required to alter neuronal miR-146a-5p levels and suppress target genes remains undefined; additional experiments are needed to clarify whether miR-223 alters GluR2 and whether synaptic dysfunction results from combined miRNA actions. The authors also note possible contributions of non-EV secretory molecules, since SNAP25 was reduced in astrocyte co-cultures and PSD95 tended to decrease with inflammatory microglia.
  • Evident: in vivo spine analysis used n = 3 mice per condition; CSF EV analysis pooled samples from 10 MS patients without a non-MS comparison group; some assays used very small n (e.g., RNase protection n = 2); and the in vivo experiments infused rat microglial EVs into mouse hippocampus.

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

Integrity signals

Registry

Crossref · 10.1007/s00401-017-1803-x

Cites

88 of 88 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

1 of 71 cited sources checked · 0 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    Microglia-derived MVs affect the presynaptic compartment causing an excitation-inhibition unbalance.

    “Our previous studies have shown that microglia-derived MVs affect the presynaptic compartment causing an excitation–inhibition unbalance.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [62] Riganti L, Antonucci F, Gabrielli M, Prada I, Giussani P, Viani P, Valtorta F, Menna E, Matteoli M, Verderio C (2016) Sphingosine-1-Phosphate (S1P) impacts presynaptic functions by regulating synapsin I localization in the presynaptic compartment. J Neurosci 36:4624–4634. https://doi.org/10.1523/JNEUROSCI.3588-15.2016 source
    • cites [1] Antonucci F, Turola E, Riganti L, Caleo M, Gabrielli M, Perrotta C, Novellino L, Clementi E, Giussani P, Viani P et al (2012) Microvesicles released from microglia stimulate synaptic activity via enhanced sphingolipid metabolism. EMBO J 31:1231–1240. https://doi.org/10.1038/emboj.2011.489 source

      The Discussion reports that microglia-derived MVs mainly act at presynaptic excitatory synapses, increasing vesicle release probability. It says the observed change in visual-cortex receptive fields can typically be explained by altered excitation–inhibition balance, but does not establish that MVs caused such an imbalance. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
  • Findingown data

    Syt1 and Nlg1 downregulation does not occur when neurons are exposed to EVs depleted of miR-146a-5p.

    “and when neurons are exposed to EVs either depleted of miR-146a-5p, produced by pro-regenerative microglia, or storing inactive miR-146a-5p, produced by cells transfected with an anti-miR-146a-5p.”
    Quote in the paperSubstantial claim
  • Findingown data

    Spontaneous miniature synaptic currents are diminished in receiving neurons.

    “Notably, protein levels of Syt1 and Nlg1 are decreased, the density of dendritic spines and excitatory synapses is reduced, and spontaneous miniature synaptic currents are diminished in receiving neurons, indicating important synaptic alterations.”
    Quote in the paperSubstantial claim
  • Findingown data

    miR-146a-5p represses translation of presynaptic Syt1 and postsynaptic Nlg1.

    “We show that miR-146a-5p, which represses translation of presynaptic Syt1 and postsynaptic Nlg1, is upregulated in EVs secreted from reactive microglia either exposed to inflammatory or degenerative stimuli, and is delivered to neurons upon EV fusion with the plasma membrane.”
    Quote in the paperSubstantial claim
  • Findingown data

    miR-146a-5p was undetectable in cultured hippocampal neurons unlike miR-181a and miR-223.

    “Conversely, miR-146a-5p was undetectable in cultured hippocampal neurons unlike miR-181a and miR-223 (Fig. 2d).”
    Quote in the paperSubstantial claim
  • May combine two claims. 50
  • Classified as probably not a substantial claim. 14

  • May combine two claims.

    Findingown data

    Decreased Syt1 immunoreactivity and decreased spine density occur in neurons transfected with miR-146a-5p.

    “(3) decreased Syt1 immunoreactivity and decreased spine density in neurons transfected with miR-146a-5p.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    EV-mediated shuttling of miRNAs between reactive glia and neurons and consequent downregulation of their synaptic targets occurs.

    “We have provided formal proof for EV-mediated shuttling of miRNAs between reactive glia and neurons and consequent downregulation of their synaptic targets in a number of ways:”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Q-PCR analysis confirmed that miR-146a-5p is present in cultured astrocytes and MVs thereof, more abundantly upon activation with LPS.

    “Q-PCR analysis confirmed that miR-146a-5p is present in cultured astrocytes and MVs thereof, more abundantly upon activation with LPS (Fig. 2e)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Prolonged exposure to inflammatory MVs did not alter the expression of genes not predicted to be targeted by miR-146a-5p (SNAP25, PSD95, and MAP2), indicating the absence of synaptic or neuronal damage.

    “Prolonged exposure to inflammatory MVs did not alter the expression of genes not predicted to be targeted by miR-146a-5p (SNAP25, PSD95, and MAP2), indicating the absence of synaptic or neuronal damage (Fig. 3c, d).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    MVs and exosomes produced by non-stimulated microglia contained 78 and 71 miRNAs, respectively, with 21 unique in MVs, 14 unique in exosomes, and 57 common.

    “We found a total of 78 and 71 miRNAs, respectively, in MVs and exosomes produced by non-stimulated (NS) microglia, of which 21 were unique in MVs and 14 unique in exosomes (Fig. 1b), with a strong overlap between the two EV populations (57 common miRNAs).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Inflammatory MVs caused a significant decrease in mEPSC frequency and amplitude, indicating a decrease in synaptic strength.

    “Inflammatory MVs caused a significant decrease in mEPSC frequency (Fig. 5b) and amplitude (Fig. 5c), indicating a decrease in synaptic strength.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Reduced Syt1 and Nlg1 expression occurs in neurons incubated with miR146a-enriched MVs (derived from inflammatory microglia or astrocytes), but not miR-146a-5p-depleted MVs (released from pro-regenerative microglia) or MVs storing inactive miR-146a-5p (produced by astrocytes treated with a miR-146a-5p inhibitor).

    “(1) reduced Syt1 and Nlg1 expression in neurons incubated with miR146a-enriched MVs (derived from inflammatory microglia or astrocytes), but not miR-146a-5p-depleted MVs (released from pro-regenerative microglia) or MVs storing inactive miR-146a-5p (produced by astrocytes treated with a miR-146a-5p inhibitor);”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Unstimulated microglia (1 × 10^6 cells) secrete approximately 1 × 10^8 exosomes per hour, with mean diameter 92 ± 5.28 nm.

    “and ~ 1 × 108 exosomes (mean diameter = 92 ± 5.28 nm) per hour.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    A large fraction of astrocyte-derived MVs adhered to the cell body of neurons (~74% n = 47) and more than half of microglial MVs was able to bind to neuronal surface (~53% n = 30).

    “A large fraction of astrocyte-derived MVs adhered to the cell body of neurons (~ 74% n = 47) and more than half of microglial MVs was able to bind to neuronal surface (~ 53% n = 30).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    None of the MVs attached to neurons disappeared from the observation field, excluding rapid MV uptake or full fusion with the neuron plasma membrane.

    “Importantly, none of the MVs attached to neurons disappeared from the observation field, thus excluding rapid MV uptake or full fusion with the neuron plasma membrane (PM).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    After contact with neuron cell bodies, MVs showed little displacements around the place of delivery and remained attached to the neuron surface for up to 1-2 h of recording.

    “After contact with neuron cell bodies, MVs showed little displacements around the place of delivery and remained attached to the neuron surface for up to 1–2 h of recording (Fig. 7b).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    MVs secreted from M2 microglia, depleted in miR-146a-5p, did not cause spine loss and rather tended to increase the density of mature (mushroom) spines.

    “Conversely, MVs secreted from M2 microglia, depleted in miR-146a-5p, did not cause spine loss and rather tended to increase the density of mature (mushroom) spines (Fig. 4f).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    48 h after MV addition, presynaptic terminals still displayed intense Syt1 fluorescence and no change of total Syt1 protein levels was detected by western blot analysis.

    “Accordingly, 48 h after MV addition, presynaptic terminals still displayed intense Syt1 fluorescence (Fig. 3a, arrowhead) and no change of total Syt1 protein levels was detected by western blot analysis (not shown).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Density of dendritic spines and excitatory synapses is reduced in receiving neurons.

    “Notably, protein levels of Syt1 and Nlg1 are decreased, the density of dendritic spines and excitatory synapses is reduced, and spontaneous miniature synaptic currents are diminished in receiving neurons, indicating important synaptic alterations.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Unstimulated microglia (1 × 10^6 cells) secrete approximately 3.5 × 10^7 MVs per hour, with mean diameter 182.82 ± 3.35 nm.

    “Quantification by tunable resistive pulse sensing (TRPS) revealed that unstimulated microglia (1 × 106 cells) showed secretion rate of ~ 3.5 × 107 MVs (mean diameter = 182.82 ± 3.35 nm)”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Immunocytochemical staining for Bassoon and Shank2 showed a significant decrease in the percentage of juxtaposed pre- and postsynaptic terminals and in synaptic density in neurons receiving inflammatory MVs.

    “Immunocytochemical staining for the presynaptic active zone marker Bassoon and the postsynaptic density marker Shank2 showed a significant decrease in the percentage of juxtaposed pre- and postsynaptic terminals (Fig. 4d) and in synaptic density (Fig. 4e) in neurons receiving inflammatory MVs.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Glia-to-neuron transfer of miR-146a-5p suppresses translation of a key dendritic protein, Nlg1, and decreases synaptic density and strength.

    “Our study extends and complements these findings by showing that glia-to-neuron transfer of miR-146a-5p suppresses translation of a key dendritic protein, Nlg1, and decreases synaptic density and strength.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    72 h incubation with MVs derived from inflammatory astrocytes caused a significant decrease in spine density, mimicking the reduction induced by transfection with miR-146a-5p-GFP.

    “72 h incubation with MVs derived from inflammatory astrocytes caused a significant decrease in spine density (Fig. 4a), mimicking the reduction induced by transfection with miR-146a-5p-GFP (Fig. 4b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Astrocyte-derived MVs reduced by 50% Syt1 immunostaining 48 h after MV administration to neurons, similarly to direct miR-146a-5p transfection, and the MV effect was rescued by transfection of donor glia with anti-miR-146a-5p.

    “Astrocyte-derived MVs reduced by 50% Syt1 immunostaining 48 h after MV administration to neurons, similarly to direct miR-146a-5p transfection, and the MV effect was rescued by transfection of donor glia with anti-miR-146a-5p (Fig. 3a, b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Reduced spine and synapse density occurs in neurons exposed to miR146a-enriched MVs, derived from either reactive microglia or astrocytes, not occurring when neurons were exposed to miR146a-depleted MVs, derived from IL4-treated microglia.

    “(2) reduced spine and synapse density in neurons exposed to miR146a-enriched MVs, derived from either reactive microglia or astrocytes, not occurring when neurons were exposed to miR146a-depleted MVs, derived from IL4-treated microglia;”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Compared to parental microglia, EVs contained a selection of miRNAs and few vesicle-specific miRNAs not detectable in donor cells.

    “Compared to parental microglia, EVs contained a selection of miRNAs and few vesicle-specific miRNAs, not detectable in donor cells.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Analysis of cell viability revealed similar percentage of calcein-positive and propidium iodide negative viable neurons in control and MVs-treated cultures (% of calcein+/PI− neurons/overall nuclei: ctr = 69.32 ± 1.87; MVs = 81.83 ± 1.95; number of neurons: ctr = 355, MVs = 326; n = 3) further excluding neurotoxic effects.

    “Analysis of cell viability revealed similar percentage of calcein-positive and propidium iodide negative viable neurons in control and MVs-treated cultures (% of calcein+/PI− neurons/overall nuclei: ctr = 69.32 ± 1.87; MVs = 81.83 ± 1.95; number of neurons: ctr = 355, MVs = 326; n = 3) further excluding neurotoxic effects.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    MVs secreted from M2 microglia had no effect on the fraction of juxtaposed pre- and postsynaptic terminals and the density of synaptic contacts.

    “In addition, they had no effect on the fraction of juxtaposed pre- and postsynaptic terminals (Fig. 4g) and the density of synaptic contacts (Fig. 4h).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    miR-146a-5p levels increase in neurons transfected with a specific Renilla reporter in response to inflammatory EVs but not EVs released from anti-miR-146a-5p-treated cells.

    “(2) monitoring miR-146a-5p, a glia-specific miRNA upregulated in EVs produced by inflammatory glia, showed that miR-146a-5p levels increase in neurons transfected with a specific Renilla reporter in response to inflammatory EVs but not EVs released from anti-miR-146a-5p-treated cells;”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Q-PCR analysis showed that some miRNAs were several folds more represented in MVs and exosomes than in donor cells, while others were low abundant in MVs and exosomes than in cells.

    “Q-PCR analysis showed that some miRNAs were several folds more represented in MVs and exosomes than in donor cells, while others were low abundant in MVs and exosomes than in cells (Fig. 1c, Fig. S2).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Dendritic spine destabilization in response to miR-146-enriched MVs was confirmed in neurons treated with MVs released from inflammatory microglia (M1-MVs), where loss of immature (thin) spines mostly accounted for reduced spine density.

    “Dendritic spine destabilization in response to miR-146-enriched MVs was confirmed in neurons treated with MVs released from inflammatory microglia (M1-MVs), where loss of immature (thin) spines mostly accounted for reduced spine density (Fig. 4f).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Overnight exposure to reactive EVs (r-MVs or r-exosomes), but not MVs derived from unstimulated astrocytes, caused a significant reduction in Renilla/firefly luciferase activity in EV-treated neurons compared to control untreated neurons, indicating an increase in neuronal miR-146a-5p levels.

    “Overnight exposure to reactive EVs (r-MVs or r-exosomes), but not MVs derived from unstimulated astrocytes (MVs), caused a significant reduction in Renilla/firefly luciferase activity in EV-treated neurons compared to control untreated neurons, indicating an increase in neuronal miR-146a-5p levels (Fig. 2g).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Transfer of miR-146a-5p via EVs is sufficient to decrease Syt1 and Nlg1 expression, to reduce dendritic spine in vitro and in the CA1 region of the hippocampus, and to decrease synapse density and strength in cultured neurons.

    “We have shown that transfer of miR-146a-5p via EVs is sufficient to decrease Syt1 and Nlg1 expression, to reduce dendritic spine in vitro and in the CA1 region of the hippocampus and to decrease synapse density and strength in cultured neurons, revealing that this miRNA hampers proper synapse stability and function.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Differential expression analysis of M1-EVs versus M2-EVs revealed 21 miRNAs, among which 11 were upregulated and 10 downregulated in M1-EVs.

    “Differential expression analysis of M1-EVs versus M2-EVs revealed 21 miRNAs, among which 11 were upregulated and 10 downregulated in M1-EVs (Fig. 1e; Table 1).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    MV fusion was proven by dequenching of R18 fluorescence at contact sites while calcein retention in MVs suggested the opening of small pores, not permeable to the cytosolic dye.

    “MV fusion was proven by dequenching of R18 fluorescence at contact sites while calcein retention in MVs suggested the opening of small pores, not permeable to the cytosolic dye.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingcitation

    EV fusion also occurs along neuronal processes, where synaptic activity is influenced by EVs.

    “Importantly, EV fusion also occurs along neuronal processes, where synaptic activity is influenced by EVs [1, 62].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [1] Antonucci F, Turola E, Riganti L, Caleo M, Gabrielli M, Perrotta C, Novellino L, Clementi E, Giussani P, Viani P et al (2012) Microvesicles released from microglia stimulate synaptic activity via enhanced sphingolipid metabolism. EMBO J 31:1231–1240. https://doi.org/10.1038/emboj.2011.489 source

      Not checked in the free run.

    • cites [62] Riganti L, Antonucci F, Gabrielli M, Prada I, Giussani P, Viani P, Valtorta F, Menna E, Matteoli M, Verderio C (2016) Sphingosine-1-Phosphate (S1P) impacts presynaptic functions by regulating synapsin I localization in the presynaptic compartment. J Neurosci 36:4624–4634. https://doi.org/10.1523/JNEUROSCI.3588-15.2016 source

  • May combine two claims.

    Findingown data

    A relevant fraction of MVs derived from microglia or astrocytes adhere to neurons, and can therefore be potentially functional as communication vehicles.

    “Using optical tweezers, a unique tool to finely position single EVs on target cells and to study EV-cell interaction individually [59], here we show that a relevant fraction of MVs derived from microglia or astrocytes adhere to neurons, and can therefore be potentially functional as communication vehicles.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingcitation

    Heparan sulfate proteoglycans on the surface of receiving neurons mediate the interaction with only a fraction of MVs, indicating that multiple molecules are likely to participate in MV-neuron contact.

    “Similarly, heparan sulfate proteoglycans [14] on the surface of receiving neurons mediate the interaction with only a fraction of MVs, indicating that multiple molecules are likely to participate in MV–neuron contact.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    gluN2B is a receptor essential to form LTP.

    “Among them, miR-223, which targets the AMPA-selective glutamate receptor 2 (GluR2) and the NMDA glutamate receptor gluN2B [33] and miR-181a which regulates only gluN2B [66], a receptor essential to form LTP [8, 71].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [8] Bliss TV, Collingridge GL (2013) Expression of NMDA receptor-dependent LTP in the hippocampus: bridging the divide. Mol Brain 6:5. https://doi.org/10.1186/1756-6606-6-5 source

      Not checked in the free run.

    • cites [71] Shipton OA, Paulsen O (2014) GluN2A and GluN2B subunit-containing NMDA receptors in hippocampal plasticity. Philos Trans R Soc Lond B Biol Sci 369:20130163. https://doi.org/10.1098/rstb.2013.0163 source

  • May combine two claims.

    Premisecitation

    miR-146a-5p is a microglia-enriched miRNA, detectable at lesser extent in astrocytes, and it prevents the expression of neuronal genes in neuron progenitors.

    “Among the three miRNAs upregulated in EVs produced by inflammatory microglia, miR-146a-5p is a microglia-enriched miRNA, detectable at lesser extend in astrocytes, and it prevents the expression of neuronal genes in neuron progenitors [39].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [39] Jovicic A, Roshan R, Moisoi N, Pradervand S, Moser R, Pillai B, Luthi-Carter R (2013) Comprehensive expression analyses of neural cell-type-specific miRNAs identify new determinants of the specification and maintenance of neuronal phenotypes. J Neurosci Off J Soc Neurosci 33:5127–5137. https://doi.org/10.1523/JNEUROSCI.0600-12.2013 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    miRNAs are additional active components of inflammatory MVs responsible for delayed destabilization of excitatory synapses and decreased transmission, selectively during chronic inflammation.

    “In this study, we have identified miRNAs as additional active components of inflammatory MVs that are responsible for a delayed destabilization of excitatory synapses and decreased transmission, which selectively occur during chronic inflammation.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The inflammatory stimuli used in the study model chronic brain inflammation relevant to human neuroinflammatory disorders.

  • May combine two claims.

    Premisecitation

    All synaptic targets of miR-146a-5p, miR-181a and miR-223 are conserved between mouse and rat, and Syt1 is a conserved target also in human.

    “All the synaptic targets of miR-146a-5p, miR-181a and miR-223 are conserved between mouse and rat and Syt1 is a conserved target also in human, as documented by Saba and coworkers [66] and the miRNA databases TargetScan v 7.1 and microRNA.org.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Saba and coworkers [66]

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

    • cites TargetScan v 7.1

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

    • cites microRNA.org

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

  • May combine two claims.

    Interpretationown data

    miR-146a-5p-enriched EVs released from inflammatory microglia promote dendritic spine loss in vivo, likely through downregulation of Nlg1, which is a validated miR-146a-5p target in both rat and mouse brain.

    “This result confirms in vivo that miR-146a-5p-enriched EVs released from inflammatory microglia promote dendritic spine loss, likely through downregulation of Nlg1, which is a validated miR-146a-5p target in both rat and mouse brain.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    miR-223 targets the AMPA-selective glutamate receptor 2 (GluR2) and the NMDA glutamate receptor gluN2B.

    “Among them, miR-223, which targets the AMPA-selective glutamate receptor 2 (GluR2) and the NMDA glutamate receptor gluN2B [33] and miR-181a which regulates only gluN2B [66], a receptor essential to form LTP [8, 71].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [33] Harraz MM, Eacker SM, Wang X, Dawson TM, Dawson VL (2012) MicroRNA-223 is neuroprotective by targeting glutamate receptors. Proc Natl Acad Sci USA 109:18962–18967. https://doi.org/10.1073/pnas.1121288109 source

  • May combine two claims.

    Interpretationown data

    MVs secreted from inflammatory glial cells deliver bioactive miR-146a-5p to neurons and modulate the expression of its synaptic targets according to their miR-146a-5p cargo.

    “Taken together, these data indicate that MVs secreted from inflammatory glial cells deliver bioactive miR-146a-5p to neurons and modulate the expression of its synaptic targets according to their miR-146a-5p cargo.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Previous evidence indicates that miR-146a-5p, miR-181a and miR-223 are dysregulated in patients affected by neuroinflammatory and neurodegenerative diseases.

    “Previous evidence indicates that miR-146a-5p, miR-181a and miR-223 are dysregulated in patients affected by neuroinflammatory [36] and neurodegenerative diseases [22, 37, 60], that are characterized by high production of myeloid EVs [38, 78].”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [36] Jagot F, Davoust N (2016) Is it worth considering circulating microRNAs in multiple sclerosis? Front Immunol 7:129. https://doi.org/10.3389/fimmu.2016.00129 source
    • cites [22] Ding H, Huang Z, Chen M, Wang C, Chen X, Chen J, Zhang J (2016) Identification of a panel of five serum miRNAs as a biomarker for Parkinson’s disease. Parkinsonism Relat Disord 22:68–73. https://doi.org/10.1016/j.parkreldis.2015.11.014 source
    • cites [37] Jia LH, Liu YN (2016) Downregulated serum miR-223 servers as biomarker in Alzheimer’s disease. Cell Biochem Funct 34:233–237. https://doi.org/10.1002/cbf.3184 source
    • cites [60] Properzi F, Ferroni E, Poleggi A, Vinci R (2015) The regulation of exosome function in the CNS: implications for neurodegeneration. Swiss Med Wkly 145:w14204. https://doi.org/10.4414/smw.2015.14204 source

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

    Premisecitation

    Consolidated evidence describes miR-146a-5p upregulation both at MS lesions and in CSF from patients with Alzheimer's diseases.

    “In particular, consolidated evidence describes miR-146a-5p upregulation both at MS lesions [40] and in CSF from patients with Alzheimer’s diseases [32, 61].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [40] Junker A, Krumbholz M, Eisele S, Mohan H, Augstein F, Bittner R, Lassmann H, Wekerle H, Hohlfeld R, Meinl E (2009) MicroRNA profiling of multiple sclerosis lesions identifies modulators of the regulatory protein CD47. Brain J Neurol 132:3342–3352. https://doi.org/10.1093/brain/awp300 source
    • cites [32] Grasso M, Piscopo P, Confaloni A, Denti MA (2014) Circulating miRNAs as biomarkers for neurodegenerative disorders. Molecules 19:6891–6910. https://doi.org/10.3390/molecules19056891 source
    • cites [61] Reddy PH, Tonk S, Kumar S, Vijayan M, Kandimalla R, Kuruva CS, Reddy AP (2017) A critical evaluation of neuroprotective and neurodegenerative MicroRNAs in Alzheimer’s disease. Biochem Biophys Res Commun 483:1156–1165. https://doi.org/10.1016/j.bbrc.2016.08.067 source

  • May combine two claims.

    Premisecitation

    miR-146a-5p targets the postsynaptic cell-adhesion molecule neuroligin1 (Nlg1), the dendritic protein MAP1A and the synaptic vesicle protein synaptotagmin1 (Syt1).

    “MiR-146a-5p targets the postsynaptic cell-adhesion molecule neuroligin1 (Nlg1), the dendritic protein MAP1A and the synaptic vesicle protein synaptotagmin1 (Syt1) [39];”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [39] Jovicic A, Roshan R, Moisoi N, Pradervand S, Moser R, Pillai B, Luthi-Carter R (2013) Comprehensive expression analyses of neural cell-type-specific miRNAs identify new determinants of the specification and maintenance of neuronal phenotypes. J Neurosci Off J Soc Neurosci 33:5127–5137. https://doi.org/10.1523/JNEUROSCI.0600-12.2013 source

      Not checked in the free run.

  • May combine two claims.

    Study design and conductcitation

    EVs isolated under short ATP stimulation are negative for apoptotic markers and are not contaminated by intracellular organelles derived from damaged cells.

    “Our previous evidence indicates that EVs isolated under short ATP stimulation are negative for apoptotic markers and are not contaminated by intracellular organelles derived from damaged cells, as indicated by electron microscopy [6, 78] and western blot analysis [29].”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [6] Bianco F, Perrotta C, Novellino L, Francolini M, Riganti L, Menna E, Saglietti L, Schuchman EH, Furlan R, Clementi E et al (2009) Acid sphingomyelinase activity triggers microparticle release from glial cells. EMBO J 28:1043–1054. https://doi.org/10.1038/emboj.2009.45 source
    • cites [78] Verderio C, Muzio L, Turola E, Bergami A, Novellino L, Ruffini F, Riganti L, Corradini I, Francolini M, Garzetti L et al (2012) Myeloid microvesicles are a marker and therapeutic target for neuroinflammation. Ann Neurol 72:610–624. https://doi.org/10.1002/ana.23627 source
    • cites [29] Gabrielli M, Battista N, Riganti L, Prada I, Antonucci F, Cantone L, Matteoli M, Maccarrone M, Verderio C (2015) Active endocannabinoids are secreted on extracellular membrane vesicles. EMBO Rep 16:213–220. https://doi.org/10.15252/embr.201439668 source

  • May combine two claims.

    Premisecitation

    MVs acutely enhance glutamate release while suppress spontaneous GABA transmission in both hippocampal cultures and in the visual cortex.

    “MVs acutely enhance glutamate release while suppress spontaneous GABA transmission in both hippocampal cultures [62] and in the visual cortex [1] and their action is due to the lipid components of MVs.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [62] Riganti L, Antonucci F, Gabrielli M, Prada I, Giussani P, Viani P, Valtorta F, Menna E, Matteoli M, Verderio C (2016) Sphingosine-1-Phosphate (S1P) impacts presynaptic functions by regulating synapsin I localization in the presynaptic compartment. J Neurosci 36:4624–4634. https://doi.org/10.1523/JNEUROSCI.3588-15.2016 source
    • cites [1] Antonucci F, Turola E, Riganti L, Caleo M, Gabrielli M, Perrotta C, Novellino L, Clementi E, Giussani P, Viani P et al (2012) Microvesicles released from microglia stimulate synaptic activity via enhanced sphingolipid metabolism. EMBO J 31:1231–1240. https://doi.org/10.1038/emboj.2011.489 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    These findings link inflammatory microglia and enhanced EV production to loss of excitatory synapses.

    “These findings link inflammatory microglia and enhanced EV production to loss of excitatory synapses”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Nlg1 is a postsynaptic protein selectively expressed at excitatory synapses which is known to regulate spine formation and synapse stability through binding to presynaptic neurexins.

    “Nlg1 is a postsynaptic protein selectively expressed at excitatory synapses which is known to regulate spine formation and synapse stability through binding to presynaptic neurexins [16, 70].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [16] Colgan LA, Yasuda R (2014) Plasticity of dendritic spines: subcompartmentalization of signaling. Annu Rev Physiol 76:365–385. https://doi.org/10.1146/annurev-physiol-021113-170400 source

      Not checked in the free run.

    • cites [70] Shipman SL, Nicoll RA (2012) Dimerization of postsynaptic neuroligin drives synaptic assembly via transsynaptic clustering of neurexin. Proc Natl Acad Sci USA 109:19432–19437. https://doi.org/10.1073/pnas.1217633109 source

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

    Premisecitation

    miR-146a-5p, miR-181a and miR-223 have been previously shown to be dysregulated in the CNS, plasma, and/or immune cells of MS patients.

    “Consistently, miR-146a-5p, miR-181a and miR-223 have been previously shown to be dysregulated in the CNS, plasma, and/or immune cells of MS patients [36] but also implicated in neurodegenerative diseases [22, 32, 37, 61].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [36] Jagot F, Davoust N (2016) Is it worth considering circulating microRNAs in multiple sclerosis? Front Immunol 7:129. https://doi.org/10.3389/fimmu.2016.00129 source

  • Classified as probably not a substantial claim.

    Premisecitation

    Inflammatory cytokines TNF-α and IL-1β play a critical role in chronic neuroinflammatory disorders by modulating the expression and/or properties of synaptic channels.

    “inflammatory cytokines, i.e. TNF-α and IL-1β, play a critical role in chronic neuroinflammatory disorders, by modulating the expression and/or properties of synaptic channels”
    Quote in the paperSubstantial claim0 of 5 sources checked
    • cites [12] Centonze D, Muzio L, Rossi S, Cavasinni F, De Chiara V, Bergami A, Musella A, D’Amelio M, Cavallucci V, Martorana A et al (2009) Inflammation triggers synaptic alteration and degeneration in experimental autoimmune encephalomyelitis. J Neurosci Off J Soc Neurosci 29:3442–3452. https://doi.org/10.1523/JNEUROSCI.5804-08.2009 source
    • cites [55] Pascual O, Achour SB, Rostaing P, Triller A, Bessis A (2012) Microglia activation triggers astrocyte-mediated modulation of excitatory neurotransmission. Proc Natl Acad Sci USA 109:E197–E205. https://doi.org/10.1073/pnas.1111098109 source

      Not checked in the free run.

    • cites [65] Rossi S, Muzio L, De Chiara V, Grasselli G, Musella A, Musumeci G, Mandolesi G, De Ceglia R, Maida S, Biffi E et al (2011) Impaired striatal GABA transmission in experimental autoimmune encephalomyelitis. Brain Behav Immun 25:947–956. https://doi.org/10.1016/j.bbi.2010.10.004 source
    • cites [79] Viviani B, Bartesaghi S, Gardoni F, Vezzani A, Behrens MM, Bartfai T, Binaglia M, Corsini E, Di Luca M, Galli CL et al (2003) Interleukin-1beta enhances NMDA receptor-mediated intracellular calcium increase through activation of the Src family of kinases. J Neurosci Off J Soc Neurosci 23:8692–8700 source
    • cites [81] Weber A, Wasiliew P, Kracht M (2010) Interleukin-1 (IL-1) pathway. Sci Signal 3:cm1. https://doi.org/10.1126/scisignal.3105cm1 source

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

    Premisecitation

    Neuron depolarization induces release of EVs containing high amounts of miRNAs.

    “Neuron depolarization has been shown to induce release of EVs containing high amounts of miRNAs, and miRNA elimination has been proposed to promote rapid translation of proteins required for synaptic plasticity [31].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] Goldie BJ, Dun MD, Lin M, Smith ND, Verrills NM, Dayas CV, Cairns MJ (2014) Activity-associated miRNA are packaged in Map1b-enriched exosomes released from depolarized neurons. Nucleic Acids Res 42:9195–9208. https://doi.org/10.1093/nar/gku594 source

  • May combine two claims.

    Premisecitation

    Multiple sclerosis (MS) is a synaptic disorder characterized by chronic microglia activation and enhanced EV secretion with early onset compared to other neurodegenerative diseases.

    “Multiple sclerosis (MS) is a synaptic disorder characterized by chronic microglia activation and enhanced EV secretion [78] with early onset compared to other neurodegenerative diseases.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [78] Verderio C, Muzio L, Turola E, Bergami A, Novellino L, Ruffini F, Riganti L, Corradini I, Francolini M, Garzetti L et al (2012) Myeloid microvesicles are a marker and therapeutic target for neuroinflammation. Ann Neurol 72:610–624. https://doi.org/10.1002/ana.23627 source

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

    Premisecitation

    miRNA elimination promotes rapid translation of proteins required for synaptic plasticity.

    “Neuron depolarization has been shown to induce release of EVs containing high amounts of miRNAs, and miRNA elimination has been proposed to promote rapid translation of proteins required for synaptic plasticity [31].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] Goldie BJ, Dun MD, Lin M, Smith ND, Verrills NM, Dayas CV, Cairns MJ (2014) Activity-associated miRNA are packaged in Map1b-enriched exosomes released from depolarized neurons. Nucleic Acids Res 42:9195–9208. https://doi.org/10.1093/nar/gku594 source

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

    Premisecitation

    miR-146a-5p, miR-181a and miR-223 have been implicated in neurodegenerative diseases.

    “Consistently, miR-146a-5p, miR-181a and miR-223 have been previously shown to be dysregulated in the CNS, plasma, and/or immune cells of MS patients [36] but also implicated in neurodegenerative diseases [22, 32, 37, 61].”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [22] Ding H, Huang Z, Chen M, Wang C, Chen X, Chen J, Zhang J (2016) Identification of a panel of five serum miRNAs as a biomarker for Parkinson’s disease. Parkinsonism Relat Disord 22:68–73. https://doi.org/10.1016/j.parkreldis.2015.11.014 source
    • cites [32] Grasso M, Piscopo P, Confaloni A, Denti MA (2014) Circulating miRNAs as biomarkers for neurodegenerative disorders. Molecules 19:6891–6910. https://doi.org/10.3390/molecules19056891 source
    • cites [37] Jia LH, Liu YN (2016) Downregulated serum miR-223 servers as biomarker in Alzheimer’s disease. Cell Biochem Funct 34:233–237. https://doi.org/10.1002/cbf.3184 source
    • cites [61] Reddy PH, Tonk S, Kumar S, Vijayan M, Kandimalla R, Kuruva CS, Reddy AP (2017) A critical evaluation of neuroprotective and neurodegenerative MicroRNAs in Alzheimer’s disease. Biochem Biophys Res Commun 483:1156–1165. https://doi.org/10.1016/j.bbrc.2016.08.067 source

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

    Premisecitation

    Silencing activity of miR-146a-5p decreases hyperexcitability-underlying seizures in a model of epileptogenesis.

    “Its silencing activity has recently been shown to decrease hyperexcitability-underlying seizures in a model of epileptogenesis, where gliosis and inflammatory cytokines are major pathogenic factors [34].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [34] Iori V, Iyer AM, Ravizza T, Beltrame L, Paracchini L, Marchini S, Cerovic M, Hill C, Ferrari M, Zucchetti M et al (2017) Blockade of the IL-1R1/TLR4 pathway mediates disease-modification therapeutic effects in a model of acquired epilepsy. Neurobiol Dis 99:12–23. https://doi.org/10.1016/j.nbd.2016.12.007 source

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

    Premisecitation

    miR-181a regulates only gluN2B.

    “Among them, miR-223, which targets the AMPA-selective glutamate receptor 2 (GluR2) and the NMDA glutamate receptor gluN2B [33] and miR-181a which regulates only gluN2B [66], a receptor essential to form LTP [8, 71].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [66] Saba R, Storchel PH, Aksoy-Aksel A, Kepura F, Lippi G, Plant TD, Schratt GM (2012) Dopamine-regulated microRNA MiR-181a controls GluA2 surface expression in hippocampal neurons. Mol Cell Biol 32:619–632. https://doi.org/10.1128/MCB.05896-11 source

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

    Premisecitation

    Inflammatory cytokines TNF-α and IL-1β regulate the expression of molecules critical for synaptic plasticity, including CREB or cofilin.

    “and regulating the expression of molecules critical for synaptic plasticity, including CREB or cofilin”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [73] Tong L, Prieto GA, Kramar EA, Smith ED, Cribbs DH, Lynch G, Cotman CW (2012) Brain-derived neurotrophic factor-dependent synaptic plasticity is suppressed by interleukin-1beta via p38 mitogen-activated protein kinase. J Neurosci Off J Soc Neurosci 32:17714–17724. https://doi.org/10.1523/JNEUROSCI.1253-12.2012 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Sustained exposure of neurons to miR-146a-5p-enriched MVs causes over-inhibition of synaptic function and excessive synapse destabilization, leading to pathological loss of synapses, not linked to neuron damage.

    “However, in this study we show that sustained exposure of neurons to miR-146a-5p-enriched MVs causes over-inhibition of synaptic function and excessive synapse destabilization, leading to pathological loss of synapses, not linked to neuron damage, which may underlie synaptic dysfunction occurring in chronic inflammation.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The inflammatory stimuli used in the study model chronic brain inflammation relevant to human neuroinflammatory disorders.

  • May combine two claims.

    Premisecitation

    miR-146a-5p has immunosuppressive and anti-inflammatory functions.

    “Consistent with the idea of a rescue mechanism, miR-146a-5p has well known immunosuppressive and anti-inflammatory functions [20, 35, 76].”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [20] de Candia P, De Rosa V, Casiraghi M, Matarese G (2016) Extracellular RNAs: a secret arm of immune system regulation. J Biol Chem 291:7221–7228. https://doi.org/10.1074/jbc.R115.708842 source
    • cites [35] Iyer A, Zurolo E, Prabowo A, Fluiter K, Spliet WG, van Rijen PC, Gorter JA, Aronica E (2012) MicroRNA-146a: a key regulator of astrocyte-mediated inflammatory response. PLoS One 7:e44789. https://doi.org/10.1371/journal.pone.0044789 source

      Checked before — its earlier verdict is held back until a new check runs.

    • cites [76] van Scheppingen J, Iyer AM, Prabowo AS, Muhlebner A, Anink JJ, Scholl T, Feucht M, Jansen FE, Spliet WG, Krsek P et al (2016) Expression of microRNAs miR21, miR146a, and miR155 in tuberous sclerosis complex cortical tubers and their regulation in human astrocytes and SEGA-derived cell cultures. Glia 64:1066–1082. https://doi.org/10.1002/glia.22983 source

  • May combine two claims.

    Interpretationown data

    Exchange of miR-146a-5p (and possibly other miRNAs) between inflammatory microglia and neurons via EVs causes important alterations of synaptic structure and function, independent of neuronal damage.

    “The main accomplishment of our study is the demonstration that exchange of miR-146a-5p (and possibly other miRNAs) between inflammatory microglia and neurons via EVs causes important alterations of synaptic structure and function, independent of neuronal damage.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The inflammatory stimuli used in the study model chronic brain inflammation relevant to human neuroinflammatory disorders.

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

    Premisecitation

    Microglia can regulate synapses through contact-dependent mechanisms, including engulfment of synaptic elements.

    “Recent studies have revealed that microglia can also regulate synapses through contact-dependent mechanisms, including engulfment of synaptic elements”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [74] Tremblay ME, Stevens B, Sierra A, Wake H, Bessis A, Nimmerjahn A (2011) The role of microglia in the healthy brain. J Neurosci Off J Soc Neurosci 31:16064–16069. https://doi.org/10.1523/JNEUROSCI.4158-11.2011 source

      Not checked in the free run.

    • cites [80] Wake H, Moorhouse AJ, Jinno S, Kohsaka S, Nabekura J (2009) Resting microglia directly monitor the functional state of synapses in vivo and determine the fate of ischemic terminals. J Neurosci Off J Soc Neurosci 29:3974–3980. https://doi.org/10.1523/JNEUROSCI.4363-08.2009 source
    • cites [82] Wu Y, Dissing-Olesen L, MacVicar BA, Stevens B (2015) Microglia: dynamic mediators of synapse development and plasticity. Trends Immunol 36:605–613. https://doi.org/10.1016/j.it.2015.08.008 source

      Not checked in the free run.

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

    Premisecitation

    A quarter of MS patients suffer symptoms arising from neuronal dysfunction, such as episodic memory impairment, already at the first clinical episode.

    “Despite a quarter of MS patients suffer symptoms arising from neuronal dysfunction, such as episodic memory impairment, already at the first clinical episode and show hippocampal microstructural damage [57, 63] we still lack a full understanding of how synaptic alterations start and how they are linked to microglia activation.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [57] Planche V, Ruet A, Coupe P, Lamargue-Hamel D, Deloire M, Pereira B, Manjon JV, Munsch F, Moscufo N, Meier DS et al (2016) Hippocampal microstructural damage correlates with memory impairment in clinically isolated syndrome suggestive of multiple sclerosis. Mult Scler 23:1214. https://doi.org/10.1177/1352458516675750 source
    • cites [63] Rocca MA, Amato MP, De Stefano N, Enzinger C, Geurts JJ, Penner IK, Rovira A, Sumowski JF, Valsasina P, Filippi M et al (2015) Clinical and imaging assessment of cognitive dysfunction in multiple sclerosis. Lancet Neurol 14:302–317. https://doi.org/10.1016/S1474-4422(14)70250-9 source

  • May combine two claims.

    Interpretationcitation

    EV fusion allows direct retrieval of EV cargo in the cytosol of neurons, avoiding degradation in late endosome or lysosomes, where EVs may accumulate after endocytosis.

    “EV fusion allows direct retrieval of EV cargo in the cytosol of neurons, avoiding degradation in late endosome or lysosomes, where EVs may accumulate after endocytosis [13, 26, 28].”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [13] Chivet M, Javalet C, Laulagnier K, Blot B, Hemming FJ, Sadoul R (2014) Exosomes secreted by cortical neurons upon glutamatergic synapse activation specifically interact with neurons. J Extracell Vesicles 3:24722. https://doi.org/10.3402/jev.v3.24722 source
    • cites [26] Fitzner D, Schnaars M, van Rossum D, Krishnamoorthy G, Dibaj P, Bakhti M, Regen T, Hanisch UK, Simons M (2011) Selective transfer of exosomes from oligodendrocytes to microglia by macropinocytosis. J Cell Sci 124:447–458. https://doi.org/10.1242/jcs.074088 source
    • cites [28] Fruhbeis C, Frohlich D, Kuo WP, Amphornrat J, Thilemann S, Saab AS, Kirchhoff F, Mobius W, Goebbels S, Nave KA et al (2013) Neurotransmitter-triggered transfer of exosomes mediates oligodendrocyte-neuron communication. PLoS Biol 11:e1001604. https://doi.org/10.1371/journal.pbio.1001604 source
    • Rests on an assumption: The EV fusion mechanism observed in cultured cells occurs in vivo.
62 of 62

  • The paper relies on this without support.

    The mechanism(s) leading to microglia-mediated synapse loss during brain inflammation are unknown.

    “However, the mechanism(s) leading to microglia-mediated synapse loss during brain inflammation remain unknown”

  • The paper relies on this without support.

    Whether reactive microglia may disrupt synaptic structure and function by silencing synaptic genes has never been explored.

    “and whether reactive microglia may disrupt synaptic structure and function by silencing synaptic genes has never been explored.”

  • The paper relies on this without support.

    No formal proof for glia-to-neuron transfer of miRNAs has been provided.

    “Despite these advances, no formal proof for glia-to-neuron transfer of miRNAs has been provided”

  • The paper relies on this without support.

    Whether vesicular miRNA is essential to alter gene expression in recipient neurons remains to be established.

    “and whether vesicular miRNA is essential to alter gene expression in recipient neurons remains to be established.”

  • The paper relies on this without support.

    Nlg1 is an adhesion protein that plays a crucial role in dendritic spine formation and synaptic stability.

    “postsynaptic neuroligin1 (Nlg1), an adhesion protein which play a crucial role in dendritic spine formation and synaptic stability.”

  • The paper relies on this without support.

    cel-miR-39 is not expressed in rat cells.

    “a C. elegans miRNA, not expressed in rat cells”

  • The paper relies on this without support.

    cel-miR-39 cannot be upregulated in neurons upon contact with MVs.

    “which cannot be upregulated in neurons upon contact with MVs”

  • The paper relies on this without support.

    Annexin-V cloaks phosphatidyl-serine (PS) residues on MV surface.

    “Cloaking phosphatidyl-serine (PS) residues on MV surface with annexin-V decreased MV adhesion by 54%”

  • The paper relies on this without support.

    Heparitinase I cleaves HS chain of transmembrane proteoglycans (syndecans).

    “heparitinase I, which cleaves HS chain of transmembrane proteoglycans (syndecans)”

  • The paper relies on this without support.

    The inflammatory stimuli used in the study model chronic brain inflammation relevant to human neuroinflammatory disorders.

    “In this study, we have identified miRNAs as additional active components of inflammatory MVs that are responsible for a delayed destabilization of excitatory synapses and decreased transmission, which selectively occur during chronic inflammation.”

  • The paper relies on this without support.

    The synaptic alterations observed in rodent hippocampal cultures and CA1 translate to human MS cognitive symptoms.

    “This evidence provides a possible link between microglia activation, enhanced EV production and cognitive symptoms in MS patients.”

  • The paper relies on this without support.

    The miRNAs detected in CSF EVs from MS patients are derived from microglia and are functionally transferred to neurons.

    “Here we show that miR-146a-5p, together with miR-181a and miR-223 are present in EVs isolated from the CSF of a group of MS patients.”

  • The paper relies on this without support.

    The EV fusion mechanism observed in cultured cells occurs in vivo.

    “A prominent finding of our study is that glial EVs deliver their cargo to neurons by transient fusion.”

  • The paper relies on this without support.

    Cognitive symptoms in MS are caused by synaptic dysfunction or loss.

    “which may underlie synaptic dysfunction occurring in chronic inflammation.”
14 of 14

Claims other papers make about this one

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

Self-check

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

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

  • Conflicting synaptic target stated for miR-181a

    Minor

    The Results identify GluR2 as the target of miR-181a, whereas the Discussion says miR-181a regulates only gluN2B. These are different receptors.

  • Figure 3f names NR2B as measured but reports SNAP25 statistics

    Minor

    The caption lists NR2B among the proteins analysed in the coculture western blot, but its reported tests instead include SNAP25, with no NR2B result identified.

  • Abstract calls miR-146a-5p microglia-specific despite astrocyte expression

    Minor

    The abstract describes the miRNA as microglia-specific, but the Results report its detection in cultured astrocytes and their MVs.

3 of 3

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Authors
Prada, I., Gabrielli, M., Turola, E., Iorio, A., D’Arrigo, G., Parolisi, R., De Luca, M., Pacifici, M., Bastoni, M., Lombardi, M., Legname, G., Cojoc, D., Buffo, A., Furlan, R., Peruzzi, F., & Verderio, C.
Affiliations
  • University of Modena and Reggio Emilia (IT)
  • Scuola Internazionale Superiore di Studi Avanzati (IT)
  • Neuroscience Institute Cavalieri Ottolenghi (IT)
  • Louisiana State University Health Sciences Center New Orleans (US)
  • AREA Science Park (IT)
  • National Research Council (IT)
Corresponding author
Verderio, Claudia
ORCID
Venue
Acta Neuropathologica
Publisher
Springer Science and Business Media LLC
Published
2018-01-04
Volume
135
Issue
4
Pages
529-550
ISSN
  • 0001-6322 (ISSN-L)
  • 1432-0533
DOI
10.1007/s00401-017-1803-x
PMC
PMC5978931
OpenAlex
W2781721887
Cited by
312
References
88
Citations by year
2026: 19 · 2025: 30 · 2024: 57 · 2023: 34 · 2022: 42 · 2021: 63 · 2020: 25 · 2019: 28 · 2018: 13
FWCI
11.47
Type
journal article
Language
en
Open access
Licence
Locations
  • Acta Neuropathologica (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • SDL (submittedVersion, cc-by)
  • Institutional Research Information System University of Turin (University of Turin) (submittedVersion, cc-by)
  • Archivio Istituzionale della Ricerca - Università Vita-Salute San Raffaele (submittedVersion)
Funders
  • Fondazione Italiana Sclerosi Multipla (grant 2016/R/30)
  • ERA-NET NEURON JTC 2014 (Micromet project)
  • GMSI 2015 (GMSI 2015)
  • Fondazione Umberto Veronesi (2016)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
Keywords
extracellular vesicles, microRNA transfer, miR-146a, microglia, neuroinflammation, synaptic dysfunction, dendritic spine loss, excitatory synapses, multiple sclerosis, hippocampal neurons, neuron-microglia communication
MeSH terms
Extracellular Vesicles, Animals, Cells, Cultured, Cerebrospinal Fluid, Female, Hippocampus, Humans, Inflammation, Male, Mice, Inbred C57BL, Neuroglia, Neuronal Plasticity, Neurons, Synapses, Rats, Sprague-Dawley, Coculture Techniques, MicroRNAs, Primary Cell Culture

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 2026-11-06.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-11-06.

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.

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

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