CoChat
Sign Up
Cited by 255MicroRNA in disease regulationNeurogenesis and neuroplasticity mechanismsDevelopmental Biology and Gene Regulation

Comprehensive expression analyses of neural cell-type-specific miRNAs identify new determinants of the specification and maintenance of neuronal phenotypes

Open access
Jovičić, Ana, Roshan, Reema, Moisoi, Nicoleta, Pradervand, Sylvain, Moser, Roger, Pillai, Beena, Luthi-Carter, Ruth·2013·The Journal of Neuroscience
DOI 10.1523/jneurosci.0600-12.2013PMC PMC6705001
Open this paper in CoChat — free accountOpen PDF

Abstract

MicroRNAs (miRNAs) have been shown to play important roles in both brain development and the regulation of adult neural cell functions. However, a systematic analysis of brain miRNA functions has been hindered by a lack of comprehensive information regarding the distribution of miRNAs in neuronal versus glial cells. To address this issue, we performed microarray analyses of miRNA expression in the four principal cell types of the CNS (neurons, astrocytes, oligodendrocytes, and microglia) using primary cultures from postnatal d 1 rat cortex. These analyses revealed that neural miRNA expression is highly cell-type specific, with 116 of the 351 miRNAs examined being differentially expressed fivefold or more across the four cell types. We also demonstrate that individual neuron-enriched or neuron-diminished RNAs had a significant impact on the specification of neuronal phenotype: overexpression of the neuron-enriched miRNAs miR-376a and miR-434 increased the differentiation of neural stem cells into neurons, whereas the opposite effect was observed for the glia-enriched miRNAs miR-223, miR-146a, miR-19, and miR-32. In addition, glia-enriched miRNAs were shown to inhibit aberrant glial expression of neuronal proteins and phenotypes, as exemplified by miR-146a, which inhibited neuroligin 1-dependent synaptogenesis. This study identifies new nervous system functions of specific miRNAs, reveals the global extent to which the brain may use differential miRNA expression to regulate neural cell-type-specific phenotypes, and provides an important data resource that defines the compartmentalization of brain miRNAs across different cell types.

AI snapshot

Written from the paper’s full text.

TL;DR

This study profiled miRNAs across the four principal CNS cell types and found highly cell-type-specific expression, with 116 of 351 examined miRNAs differing at least fivefold. Functional tests showed neuron-enriched miRNAs promote neuronal differentiation while glia-enriched miRNAs inhibit it and can suppress inappropriate neuronal gene expression in glia.

Key findings

  • Microarray analysis of neurons, astrocytes, oligodendrocytes, and microglia from postnatal day 1 rat cortex found 116 of 351 miRNAs differentially expressed at least fivefold across the four cell types.
  • Restricting to neurons, astrocytes, and oligodendrocytes, the study identified 53 neuron-enriched and 23 neuron-diminished miRNAs, 13 astrocyte-enriched and 9 astrocyte-diminished miRNAs, and 13 oligodendrocyte-enriched and 4 oligodendrocyte-diminished miRNAs.
  • Overexpression of neuron-enriched miR-376a and miR-434 increased neural stem cell differentiation into neurons, whereas glia-enriched miR-223, miR-146a, miR-19, and miR-32 decreased neuronal differentiation.
  • Glia-enriched miR-146a targeted neuron-specific genes including Nlgn1, Nova1, and Syt1; miR-146a inhibited neuroligin 1-dependent synaptogenesis, suggesting glial miRNAs prevent inappropriate neuronal protein expression and phenotypes.

Methods

  • Primary cultures from postnatal day 1 rat cortex were used for neurons, astrocytes, oligodendrocytes, and microglia; miRNA expression was profiled on Agilent rat miRNA microarrays with three biological replicates for neurons, oligodendrocytes, and microglia and four for astrocytes.
  • Differential expression was analyzed with limma using an F test across pairwise cell-type contrasts, with FDR <1%, at least fivefold change versus the average of other groups, at least twofold change versus each other group individually, and presence filters.
  • Functional validation used lentiviral miRNA overexpression in rat cortical stem cells, LNA antagomirs in PC12 and C6 cells, 3′-UTR luciferase reporter assays, immunolabeling, immunoblotting, FACS of Thy1-EGFP and Gfap-EGFP mouse brain cells, and a neuroligin 1-dependent synapse formation assay.

Limitations

  • The paper states that cell cultures derived from neonatal brain may still be developing and therefore may not represent mature in vivo phenotypes; it addressed this by validating selected miRNAs in acutely isolated mouse brain cells.
  • The paper states that high miRNA overexpression sometimes decreased stem cell numbers, so vector concentration was reduced and data from treatments decreasing cell numbers to <35% of noninfected control were excluded.
  • Evident from the text: the microarray covered 351 miRNAs, in vivo validation was limited to selected miRNAs, and for Syt1 the predicted miR-146a binding-site mutation did not abolish regulation, leaving direct versus indirect targeting unresolved.

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

Integrity signals

Registry

Crossref · 10.1523/jneurosci.0600-12.2013

Cites

53 of 53 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

4 of 43 cited sources checked · 1 supported · 0 flagged · accuracy not yet measured

  • Findingown data

    We confirmed the glial expression of miR-219 and miR-338.

    “Similarly, we confirmed the glial expression of miR-219 and miR-338, which have been implicated previously in the differentiation of oligodendrocytes”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites Lau et al., 2008 source
    • cites Dugas et al., 2010

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

    • cites Zhao et al., 2010

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

  • Premisecitation

    Negative regulation of Hes5 promotes neuronal differentiation.

    “the negative regulation of which has also been found to promote neuronal differentiation in other studies (Ohtsuka et al., 1999; Fig. 7 I, K ).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites Ohtsuka et al., 1999 source

      The indexed Abstract reports that activated Notch inhibited neuronal differentiation in Hes5-null cells, but not in cells lacking both Hes1 and Hes5. This supports a role for Hes5 alongside Hes1, not the claim that negative regulation of Hes5 alone promotes neuronal differentiation. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
  • Findingown data

    Several miRNAs had strong effects on neural stem cell differentiation.

    “These results show that several miRNAs had strong effects on neural stem cell differentiation.”
    Quote in the paperSubstantial claim
  • Findingown data

    Three miRNA-miRNA* pairs had opposite expression patterns, maintaining a different strand in microglia compared with the other three cell types: miR-125-5p/miR-125-3p, miR-138/miR-138*, and miR-99b/miR-99b*.

    “Interestingly, however, there were three miRNA-miRNA* pairs that were found to have opposite expression patterns (i.e., they maintained a different strand in microglia compared with the other three cell types).These pairs are as follows: miR-125-5p/ miR-125-3p, miR-138/miR-138*, and miR-99b/miR-99b* (Fig. 5).”
    Quote in the paperSubstantial claim
  • Findingown data

    Overexpression of the neuron-enriched miRNAs miR-376a and miR-434 increases the differentiation of neural stem cells into neurons.

    “overexpression of the neuron-enriched miRNAs miR-376a and miR-434 increased the differentiation of neural stem cells into neurons”
    • Also stated in: § Genomic colocalization and coexpression of neural cell-typespecific miRNAs · plain
      “Of our newly identified neuron-enriched miRNAs, miR-376a and miR-434 also increased the number of cells differentiating toward a neuronal phenotype, with an effect size similar to miR-124 (Fig. 6C).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: miRNAs bind to mRNAs by sequence complementarity and direct their degradation or translational repression.
  • May combine two claims. 20
  • Classified as probably not a substantial claim. 20
  • The quote only partly states this claim. 1

  • May combine two claims.

    Premisecitation

    miRNAs play important roles in both brain development and the regulation of adult neural cell functions.

    “MicroRNAs (miRNAs) have been shown to play important roles in both brain development and the regulation of adult neural cell functions.”
    • Also stated in: § Introduction · plain
      “Recent studies have shown that numerous miRNAs exist in mammalian systems, where they play important roles in development, are responsible for regulating cell-type-specific functions in the adult organism, and are involved in disease processes (Bartel, 2009).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites Bartel, 2009 source

      In “Roles of specific miRNA:target interactions” (page not provided), the review says miRNA regulation is needed for development of the Drosophila peripheral nervous system, but does not establish a role in brain development specifically. “Binary switches and much more” describes miR-8 regulation that prevents neurodegeneration, but does not identify this as regulation of adult neural cell functions. (LLM, conf=0.84)

      Experimental — accuracy not yet measured

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

    Premisecitation

    miR-124 is a potent determinant of neuronal identity.

    “While confirming that miR-124 is a potent determinant of neuronal identity (Krichevsky et al., 2003;Visvanathan et al., 2007)”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites Krichevsky et al., 2003 source

      The ‘miRNA expression during corticogenesis’ section reports that miR-124a expression rose more than 13-fold during embryonic brain development. That supports an association with development, but not the claim that miR-124 determines neuronal identity; the Introduction says that no mammalian miRNA function had yet been demonstrated. Page numbers are unavailable in our copy. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites Visvanathan et al., 2007

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

  • May combine two claims.

    Findingown data

    This analysis yielded multiple enriched or diminished miRNAs for neurons and microglia and relatively fewer miRNAs detected as uniquely enriched or diminished in astrocytes or oligodendrocytes.

    “This analysis yielded multiple enriched or diminished miRNAs for neurons and microglia and relatively fewer miRNAs detected as uniquely enriched or diminished in astrocytes or oligodendrocytes (Table 1).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Neural miRNA expression is highly cell-type specific, with 116 of the 351 miRNAs examined being differentially expressed fivefold or more across the four cell types.

    “These analyses revealed that neural miRNA expression is highly cell-type specific, with 116 of the 351 miRNAs examined being differentially expressed fivefold or more across the four cell types.”
    • Also stated in: § Individual neural cell types have distinct miRNA expression profiles · plain
      “These analyses showed that neural cell types have remarkably distinct miRNA expression profiles (Fig. 2), with 116 of the examined 351 miRNAs being at least fivefold differentially expressed across the four cell types.”
    • Also stated in: § Introduction · plain
      “Consistent with our hypothesis, our analyses showed that neural cell subtypes differed extensively in their miRNA expression patterns.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Among miRNAs exhibiting cell-type-specific expression among the four cell types, only 3 clusters were detected: a microglia-enriched cluster on Chr4q42 comprising miR-141 and miR-200c and two neuron-enriched clusters on Chr6q32.

    “Among the miRNAs exhibiting cell-type-specific expression among the four cell types, only 3 clusters were detected: a microglia-enriched cluster on Chr4q42 comprising miR-141 and miR-200c and two neuron-enriched clusters on Chr6q32”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Restricting analyses to neurons, astrocytes, and oligodendrocytes identified 53 enriched and 23 diminished miRNAs in neurons, 13 enriched and 9 diminished miRNAs in astrocytes, and 13 enriched and 4 diminished miRNAs in oligodendrocytes.

    “we identified 53 enriched and 23 diminished miRNAs in neurons, 13 enriched and 9 diminished miRNAs in astrocytes, and 13 enriched and 4 diminished miRNAs in oligodendrocytes (Table 2).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The astrocyte-enriched expression of miR-143, miR-193, and miR-449a and astrocyte-diminished expression of miR-327 and miR-494 were confirmed in Gfap-EGFP mouse brain cells.

    “By implementing the same approach using Gfap-EGFP transgenic mice, we likewise confirmed the astrocyte-enriched expression of miR-143, miR-193, and miR-449a and the astrocyte-diminished expression of miR-327 and miR-494 (Fig. 3C,D).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingcitation

    Neuronal miR-124 increased the differentiation of stem cells toward neuronal cell fate, in agreement with previous observations.

    “As a positive control, we showed that neuronal miR-124 increased the differentiation of stem cells toward neuronal cell fate, in agreement with previous observations (Krichevsky et al., 2003).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Krichevsky et al., 2003 source

      Queued — the background check reaches it, usually within a day.

  • May combine two claims. The quote only partly states this claim.

    Findingown data

    Of 43 duplex pairs identified among differentially expressed miRNAs, 39 showed correlated expression in which one of the two strands was always present at higher levels, but the relative expression of the two strands across neural cell types retained the same profile.

    “Of 43 duplex pairs we identified among our differentially expressed miRNAs, 39 showed correlated expression in which one of the two strands was always present at higher levels”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Comparing differential miRNA expression only across neurons, astrocytes, and oligodendrocytes, four additional clusters were found: two neuron-diminished clusters on Chr15q24 and ChrXq36, and two astrocyte-enriched clusters on ChrXq12 and Chr8q24.

    “Comparing differential miRNA expression only across neurons, astrocytes, and oligodendrocytes, four additional clusters were found: two neuron-diminished clusters on Chr15q24 comprisingmiR-17-3p,miR-17-5p,miR-18a,miR-19a,miR-20a,miR-20a*, and miR-92a and on ChrXq36 comprising miR-322, miR-322*, miR-351, miR-450a, miR-503, miR-542-3p, miR-542-5p, and two astrocyte-enriched clusters on ChrXq12”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Hierarchical clustering showed that cell types of common developmental origin differed least in their miRNA composition (astrocytes and oligodendrocytes), while neurons and microglia showed more unique miRNA expression profiles.

    “Moreover, hierarchical clustering of the expression profiles showed that cell types of common developmental origin differed least in their miRNA composition (astrocytes and oligodendrocytes), while neurons and microglia showed more unique miRNA expression profiles.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductreasoning

    The concentration of the applied vector was decreased to limit overexpression levels of each miRNA to a maximum of 100-fold compared with endogenous.

    “Therefore, we decreased the concentration of the applied vector to limit overexpression levels of each miRNA to a maximum of 100-fold compared with endogenous.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S2 (checkable).

    Premisereasoning

    The full scope of miRNA-mediated regulation of brain functions is largely unknown.

    “However, the full scope of miRNA-mediated regulation of brain functions is largely unknown.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Study design and conductown data

    FACS on brain samples of mice expressing EGFP under the Thy1 promoter separated neurons from glial cells, as confirmed by enrichment of neuronal marker Tubb3 and neuronal miR-124 in Thy1+ cells compared with Thy1- cells.

    “By using FACS on brain samples of mice expressing EGFP under the control of the Thy1 promoter, we were able to successfully separate neurons from glial cells as confirmed by enrichment of neuronal marker Tubb3 and neuronal miR-124 in Thy1 ϩ cells compared with Thy1 Ϫ cells (Fig. 3A).”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Brain parenchyma is principally composed of the four neural cell types (neurons, astrocytes, oligodendrocytes, and microglia), with insignificant contributions from endothelial cells, pericytes, fibroblasts, smooth muscle cells, and erythrocytes.

    “As we have shown in recent cell-resolved analyses of brain tissue, brain parenchyma is principally composed of the 4 neural cell types (neurons, astrocytes, oligodendrocytes, and microglia), with insignificant contributions from endothelial cells, pericytes, fibroblasts, smooth muscle cells, and erythrocytes (Kuhn et al., 2011).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Kuhn et al., 2011 source

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

    Study design and conductnone given

    Data from treatments that decreased the numbers of cells to less than 35% of the noninfected control were not considered.

    “In addition, we did not take into consideration any data from treatments that decreased the numbers of cells to Ͻ35% of the noninfected control.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    miRNAs show varying levels in different organs, consistent with their anticipated role in regulating tissue-specific protein expression.

    “Interestingly, miRNAs show varying levels in different organs, which is consistent with their anticipated role in regulating tissue-specific protein expression (Lagos-Quintana et al., 2002).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Lagos-Quintana et al., 2002

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

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

    Premisecitation

    Previous studies had compared miRNA expression in whole brain tissue with other tissues or assessed differences during development of individual neural cell types.

    “Previous studies had instead compared the miRNA expression in whole brain tissue with the expression in other tissues of the same organism (Sempere et al., 2004;Wienholds et al., 2005) or assessed differences in miRNA expression during different stages of development of individual neural cell types (Krichevsky et al., 2003;Smirnova et al., 2005;Zhao et al., 2010).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites Sempere et al., 2004 source

      Queued — the background check reaches it, usually within a day.

    • cites Wienholds et al., 2005 source
    • cites Krichevsky et al., 2003 source

      Queued — the background check reaches it, usually within a day.

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

    Premisecitation

    The largest miRNA cluster discovered to date consists of more than 50 miRNAs in the human imprinted 14q32 domain, with orthologous clusters in rat (6q32) and mouse (12qF1).

    “The largest miRNA cluster discovered to date consists of Ͼ50 miRNAs in the human imprinted 14q32 domain (Seitz et al., 2004), which has orthologous clusters in the rat (6q32) and mouse (12qF1).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Seitz et al., 2004 source

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

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

    Premisecitation

    A few miRNAs within the 14q32 cluster have been implicated in regulating synapse-specific genes and shown to exhibit neuronal-activity-dependent expression.

    “Interestingly, a few miRNAs within this cluster have been implicated in regulating synapse-specific genes (Schratt et al., 2006) and shown to exhibit neuronal-activitydependent expression (Fiore et al., 2009).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Schratt et al., 2006 source
    • cites Fiore et al., 2009 source

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

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

    Premisecitation

    The strand in which the 5' end is less tightly paired to its complement remains associated with Ago and performs the functions of the mature miRNA.

    “Studies investigating the relative thermodynamic stability of the two ends of the pre-miRNA duplex indicated that the strand in which the 5Ј end is less tightly paired to its complement remains associated with Ago and performs the functions of the mature miRNA (Khvorova et al., 2003;Schwarz et al., 2003).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Khvorova et al., 2003 source
    • cites Schwarz et al., 2003 source

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

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

    Premisecitation

    The physiological importance of miRNAs in nervous system functions and disease states has been suggested by previous studies of a small number of brain-enriched miRNAs.

    “The physiological importance of miRNAs in nervous system functions and disease states has also been suggested by previous studies of a small number of brainenriched miRNAs (Leucht et al., 2008;Mellios et al., 2008;Packer et al., 2008;Cheng et al., 2009;Schratt, 2009).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Leucht et al., 2008 source
    • cites Mellios et al., 2008 source

      Queued — the background check reaches it, usually within a day.

    • cites Packer et al., 2008 source

      Queued — the background check reaches it, usually within a day.

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

    Premisecitation

    miRNAs are frequently found in genomic clusters.

    “miRNAs are frequently found in genomic clusters (Bartel, 2009).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Bartel, 2009 source

      Not checked in the free run.

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

    Premisecitation

    The brain has a particularly high percentage of tissue-specific and tissue-enriched miRNAs.

    “Compared with other organs, the brain has a particularly high percentage of tissue-specific and tissue-enriched miR-NAs (Lagos-Quintana et al., 2002;Kim et al., 2004;Sempere et al., 2004;Smirnova et al., 2005).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Lagos-Quintana et al., 2002

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

    • cites Kim et al., 2004

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

    • cites Sempere et al., 2004 source

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

  • May combine two claims.

    Study design and conductown data

    The primary cultures used for expression analyses represented the intended cell populations, as confirmed by immunocytochemistry and largely segregated expression of cell-type-specific mRNAs.

    “The cellular composition of these cultures was verified by immunocytochemistry using antibodies against well established cell-type-specific marker antigens: NeuN/Rbfox (neuronal nuclear antigen/RNA binding protein Fox) for mature neurons, GFAP (glial fibrillary acidic protein) for astrocytes, O4 antigen for oligodendrocytes, and integrin ␣ M chain (Itgam) for microglia (Kuhn et al., 2011).”
    • Also stated in: § Individual neural cell types have distinct miRNA expression profiles · hedged
      “These mRNA expression values indicated largely segregated expression of the cell-type-specific RNAs, further confirming that the primary cultures used for expression analyses represented the intended cell populations (Fig. 1).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Kuhn et al., 2011 source

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

    Premisecitation

    Neural tissue is highly heterogeneous, composed of different types of neurons, astrocytes, and oligodendrocytes, which develop from a common pool of neural progenitor cells, and microglia, which develop from the hematopoietic lineage.

    “Neural tissue is highly heterogeneous, being composed of different types of neurons, astrocytes, and oligodendrocytes, which de-velop from a common pool of neural progenitor cells (Gage, 2000), and microglia, which develop from the hematopoietic lineage (Ritter et al., 2006).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Gage, 2000 source
    • cites Ritter et al., 2006 source

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

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

    Premisecitation

    Strand selection by Ago is not symmetrical.

    “The small RNA cloning strategy used to identify mature miRNAs revealed that the process of the strand selection by Ago is not symmetrical (Lim et al., 2003).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Lim et al., 2003 source

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

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

    Premisecitation

    miRNAs are 19-24 nucleotide noncoding RNAs that act as important regulators of posttranscriptional gene expression.

    “MicroRNAs (miRNAs) are 19-to 24-nucleotide (nt) noncoding RNAs that act as important regulators of posttranscriptional gene expression (Ambros, 2004;Kim, 2005).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Ambros, 2004 source
    • cites Kim, 2005 source

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

    Premisecitation

    Mef2c and Neurog1 stimulate neuronal differentiation.

    “which have been reported to stimulate neuronal differentiation (Fode et al., 2000;Li et al., 2008; Fig. 7 F, H, J, L).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Fode et al., 2000 source

      Not checked in the free run.

    • cites Li et al., 2008

      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).

    Interpretationreasoning

    Targeting of other neuronal mRNAs by miR-146a and other glia-enriched miRNAs might likewise prevent glia from mistakenly adopting neuron-specific phenotypes.

    “By extension, this also implies that the targeting of other neuronal mRNAs by miR-146a and other glia-enriched miRNAs might likewise prevent glia from mistakenly adopting neuron-specific phenotypes.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Heterologous expression of key synaptogenic proteins in non-neuronal cells allows them to receive projections from neurons and subsequently induce morphological and functional presynaptic differentiation in the contacting axons.

    “It has been shown previously that heterologous expression of key synaptogenic proteins in non-neuronal cells allows them to receive projections from neurons and subsequently induce morphological and functional presynaptic differentiation in the contacting axons (Biederer and Scheiffele, 2007).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Biederer and Scheiffele, 2007 source

  • May combine two claims.

    Interpretationreasoning

    Cell-type-specific miRNAs have important roles in both establishing and maintaining distinct neuronal and glial phenotypes.

    “These studies therefore provide novel evidence that cell-type-specific miRNAs have important roles in both establishing and maintaining distinct neuronal and glial phenotypes.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Targeting of Stat3 mediates miR-124-dependent proneuronal differentiation of neural stem cells.

    “the targeting of which has been shown previously to mediate miR-124-dependent proneuronal differentiation of neural stem cells (Krichevsky et al., 2003;Fig. 7 E, G).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Krichevsky et al., 2003 source

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

  • May combine two claims.

    Interpretationown data

    Syt1 is a target of miR-146a, but the actual binding site is different from the predicted one.

    “this result implies that Syt1 is a target of miR-146a, but that the actual binding site is different from the predicted one”
    Quote in the paperSubstantial claim
34 of 34

  • The paper relies on this without support.

    miRNAs bind to mRNAs by sequence complementarity and direct their degradation or translational repression.

    “miRNAs bind to messenger RNAs (mRNAs) based on sequence complementarity and direct the degradation or repression of translation of the mRNAs to which they are bound (known as their targets).”

  • The paper relies on this without support.

    The heterogeneous phenotypes of neural cells must be established during cell specification and maintained throughout the life of the adult organism.

    “The heterogeneous phenotypes of the various neural cells must be established during cell specification and be maintained throughout the life of the adult organism.”

  • The paper relies on this without support.

    A genome-wide investigation of miRNA expression in neurons, astrocytes, oligodendrocytes, and microglia had not been available previously.

    “However, a genome-wide investigation of miRNA expression in neurons, astrocytes, oligodendrocytes, and microglia had not been available previously.”

  • The paper relies on this without support.

    Sequence-based target prediction alone is subject to false-positive identification.

    “Because sequence-based target prediction alone is subject to false-positive identification”
4 of 4

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?

  • miR-146a-5p is highly enriched in microglia compared to hippocampal neurons and astrocytes.

    “A previous study reported that miR-146a-5p is a microglia-enriched miRNA, not present in hippocampal neurons, and expressed at much lower extent in astrocytes.29”

    From Microglia secrete miR-146a-5p-containing exosomes to regulate neurogenesis in depression (2022) · cites this paper as [29]

    Table 1 (section ‘Individual neural cell types have distinct miRNA expression profiles’; page unavailable) lists miR-146a as enriched in microglia and diminished in neurons relative to the other cell types, supporting the enrichment comparison. It does not establish that miR-146a is absent from hippocampal neurons: ‘diminished’ is not ‘not present,’ and the table does not specify hippocampal neurons. Its enrichment criteria also support higher expression in microglia than astrocytes. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • 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].”

    From Glia-to-neuron transfer of miRNAs via extracellular vesicles: a new mechanism underlying inflammation-induced synaptic alterations (2018) · cites this paper as [39]

    The section “Glia-enriched miR-146a targets neuron-specific genes and functions” reports that miR-146a represses neuron-specific targets and helps prevent glial cells from adopting neuronal features. But its experiments concern glial cells, including primary astrocytes, not neuronal progenitors. The section “Individual neural cell types have distinct miRNA expression profiles” lists miR-146a as astrocyte-enriched relative to neurons and oligodendrocytes; the retrieved text does not establish the claimed greater enrichment in microglia. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • miRs often display cell type-specific regulation of their targets.

    “Additionally, miRs often display cell type–specific regulation of their targets. 62 , 63”

    From Deletion of miR-33, a regulator of the ABCA1–APOE pathway, ameliorates neuropathological phenotypes in APP/PS1 mice (2024) · cites this paper as [62]

  • Previous studies identified a set of miRNAs enriched in neurons, with expression fivefold higher than glial cells, including miR-124, miR-132 and miR-135b, while the same cells display very low expression of miR-21 and miR-146a compared to glial cells.

    “This identified a set of miRNAs that are enriched in neurons, with expression fivefold higher than the glial cells, including miR-124, miR-132 and miR-135b, while the same cells display very low expression of miR-21 and miR-146a compared to glial cells [8].”

    From Shared and distinct microRNA profiles between HT22, N2A and SH-SY5Y cell lines and primary mouse hippocampal neurons (2025) · cites this paper as [8]

  • Glial-enriched miR-146a-5p can inhibit neuronal differentiation.

    “It has been reported that glial-enriched miR-146a-5p can inhibit neuronal differentiation.29”

    From Microglia secrete miR-146a-5p-containing exosomes to regulate neurogenesis in depression (2022) · cites this paper as [29]

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.

  • Tested neuron-enriched miRNA changes from miR-367a to miR-376a

    Minor

    The list of neuron-enriched miRNAs selected for overexpression names miR-367a, but the subsequent account of the results names miR-376a instead.

  • Figure 7B compares C6 cells with themselves

    Minor

    Figure 7B says neuronal miRNAs are enriched in C6 cells compared with C6 cells. Its own stated statistical comparison is between C6 and PC12 samples, not two C6 groups.

  • Figure 4 duplicates miR-20a in the Chr15q24 cluster

    Minor

    The Results list miR-20a and miR-20a* as distinct members of this cluster, whereas the Figure 4 caption lists miR-20a twice and omits miR-20a*.

  • Chromosome locations of two astrocyte-enriched clusters are reversed

    Minor

    The Results place miR-34b/miR-34c on ChrXq12 and miR-221/miR-222 on Chr8q24. The Figure 4 caption assigns each pair to the other chromosome location.

4 of 4

Cites

Flagged first, then by citations

Cited by

Sorted by citations

Related

Authors
Jovičić, A., Roshan, R., Moisoi, N., Pradervand, S., Moser, R., Pillai, B., & Luthi-Carter, R.
Affiliations
  • École Polytechnique Fédérale de Lausanne (CH)
  • Institute of Genomics and Integrative Biology (IN)
  • University of Leicester (GB)
  • University of Lausanne (CH)
ORCID
Venue
The Journal of Neuroscience
Publisher
Society for Neuroscience
Published
2013-03-20
Volume
33
Issue
12
Pages
5127-5137
ISSN
  • 0270-6474 (ISSN-L)
  • 1529-2401
DOI
10.1523/jneurosci.0600-12.2013
PMC
PMC6705001
OpenAlex
W2020613231
Cited by
251
References
53
Citations by year
2026: 4 · 2025: 11 · 2024: 13 · 2023: 18 · 2022: 13 · 2021: 29 · 2020: 24 · 2019: 24 · 2018: 20 · 2017: 23 · 2016: 22 · 2015: 20 · 2014: 18 · 2013: 12
FWCI
8.43
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Neuroscience (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • SERVAL (Université de Lausanne) (publishedVersion, other-oa)
  • IRIS (submittedVersion)
Funders
Council of Scientific and Industrial Research, India (BSC 0123)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
  • Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
Keywords
microRNA, miR-146a, miR-376a, miR-223, neural stem cell differentiation, astrocytes, oligodendrocytes, microglia, neuroligin-1, synaptogenesis, glial reprogramming
MeSH terms
Animals, Animals, Newborn, Astrocytes, Cell Differentiation, Cerebral Cortex, Female, Male, Neuroglia, Neurons, Oligodendroglia, Phenotype, Synapses, Microglia, MicroRNAs, Rats, Neural Stem Cells, Transcriptome, 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 ›

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