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Cited by 116MicroRNA in disease regulationNeuroscience and Neuropharmacology ResearchMemory and Neural Mechanisms

The Brain-Enriched MicroRNA miR-9-3p Regulates Synaptic Plasticity and Memory

Open access
Sim, Su-Eon, Lim, Chae-Seok, Kim, Jae-Ick, Seo, Daekwan, Chun, Heejung, Yu, Nam-Kyung, Lee, Jaehyun, Kang, SukJae Joshua +11 more·2016·The Journal of Neuroscience
DOI 10.1523/jneurosci.0630-16.2016PMC PMC6601897
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Abstract

UNLABELLED: MicroRNAs (miRNAs) are small, noncoding RNAs that posttranscriptionally regulate gene expression in many tissues. Although a number of brain-enriched miRNAs have been identified, only a few specific miRNAs have been revealed as critical regulators of synaptic plasticity, learning, and memory. miR-9-5p/3p are brain-enriched miRNAs known to regulate development and their changes have been implicated in several neurological disorders, yet their role in mature neurons in mice is largely unknown. Here, we report that inhibition of miR-9-3p, but not miR-9-5p, impaired hippocampal long-term potentiation (LTP) without affecting basal synaptic transmission. Moreover, inhibition of miR-9-3p in the hippocampus resulted in learning and memory deficits. Furthermore, miR-9-3p inhibition increased the expression of the LTP-related genes Dmd and SAP97, the expression levels of which are negatively correlated with LTP. These results suggest that miR-9-3p-mediated gene regulation plays important roles in synaptic plasticity and hippocampus-dependent memory. SIGNIFICANCE STATEMENT: Despite the abundant expression of the brain-specific microRNA miR-9-5p/3p in both proliferating and postmitotic neurons, most functional studies have focused on their role in neuronal development. Here, we examined the role of miR-9-5p/3p in adult brain and found that miR-9-3p, but not miR-9-5p, has a critical role in hippocampal synaptic plasticity and memory. Moreover, we identified in vivo binding targets of miR-9-3p that are involved in the regulation of long-term potentiation. Our study provides the very first evidence for the critical role of miR-9-3p in synaptic plasticity and memory in the adult mouse.

AI snapshot

Written from the paper’s full text.

TL;DR

Inhibition of the brain-enriched microRNA miR-9-3p, but not miR-9-5p, impairs hippocampal long-term potentiation (LTP) and hippocampus-dependent memory in mice. miR-9-3p acts at least partly by suppressing LTP-related genes Dmd and SAP97, whose increased expression correlates with impaired LTP.

Key findings

  • miR-9-3p inhibition reduced theta-burst LTP at SC-CA1 synapses (last 5 min EPSC amplitude: control n=6 cells/3 mice; miR-9-3p n=9 cells/6 mice; naive n=7 cells/5 mice; one-way ANOVA F(2,19)=9.514, p=0.0014; Tukey control vs miR-9-3p and miR-9-3p vs naive, p<0.01), whereas miR-9-5p inhibition did not affect LTP (control n=12 cells/7 mice; miR-9-5p n=14 cells/6 mice; t(24)=0.8454, p=0.4062).
  • miR-9-3p inhibition did not significantly affect LTD (t(14)=0.6813, p=0.5068), basal synaptic transmission, membrane excitability, sEPSCs/sIPSCs/mEPSCs/mIPSCs, or NMDA receptor-mediated EPSCs.
  • miR-9-3p sponge-expressing mice showed impaired hippocampus-dependent memory: Morris water maze probe test (sponge × quadrant F(3,68)=4.44, p=0.0066; target quadrant control vs miR-9-3p p<0.05), object location memory (control t(7)=4.069, p=0.0048; miR-9-3p t(8)=0.9769, p=0.3572), and trace fear conditioning (last 4 test sessions t(6)=2.998, p=0.0241), without altered anxiety or locomotor activity.
  • miR-9-3p inhibition increased hippocampal protein levels of Dmd (control 100.0±14.0%, n=7 hippocampi; miR-9-3p 156.2±14.9%, n=8; t(13)=2.724, p=0.0173) and SAP97 (control 100.0±1.2%, n=5; miR-9-3p 125.2±7.8%, n=4; t(7)=3.606, p=0.0087), while miR-9-5p inhibition did not alter either.

Methods

  • Male C57BL/6N mice were used; AAV vectors expressing EGFP-tagged miR-9-3p, miR-9-5p, or control sponges under the human synapsin promoter were stereotaxically injected into dorsal CA1.
  • Electrophysiology used whole-cell patch-clamp recordings in acute hippocampal slices from 4- to 5-week-old mice; behavior used 8-week-old mice in Morris water maze, object location memory, trace fear conditioning, open-field, and elevated zero maze tasks.
  • Molecular validation combined TargetScan/Ingenuity Pathway Analysis target prediction, luciferase reporter assays with wild-type or mutated 3′ UTRs, and Western blots from CA1 regions expressing EGFP.

Limitations

  • The paper states that LTP was induced by theta-burst stimulation with relatively short intervals, which may not be sensitive to protein synthesis and PKA inhibitors; whether miR-9-3p also regulates the protein-synthesis-dependent form of LTP remains to be determined.
  • The paper notes that SAP97 was increased by miR-9-3p inhibition but no changes in excitatory synaptic transmission were detected, consistent with prior work in mature neurons.
  • Evident from the text: all experiments used male mice, so sex differences are not addressed; in vivo target validation focused on Dmd and SAP97 among seven candidate genes, leaving the other candidates unvalidated in vivo.

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

Integrity signals

Registry

Crossref · 10.1523/jneurosci.0630-16.2016

Cites

67 of 67 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 31 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Object location memory was significantly impaired in the miR-9-3p sponge group.

    “Object location memory was significantly impaired in the miR-9-3p sponge group”
    Quote in the paperSubstantial claim
  • Findingown data

    The miR-9-3p sponge specifically suppresses the activity of miR-9-3p.

    “Luciferase assay showing that the miR-9-3p sponge specifically suppresses the activity of miR-9-3p”
    Quote in the paperSubstantial claim
  • Findingown data

    miR-9-3p suppresses SAP97 expression.

    “Finally, we showed that miR-9-3p, but not miR-9-5p, suppresses the expression of the well known LTP-related genes Dmd and SAP97.”
    • Also stated in: § Discussion · plain
      “The SAP97 protein level was significantly increased in hippocampi overexpressing the miR-9-3p sponge”
    Quote in the paperSubstantial claim
  • Findingown data

    miR-9-3p is critical for LTP induction.

    “confirmed that miR-9-3p is critical for LTP, but not for LTD induction.”
    • Also stated in: § Discussion · plain
      “Theta-burst LTP recordings were significantly impaired in miR-9-3psponge-expressingneurons.”
    Quote in the paperSubstantial claim
  • Findingown data

    AAV-miR-9-3p sponge-expressing mice showed impaired spatial memory in the Morris water maze test.

    “AAV-miR-9-3p sponge-expressing mice showed impaired spatial memory in the Morris water maze test”
    Quote in the paperSubstantial claim
  • The quote only partly states this claim. 1
  • May combine two claims. 7
  • Classified as probably not a substantial claim. 13

  • The quote only partly states this claim.

    Findingown data

    Inhibition of miR-9-3p in vivo impairs hippocampus-dependent memory in multiple behavioral tasks.

    “Inhibition of miR-9-3p activity impairs hippocampus-dependent memories.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Basal anxiety level and locomotor activity were intact in both AAV-sponge groups.

    “Basal anxiety level and locomotor activity were intact in both AAV-sponge groups.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    miR-9-3p is involved in LTP induction, very likely through regulating AMPAR trafficking.

    “suggesting that miR-9-3p is involved in LTP induction, very likely through regulating AMPAR trafficking.”
    Quote in the paperSubstantial claim

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

    Premisenone given

    The role of SAP97 in excitatory synaptic transmission and AMPAR trafficking is controversial.

    “however, its role in excitatory synaptic transmission and AMPAR trafficking is controversial.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Dmd and SAP97 are novel targets of miR-9-3p.

    “Dmd and SAP97 are novel targets of miR-9-3p.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Inhibition of miR-9-5p impairs dendritic growth and excitatory synaptic transmission.

    “Recently, Giusti et al. (2014) showed that inhibition of miR-9-5p using sponge technique impairs dendritic growth and excitatory synaptic transmission.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Giusti et al., 2014 source

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

    Premisecitation

    Overexpressing SAP97 has no effect on synaptic transmission.

    “whereas others have shown that overexpressing SAP97 has no effect (Schnell et al., 2002;Schlu ¨ter et al., 2006).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Schnell et al., 2002 source
    • cites Schlüter et al., 2006

      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).

    Premisecitation

    miR-9 levels are decreased in postmortem brains of Huntington's disease patients.

    “the miR-9 level was shown to be decreased in the postmortem brains of Huntington's disease patients (Packer et al., 2008)”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • 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

    Expression levels of Dmd and SAP97 are negatively correlated with LTP level.

    “It is interesting that the expression levels of Dmd and SAP97 have been shown to be negatively correlated with the LTP level.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Vaillend et al., 2004 source
    • cites Nakagawa et al., 2004 source
    • cites Li et al., 2011

      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).

    Interpretationcitation

    miR-9-3p is critical for neuronal differentiation.

    “suggesting that miR-9-3p is critical for neuronal differentiation.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    miR-9-5p and miR-9-3p are abundantly expressed in postmitotic neurons.

    “However, miR-9-5p and miR-9-3p are abundantly expressed, not only in neural progenitors, but also in postmitotic neurons (Yuva-Aydemir et al., 2011;Liu et al., 2012).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Yuva-Aydemir et al., 2011 source

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

    • cites Liu et al., 2012 source

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

  • Classified as probably not a substantial claim.

    Premisecitation

    The induction of PKA-sensitive LTP requires multiple episodes of stimuli with proper interepisode intervals.

    “The induction of PKA-sensitive LTP requires multiple episodes of stimuli with proper interepisode intervals (Woo et al., 2003;Kim et al., 2010;Park et al., 2014;Park et al., 2016).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites Woo et al., 2003 source
    • cites Kim et al., 2010 source

      Not checked in the free run.

    • cites Park et al., 2014

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

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

    Interpretationown data

    Downregulation of Dmd by miR-9-3p may provide new insights for the development of treatments for cognitive impairment in DMD patients.

    “our finding showing downregulation of Dmd by miR-9-3p may provide new insights for the development of treatments for cognitive impairment in DMD patients.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    The miR-9-5p/3p gene is ancient in evolution and well conserved.

    “The miR-9-5p/3p gene is ancient in evolution and well conserved.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Wheeler et al., 2009 source

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

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

    Premisecitation

    The SAP97-myosin VI complex plays a critical role in trafficking of AMPARs to synapses.

    “Nash et al. (2010) showed that the SAP97-myosin VI complex plays a critical role in trafficking of AMPARs to synapses”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Nash et al., 2010

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

  • May combine two claims.

    Interpretationown data

    miR-9-3p is critically involved in hippocampal LTP and long-term memory.

    “Our results demonstrate that miR-9-3p is critically involved in hippocampal LTP and long-term memory.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    SAP97 is the only member of PSD-MAGUKs that binds directly to the C-terminus of GluA1 and is involved in AMPAR trafficking.

    “SAP97 is the only member of PSD-95-like membrane associated guanylate kinases (PSD-MAGUKs) that binds directly to the C-terminus of GluA1 and is involved in AMPA receptor (AMPAR) trafficking (Leonard et al., 1998;Cai et al., 2002).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Leonard et al., 1998 source
    • cites Cai et al., 2002 source

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

    Premisecitation

    The effects of SAP97 overexpression on synaptic transmission are influenced by isoforms of SAP97, compensation with other PSD-MAGUKs, or matureness of synapse.

    “The effects of SAP97 overexpression on synaptic transmission seem to be influenced by many factors, such as isoforms of SAP97 (Schlu ¨ter et al., 2006;Waites et al., 2009), compensation with other PSD-MAGUKs (Schlu ¨ter et al., 2006;Howard et al., 2010), or matureness of synapse (Howard et al., 2010).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Schlüter et al., 2006

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

    • cites Waites et al., 2009 source
    • cites Howard et al., 2010 source
18 of 18

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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.

  • Figure 1F names the wrong sponge in the LTD experiment

    Minor

    The Figure 1F caption says the LTD comparison used a miR-9-5p sponge, whereas the Results identify Figure 1F as the miR-9-3p sponge experiment.

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Authors
Sim, S.-E., Lim, C.-S., Kim, J.-I., Seo, D., Chun, H., Yu, N.-K., Lee, J., Kang, S. J., Ko, H.-G., Choi, J.-H., Kim, T., Jang, E.-H., Han, J., Bak, M. S., Park, J.-E., Jang, D.-J., Baek, D., Lee, Y.-S., & Kaang, B.-K.
Affiliations
  • Department of Biological Sciences (RU)
  • Institute for Basic Science (KR)
  • Chung-Ang University (KR)
  • Kyungpook National University (KR)
  • Seoul National University (KR)
ORCID
Venue
The Journal of Neuroscience
Publisher
Society for Neuroscience
Published
2016-08-17
Volume
36
Issue
33
Pages
8641-8652
ISSN
  • 0270-6474 (ISSN-L)
  • 1529-2401
DOI
10.1523/jneurosci.0630-16.2016
PMC
PMC6601897
OpenAlex
W2510924509
Semantic Scholar
f4ec49a6625afe4ee944887e877ccac25d3cd6bf
Cited by
116
References
67
Citations by year
2026: 4 · 2025: 12 · 2024: 18 · 2023: 11 · 2022: 16 · 2021: 10 · 2020: 15 · 2019: 15 · 2018: 8 · 2017: 7
FWCI
3.57
Type
journal article
Language
en
Open access
Locations
  • Journal of Neuroscience (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Seoul National University Open Repository (Seoul National University) (submittedVersion)
  • Scholarworks@UNIST (Ulsan National Institute of Science and Technology) (submittedVersion)
Funders
  • National Research Foundation
  • National Research Foundation of Korea (2012R1A3A1050385)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Neuroscience › Cognitive Neuroscience › Memory and Neural Mechanisms
Keywords
miR-9-3p, miR-9-5p, microRNA, synaptic plasticity, long-term potentiation, hippocampus, learning and memory, DMD, SAP97, post-transcriptional regulation, adult neurons, miRNA target genes
MeSH terms
Discs Large Homolog 1 Protein, Animals, Conditioning, Psychological, Exploratory Behavior, Fear, Hippocampus, Humans, Male, Membrane Proteins, Mice, Inbred C57BL, Neuronal Plasticity, Transduction, Genetic, Dystrophin, Synapsins, Maze Learning, Receptors, CXCR4, Recognition, Psychology, MicroRNAs, Green Fluorescent Proteins, Mice, Guanylate Kinases, HEK293 Cells

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