Cited by 374Neuroscience and Neuropharmacology ResearchMemory and Neural MechanismsNeural dynamics and brain function

Dynamic actin filaments are required for stable long-term potentiation (LTP) in area CA1 of the hippocampus

Open access
Krucker, Thomas, Siggins, George R., Halpain, Shelley·2000·Proceedings of the National Academy of Sciences

Abstract

The hypothesis that dynamic actin filaments participate in specific aspects of synaptic plasticity was investigated at the Schaffer-collateral-CA1 pyramidal cell synapse of mouse hippocampus. Low concentrations (0.01–1 μM) of compounds that inhibit actin filament assembly were bath applied to hippocampal slices during extracellular recording of field excitatory postsynaptic potentials. Cytochalasin D, cytochalasin B, and latrunculin A all impaired the maintenance of LTP induced by brief high-frequency stimulation. This effect on LTP maintenance was specific, because none of the compounds affected basal synaptic transmission, paired-pulse facilitation, LTP induction, or post-tetanic potentiation. The effect of cytochalasin B was reversible. The results are consistent with a model in which dynamic actin filaments play an essential role in the molecular mechanisms underlying the early maintenance phase of LTP, such as growth of new synaptic connections or conversion of silent synapses.

AI snapshot

Written from the paper’s full text.

TL;DR

Low concentrations of actin filament assembly inhibitors selectively impaired maintenance of LTP at Schaffer-collateral–CA1 synapses without affecting basal synaptic transmission, paired-pulse facilitation, LTP induction, or post-tetanic potentiation. The results suggest dynamic actin filaments are required during the early maintenance phase of LTP, possibly for structural remodeling or receptor trafficking.

Key findings

  • Cytochalasin D, cytochalasin B, and latrunculin A (0.01–1 μM) did not significantly alter basal pEPSPs, pharmacologically isolated NMDA-pEPSPs, presynaptic fiber volleys, or paired-pulse facilitation over 90 min.
  • In untreated slices, LTP was stable at 214 ± 17% of baseline at 100 min and 211 ± 20% at 130 min after tetanus. Pretreatment with actin assembly inhibitors reduced maintained LTP: cytochalasin D reached about 140% of baseline after 60 min, cytochalasin B about 148% at 130 min, and latrunculin A about 139% at 130 min.
  • When superfused after LTP induction, cytochalasin D reduced LTP to 165 ± 17% at 100 min and 121 ± 12% at 180 min; cytochalasin B to 182 ± 12% at 100 min and 146 ± 15% at 180 min; latrunculin A to 165 ± 3% at 100 min and 140 ± 12% at 180 min.
  • Cytochalasin B effects were at least partially reversible: after washout and a second tetanus, LTP was stable, and 0.01 μM cytochalasin B for 5 min reduced first LTP but allowed robust, nondecremental LTP of about 200% after the second tetanus.

Methods

  • Hippocampal slices (400 μm) from normal C57BL/6 male mice (32 ± 0.8 days old) were maintained in oxygenated ACSF at 32 ± 0.2°C.
  • Extracellular recordings of presynaptic fiber volleys and population EPSPs were made in stratum radiatum of CA1 with Schaffer-collateral stimulation; input–output curves and paired-pulse facilitation at 20, 50, 100, and 200 ms were tested.
  • LTP was induced by two tetani of 100 Hz, 20 sec apart, with responses followed up to 180 min; drugs were bath-applied before or immediately after LTP induction. pEPSP slope, amplitude, and area were analyzed, with Mann–Whitney U tests and P ≤ 0.05 considered significant.

Limitations

  • The paper states that the precise time window when dynamic actin filaments participate in LTP maintenance remains to be determined, and the precise nature and location of the essential actin filaments are not identified.
  • The authors state their studies do not rule out a role for dynamic actin in other regions of the neuron or in glial cells.
  • Evident from the text: experiments used only male C57BL/6 mice at one developmental age, in vitro slices, extracellular field recordings, and small group sizes (e.g., n = 3–7 per condition), which limits generalization and statistical power.

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

Integrity signals

Registry

Crossref · 10.1073/pnas.100139797

Cites

35 of 35 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 6 cited sources checked · 0 supported · 0 flagged

  • Findingown data
    § Abstract

    None of the compounds affected basal synaptic transmission.

    “none of the compounds affected basal synaptic transmission, paired-pulse facilitation, LTP induction, or post-tetanic potentiation.”
    Quote in the paperSubstantial claim
  • Findingown data
    § Results

    The presynaptic volley amplitude did not change during the entire recording time when AAIs were applied after LTP induction.

    “The presynaptic volley amplitude did not change during the entire recording time, suggesting that there were no modifications of the fiber input (data not shown).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The presynaptic volley amplitude is proportional to the number of presynaptic neurons recruited by stimulation.
  • Findingown data
    § Abstract

    None of the compounds affected post-tetanic potentiation.

    “none of the compounds affected basal synaptic transmission, paired-pulse facilitation, LTP induction, or post-tetanic potentiation.”
    Quote in the paperSubstantial claim
  • Findingown data
    § Abstract

    None of the compounds affected paired-pulse facilitation.

    “none of the compounds affected basal synaptic transmission, paired-pulse facilitation, LTP induction, or post-tetanic potentiation.”
    Quote in the paperSubstantial claim
  • Findingown data
    § Results

    A lower concentration of cytochalasin B (0.01 μM) superfused for 5 min also reduced the magnitude of LTP in a manner indistinguishable from the effect of higher concentrations.

    “Indeed, a lower concentration of cytochalasin B (0.01 μM, n = 5) superfused for 5 min also reduced the magnitude of LTP in a manner indistinguishable from the effect of higher concentrations.”
    Quote in the paperNot a substantial claim
  • May combine two claims. 13
  • Classified as probably not a substantial claim. 6
  • May combine two claims.

    Findingown data
    § Results

    After cytochalasin D superfusion shortly after LTP induction, LTP magnitude was 165 ± 17% of baseline at 100 min (significantly different from untreated slices, P < 0.015) and further declined to 121 ± 12% of baseline after 180 min.

    “At 100 min after LTP induction, LTP magnitude was 165 ± 17% of baseline (significantly different from untreated slices, P < 0.015) and further declined to 121 ± 12% of baseline after 180 min.”
    Quote in the paperSubstantial claim
  • May combine two claims.

    Findingown data
    § Results

    After 100 min of LTP induction, cytochalasin B and latrunculin A reduced average pEPSP slope to 182 ± 12% and 165 ± 3% of baseline, respectively; after 180 min, both reduced LTP magnitude significantly (P < 0.02) to 146 ± 15% and 140 ± 12% of baseline.

    “Like the cytochalasin D effect, after 100 min of LTP induction, cytochalasin B (0.1 μM, n = 3) and latrunculin A (0.1 μM, n = 5) reduced the average pEPSP slope to 182 ± 12% and 165 ± 3% of baseline, respectively. After 180 min, both drugs reduced LTP magnitude significantly (P < 0.02) to 146 ± 15% and 140 ± 12% of baseline, respectively.”
    Quote in the paperSubstantial claim
  • May combine two claims.

    Findingown data
    § Results

    Cytochalasin B pretreated slices showed reduced PTP (not significant), potentiated after 10 min to values similar to control, then declined steadily to about 148% of baseline 130 min after tetanus.

    “Slices incubated with cytochalasin B (0.1 μM, n = 5) also showed reduced PTP, although this was not significant. Responses were potentiated after 10 min to values similar to those in control slices but then declined steadily afterward and reached about 148% of baseline 130 min after tetanus.”
    Quote in the paperSubstantial claim
  • May combine two claims.

    Findingown data
    § Results

    Latrunculin A pretreated slices showed nonsignificantly reduced PTP, were potentiated for 10 min, and declined to about 139% of baseline 130 min after tetanus.

    “slices incubated with latrunculin A (0.1 μM, n = 5) showed nonsignificantly reduced PTP, were potentiated for 10 min, and declined to about 139% of baseline 130 min after tetanus.”
    Quote in the paperSubstantial claim
  • May combine two claims.

    Findingown data
    § Abstract

    Cytochalasin D, cytochalasin B, and latrunculin A all impaired the maintenance of LTP induced by brief high-frequency stimulation.

    “Cytochalasin D, cytochalasin B, and latrunculin A all impaired the maintenance of LTP induced by brief high-frequency stimulation.”
    Quote in the paperSubstantial claim
  • May combine two claims.

    Findingown data
    § Discussion

    The presynaptic fiber volley and PPF remain unchanged during AAI superfusion.

    “In addition, the presynaptic fiber volley and PPF remain unchanged.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The presynaptic volley amplitude is proportional to the number of presynaptic neurons recruited by stimulation.
  • May combine two claims.

    Findingown data
    § Results

    After the second set of tetani with 0.01 μM cytochalasin B, robust LTP was observed; it declined only slightly and reached a plateau of about 200% of control after 30 min.

    “However, we observed robust LTP after the second set of tetani. This enhancement declined only slightly and reached a plateau of about 200% of control after 30 min.”
    Quote in the paperSubstantial claim
  • May combine two claims.

    Interpretationown data
    § Discussion

    Dynamic actin filaments in the presynaptic compartment do not take part in basal synaptic transmission or in short-term plasticity.

    “our results also suggest that dynamic actin filaments in the presynaptic compartment do not take part in basal synaptic transmission or in short-term plasticity.”
    Quote in the paperSubstantial claim
  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductown data
    § Methods

    The final concentration of DMSO in the slice chamber never exceeded 0.1%, a concentration found in control experiments to have no detectable effects on the slices.

    “The final concentration of DMSO in the slice chamber never exceeded 0.1%, a concentration found in control experiments to have no detectable effects on the slices.”
    Quote in the paperNot a substantial claim
  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation
    § Results

    Latrunculins bind to G-actin in a 1:1 molar complex, thereby limiting the availability of actin monomers for polymerizing at either barbed or pointed ends of actin filaments.

    “Latrunculins bind to G-actin in a 1:1 molar complex, thereby limiting the availability of actin monomers for polymerizing at either barbed or pointed ends of actin filaments (4).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [4] Spector I, Shochet N R, Blasberger D, Kashman Y. Cell Motil Cytoskeleton. 1989;13:127–144. doi: 10.1002/cm.970130302. [DOI] [PubMed] [Google Scholar] source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisenone given
    § Results

    The actin inhibitors selectively disrupt filaments undergoing rapid length excursions, leaving intact the more stable and slowly-turning-over actin filaments.

    “These actin inhibitors selectively disrupt filaments undergoing rapid length excursions, leaving intact the more stable and slowly-turning-over actin filaments.”
    Quote in the paperSubstantial claim
  • Classified as probably not a substantial claim.

    Study design and conductnone given
    § Discussion

    The use of low but effective concentrations of actin inhibitors was designed to avoid nonspecific effects.

    “The use of low but effective concentrations of actin inhibitors was designed to avoid nonspecific effects.”
    Quote in the paperSubstantial claim
  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning
    § Discussion

    The studies do not rule out a role for dynamic actin in other regions of the neuron or in glial cells.

    “Indeed, whereas dynamic filaments in dendrites and spines are logical candidates, our studies do not rule out a role for dynamic actin in other regions of the neuron or in glial cells.”
    Quote in the paperNot a substantial claim
  • May combine two claims.

    Interpretationown data
    § Results

    The finding that LTP magnitude after second tetani with 0.01 μM cytochalasin B resembled normal levels and was nondecremental suggests that cytochalasin B effects are reversible.

    “Because this LTP magnitude resembled normal levels and was nondecremental, this finding suggests that cytochalasin B effects are reversible.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: Cytochalasin D exhibits poor washout in intact preparations, whereas cytochalasin B effects are more readily reversible.
  • May combine two claims.

    Premisecitation
    § Discussion

    Low concentrations of AAIs (0.01-1 μM) are similar to those shown previously to alter actin filament organization in cultured cells, prevent lamellipodial ruffling in fibroblasts, inhibit motility and retrograde flow in neuronal growth cones, and arrest dendritic spine motility in hippocampal neurons.

    “The concentrations of latrunculin A and cytochalasins used here (0.01–1 μM) are similar to those that have been shown previously to alter actin filament organization in cultured cells, prevent lamellipodial ruffling in fibroblasts, inhibit motility and retrograde flow in neuronal growth cones, and arrest dendritic spine motility in hippocampal neurons (3, 4, 7, 10).”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [3] Cooper J A. J Cell Biol. 1987;105:1473–1478. doi: 10.1083/jcb.105.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar] source

      Not checked in the free run.

    • cites [4] Spector I, Shochet N R, Blasberger D, Kashman Y. Cell Motil Cytoskeleton. 1989;13:127–144. doi: 10.1002/cm.970130302. [DOI] [PubMed] [Google Scholar] source

      Not checked in the free run.

    • cites [7] Forscher P, Smith S J. J Cell Biol. 1988;107:1505–1516. doi: 10.1083/jcb.107.4.1505. [DOI] [PMC free article] [PubMed] [Google Scholar] source

      Not checked in the free run.

    • cites [10] Fischer M, Kaech S, Knutti D, Matus A. Neuron. 1998;20:847–854. doi: 10.1016/s0896-6273(00)80467-5. [DOI] [PubMed] [Google Scholar] source

      Not checked in the free run.

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

    Premisecitation
    § Results

    At low concentrations, cytochalasins primarily cap the barbed ends of actin filaments.

    “at low concentrations, their primary effect in cells is to cap the barbed ends of actin filaments (3).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [3] Cooper J A. J Cell Biol. 1987;105:1473–1478. doi: 10.1083/jcb.105.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar] source

      Not checked in the free run.

  • May combine two claims.

    Premisereasoning
    § Discussion

    Under the conditions used here, the application of actin perturbing agents is not expected to disrupt stable actin filaments or induce collapse of dendritic spines.

    “Under the conditions used here, the application of actin perturbing agents is not expected to disrupt stable actin filaments or induce collapse of dendritic spines.”
    Quote in the paperSubstantial claim
17 of 17
  • The paper relies on this without support.

    Cytochalasin D exhibits poor washout in intact preparations, whereas cytochalasin B effects are more readily reversible.

    “It has been reported that cytochalasin D exhibits poor washout in intact preparations, but that effects of cytochalasin B are more readily reversible.”
  • The paper relies on this without support.

    The presynaptic volley amplitude is proportional to the number of presynaptic neurons recruited by stimulation.

    “the amplitude of the presynaptic volley, which is proportional to the number of presynaptic neurons recruited by stimulation”
2 of 2

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.

  • Input-output curve times differ between Methods and Results

    Major

    The Methods say additional input-output curves were generated 20 and 40 minutes after drug superfusion began, whereas the Results place them at 40 and 80 minutes.

  • Reversibility conclusion extends beyond the reported drug effects

    Minor

    The Discussion says washout showed that inhibition by actin assembly inhibitors was reversible, but the Results distinguish cytochalasin B as at least partially reversible in contrast to cytochalasin D and latrunculin A.

2 of 2

Cites

Flagged first, then by citations

1 of the 36 references couldn’t be matched to a known paper.

Research works printed but not linked: 1

  1. H Wigstrom, B Gustafsson J Physiol 81, 228–236 (1986).

Cited by

Sorted by citations

Authors
Krucker, T., Siggins, G. R., & Halpain, S.
Affiliations
Scripps Research Institute (US)
Venue
Proceedings of the National Academy of Sciences
Published
2000-05-23
Volume
97
Issue
12
Pages
6856-6861
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
OpenAlex
W2041963785
Cited by
374
References
36
Citations by year
2026: 6 · 2025: 4 · 2024: 7 · 2023: 8 · 2022: 1 · 2021: 11 · 2020: 7 · 2019: 5 · 2018: 10 · 2017: 8 · 2016: 10 · 2015: 10 · 2014: 17 · 2013: 23 · 2012: 23
FWCI
6.96
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institute of Mental Health (R01MH050861, MH47680, P50MH047680)
  • National Institute of Neurological Disorders and Stroke (R01NS037311, NS37311)
Topics
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Neuroscience › Cognitive Neuroscience › Memory and Neural Mechanisms
  • Life Sciences › Neuroscience › Cognitive Neuroscience › Neural dynamics and brain function
Keywords
F-actin, long-term potentiation, hippocampus, CA1 pyramidal neurons, Schaffer collateral synapses, synaptic plasticity, cytochalasin B, cytochalasin D, latrunculin A, actin filament assembly, silent synapses, field excitatory postsynaptic potentials
MeSH terms
Actins, Animals, Cytochalasin B, Hippocampus, Male, Mice, Inbred C57BL, Synaptic Transmission, Neuronal Plasticity, Long-Term Potentiation, Mice

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