Dynamic actin filaments are required for stable long-term potentiation (LTP) in area CA1 of the hippocampus
Open accessAbstract
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.
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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.
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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
- Findingown data§ Abstract
None of the compounds affected LTP induction.
“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
Control (untreated) slices exhibited stable LTP of 214 ± 17% of baseline at 100 min and 211 ± 20% at 130 min after tetanus.
“Control (untreated) slices exhibited stable LTP of 214 ± 17% of baseline at 100 min and 211 ± 20% at 130 min after tetanus.”
Quote in the paperSubstantial claim - Findingown data§ Results
All LTP magnitudes at 100 and 130 min were significantly reduced relative to control (P < 0.012) in slices pretreated with AAIs.
“However, all LTP magnitudes at 100 and 130 min were significantly reduced relative to control (P < 0.012).”
Quote in the paperSubstantial claim - Findingnone given§ Results
Pretreatment with AAIs impairs LTP.
“Pretreatment with Actin Assembly Inhibitors Impairs LTP.”
Quote in the paperSubstantial claim - Findingown data§ Results
Cytochalasin D pretreated slices showed reduced PTP, potentiated to about 200% of baseline after 10 min, then declined steadily to reach a plateau of about 140% of baseline after 60 min of LTP induction.
“slices incubated with cytochalasin D (1 μM, n = 4; 0.1 μM, n = 3; data pooled) showed reduced PTP but were subsequently potentiated to about 200% of baseline after 10 min, then declined steadily to reach a plateau of about 140% of baseline after 60 min of LTP induction.”
Quote in the paperSubstantial claim - Findingown data§ Results
After a second LTP induction protocol following cytochalasin D washout, the pEPSP was potentiated for about 10 min but steadily declined to reach 113 ± 10% and 130 ± 21% of control.
“After the second LTP induction protocol, the pEPSP was potentiated for about 10 min but steadily declined to reach 113 ± 10% and 130 ± 21%, respectively, of control.”
Quote in the paperSubstantial claim - Findingown data§ Results
When AAIs were superfused shortly after LTP induction, PTP remained unchanged.
“In contrast to slices pretreated with AAIs, where PTP was noticeably affected and could have influenced LTP expression, PTP remained unchanged when AAIs were superfused shortly after LTP induction.”
Quote in the paperSubstantial claim - Findingown data§ Abstract
The effect of cytochalasin B on LTP was reversible.
“The effect of cytochalasin B was 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.
- Findingown data§ Discussion
Continuous superfusion of AAIs for over 90 min affects neither pEPSPs nor pharmacologically isolated NMDA-pEPSPs over a series of stimulation intensities.
“We have shown that continuous superfusion of AAIs for over 90 min affects neither pEPSPs nor pharmacologically isolated NMDA–pEPSPs over a series of stimulation intensities.”
Quote in the paperSubstantial claim - Findingown data§ Results
Cytochalasin D superfused for 5 or 10 min significantly reduced LTP magnitude after 80 min, similar to continuous superfusion for 60-80 min.
“Both concentrations and application times significantly reduced LTP magnitude after 80 min, similar to when they were superfused continuously for 60–80 min (compare Fig. 4A to Fig. 3C).”
Quote in the paperSubstantial claim - Interpretationown data§ Discussion
Dynamic actin filaments do not influence postsynaptic cellular mechanisms required for synaptic transmission during basal synaptic activity.
“We therefore conclude that dynamic actin filaments do not influence postsynaptic cellular mechanisms required for synaptic transmission during basal synaptic activity.”
Quote in the paperSubstantial claim - Interpretationown data§ Results
LTP maintenance is selectively blocked by AAIs.
“LTP Maintenance Is Selectively Blocked by Actin Assembly Inhibitors.”
Quote in the paperSubstantial claim - Interpretationown data§ Abstract
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.
“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.”
Quote in the paperSubstantial claim - Interpretationown data§ Discussion
Dynamic actin filaments are required shortly after the initial induction phase of LTP.
“The fact that a transient (5–10 min) exposure of slices to AAIs immediately following the tetanus was sufficient to impair the maintenance of LTP, similar to that seen with prolonged exposures, suggests that dynamic actin filaments are required shortly after the initial induction phase of LTP.”
Quote in the paperSubstantial claim - Interpretationown data§ Discussion
A resumption of actin filament turnover reestablishes conditions permissive for the maintenance of LTP.
“We therefore conclude that a resumption of actin filament turnover reestablishes conditions permissive for the maintenance of LTP.”
Quote in the paperSubstantial claim
- May combine two claims. 13
- Classified as probably not a substantial claim. 6
May combine two claims.
Findingown data§ ResultsAfter 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 claimMay combine two claims.
Findingown data§ ResultsAfter 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 claimMay combine two claims.
Findingown data§ ResultsCytochalasin 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 claimMay combine two claims.
Findingown data§ ResultsLatrunculin 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 claimMay combine two claims.
Findingown data§ AbstractCytochalasin 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 claimMay combine two claims.
Findingown data§ DiscussionThe 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§ ResultsAfter 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 claimMay combine two claims.
Interpretationown data§ DiscussionDynamic 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 claimProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Study design and conductown data§ MethodsThe 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 claimProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Premisecitation§ ResultsLatrunculins 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§ ResultsThe 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 claimClassified as probably not a substantial claim.
Study design and conductnone given§ DiscussionThe 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 claimProbably not a substantial claim: may not pass S3 (load-bearing).
Interpretationreasoning§ DiscussionThe 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 claimMay combine two claims.
Interpretationown data§ ResultsThe 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§ DiscussionLow 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§ ResultsAt 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§ DiscussionUnder 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
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”
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?
Actin polymerization is an essential step of LTP-stabilizing processes.
“Actin polymerization is an essential step of this kind (9, 15–17)”
From Synaptic evidence for the efficacy of spaced learning (2012) · cites this paper as [17]
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
MajorThe 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.
“To assess drug effects on synaptic efficacy, we generated additional I-O curves at 20 and 40 min after the start of drug superfusion with the same stimulation intensities as those under control conditions.”
“After 40 and 80 min of cytochalasin D superfusion, additional I-O curves performed at the same stimulation intensities as under control conditions also showed no detectable changes (Fig. 2A).”
Reversibility conclusion extends beyond the reported drug effects
MinorThe 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.
“In addition, drug wash-out experiments show that the inhibition of LTP by AAIs is reversible.”
“Similarly, we observed that, in contrast to the effects of cytochalasin D and latrunculin A, the effects of cytochalasin B on LTP appeared to be at least partially reversible (Fig. 4C).”
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- Authors
- Krucker, T., Siggins, G. R., & Halpain, S.
- Affiliations
- Scripps Research Institute (US)
- ORCID
- Halpain, Shelley
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2000-05-23
- Volume
- 97
- Issue
- 12
- Pages
- 6856-6861
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC18765
- OpenAlex
- W2041963785
- Semantic Scholar
- 879b8456d922579733d332c78244f89936f74a66
- 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
- green
- www.ncbi.nlm.nih.gov
- 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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Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-10.
Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-10.
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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