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Cited by 38Cellular transport and secretionNerve injury and regenerationNeuroscience and Neuropharmacology Research

The DOI matches its Crossref record; 69 of the 70 references it cites were found in a registry or an index; 8 of the 24 sources its claims cite were checked for support.

The Rab5–Rab11 Endosomal Pathway is Required for BDNF-Induced CREB Transcriptional Regulation in Hippocampal Neurons

Open access
González-Gutiérrez, Andrés, Lazo, Oscar M., Bronfman, Francisca C.·2020·The Journal of Neuroscience
DOI 10.1523/jneurosci.2063-19.2020PMC PMC7574648

Published version of a preprint

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Abstract

Brain-derived neurotrophic factor (BDNF) is a key regulator of the morphology and connectivity of central neurons. We have previously shown that BDNF/TrkB signaling regulates the activity and mobility of the GTPases Rab5 and Rab11, which in turn determine the postendocytic sorting of signaling TrkB receptors. Moreover, decreased Rab5 or Rab11 activity inhibits BDNF-induced dendritic branching. Whether Rab5 or Rab11 activity is important for local events only or for regulating nuclear signaling and gene expression is unknown. Here, we investigated, in rat hippocampal neuronal cultures derived from embryos of unknown sex, whether BDNF-induced signaling cascades are altered when early and recycling endosomes are disrupted by the expression of dominant-negative mutants of Rab5 and Rab11. The activity of both Rab5 and Rab11 was required for sustained activity of Erk1/2 and nuclear CREB phosphorylation, and increased transcription of a BDNF-dependent program of gene expression containing CRE binding sites, which includes activity-regulated genes such as Arc , Dusp1 , c-fos , Egr1 , and Egr2 , and growth and survival genes such as Atf3 and Gem . Based on our results, we propose that early and recycling endosomes provide a platform for the integration of neurotrophic signaling from the plasma membrane to the nucleus in neurons, and that this mechanism is likely to regulate neuronal plasticity and survival. SIGNIFICANCE STATEMENT BDNF is a neurotrophic factor that regulates plastic changes in the brain, including dendritic growth. The cellular and molecular mechanisms underlying this process are not completely understood. Our results uncover the cellular requirements that central neurons possess to integrate the plasma membrane into nuclear signaling in neurons. Our results indicate that the endosomal pathway is required for the signaling cascade initiated by BDNF and its receptors at the plasma membrane to modulate BDNF-dependent gene expression and neuronal dendritic growth mediated by the CREB transcription factor. CREB is a key transcription factor regulating circuit development and learning and memory.

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

BDNF/TrkB signaling in hippocampal neurons requires Rab5 and Rab11 endosomal activity for sustained Erk1/2 activation, nuclear CREB phosphorylation, and CRE-dependent gene expression. Disrupting these GTPases blocks BDNF-induced CREB activation and dendritic branching, linking endosomal trafficking to plasma-membrane-to-nucleus signaling.

Key findings

  • Five minutes of BDNF stimulation activated TrkB, Akt, and Erk1/2 similarly in control EGFP, Rab5DN, and Rab11DN neurons; after 120 minutes, pErk1/2 remained elevated in EGFP neurons but returned to basal levels in Rab5DN- and Rab11DN-expressing neurons.
  • Rab5DN and Rab11DN blocked BDNF-induced CREB phosphorylation at S133 by western blot and nuclear immunofluorescence at 15–60 minutes. PD98059 blocked nuclear pCREB, K252a blocked it, but LY294002 did not affect pCREB despite blocking Akt phosphorylation at S473.
  • Expression of non-phosphorylatable CREB S133A abolished BDNF-induced increases in dendritic complexity by Sholl analysis and branching points.
  • BDNF increased Arc expression 5.4-fold in EGFP neurons versus 3.3-fold in Rab11DN neurons. In a CRE/SRE/CaRE qPCR array, 76 of 84 genes were reliably analyzed, 19 were BDNF-regulated, and 9 were significantly reduced by Rab11DN: Dusp1, Gem, Nf1, Atf3, Scg2, Egr1, Fos, Egr2, and Calcrl.

Methods

  • Primary hippocampal neurons from E18 rat embryos were cultured and transduced at 7 DIV with control EGFP, Rab5DN, or Rab11DN adenovirus, or transfected with CREB S133A; experiments were performed 48 hours later.
  • BDNF-stimulated neurons were analyzed by western blotting for phospho-TrkB, Akt, Erk1/2, and CREB; by confocal immunofluorescence for pErk1/2 and pCREB in nuclei, somas, and dendrites; by qPCR for Arc and an 84-gene CRE/SRE/CaRE array; and by Sholl analysis for dendritic arborization.
  • Statistical analysis used Student’s t-test, one-way ANOVA with multiple-comparison post-tests, or two-way repeated-measures ANOVA for Sholl curves, with significance reported in figure legends.

Limitations

  • The paper states that the precise mechanism by which signaling endosomes facilitate pErk1/2 nuclear translocation needs further investigation, and it proposes future studies on the BDNF–Rab5/Rab11–Erk1/2–CREB–c-fos–Egr1 pathway in long-term memory and dendritic morphology maintenance.
  • Evident from the text: all experiments are in cultured rat hippocampal neurons, not in vivo; the main genetic tools are dominant-negative Rab5/Rab11 mutants; gene-expression analysis after Rab11DN focuses on Rab11 rather than Rab5; and the qPCR array yielded reliable data for 76 of 84 genes.

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

Integrity signals

Registry

Crossref · 10.1523/jneurosci.2063-19.2020

Cites

69 of 69 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Verification

Claims the paper makes

8 of 24 cited sources checked · 5 supported · 0 flagged · accuracy not yet measured

  • Interpretationown data

    Endocytic pathways regulate the propagation of BDNF signaling to the nucleus in addition to the local events reported previously.

    “Altogether, our results suggest that endocytic pathways regulate the propagation of BDNF signaling to the nucleus in addition to the local events reported previously (Huang et al., 2013;Lazo et al., 2013;Moya-Alvarado et al., 2018).”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites Huang et al., 2013 source
    • cites Lazo et al., 2013 source
    • cites Moya-Alvarado et al., 2018 source
  • Findingown data

    In control cells, Akt activity returned to basal levels by 120 minutes of BDNF stimulation.

    “Akt activity showed a normal return to basal levels by 120 minutes of BDNF stimulation in control cells.”
    Quote in the paperNot a substantial claim
  • Findingown data

    Neurons expressing Rab5DN or Rab11DN and stimulated with BDNF for 60 minutes showed a decreased response to BDNF.

    “Consistent with the results of Fig. 1, neurons expressing Rab5DN or Rab11DN and stimulated with BDNF for 60 minutes showed a decreased response to BDNF (Fig. 2D and E).”
    Quote in the paperSubstantial claim
  • Findingown data

    PD98059 reduces pErk1/2 staining in somas and nuclei.

    “The ability of PD98059 to reduce pErk1/2 staining was evident in somas and nuclei, and the quantifications are shown in (B-C).”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: PD98059 specifically inhibits MEK1/2 without off-target effects on other kinases.
  • Findingown data

    The subset of BDNF-regulated genes that depends on endocytic trafficking through the recycling pathway encompasses diverse functional groups involved in metabolism, neurotransmission and regulation of transcription.

    “The subset of BDNF-regulated genes that depends on endocytic trafficking through the recycling pathway encompasses diverse functional groups involved in metabolism, neurotransmission and regulation of transcription, among others.”
    Quote in the paperNot a substantial claim
  • It may state two separate claims as one. 21
  • Probably not a substantial claim. 11
  • The quote only partly states this claim. 3

  • It may state two separate claims as one.

    Premisecitation

    Rab5 activity is required for maintenance of basal dendritic complexity and for the increase in dendritic complexity induced by BDNF in hippocampal neurons.

    “We have previously shown that Rab5 activity is required for both the maintenance of basal dendritic complexity and for the increase in dendritic complexity induced by BDNF in hippocampal neurons.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites Lazo et al., 2013 source

      The Results section, “BDNF-induced dendritic branching requires Rab11” (page unavailable), reports that inhibiting Rab11 abolished BDNF-dependent dendritic growth in hippocampal neurons. A search for Rab5 and basal dendritic complexity found no passage establishing the attributed requirement for Rab5; the Rab11 finding does not rule it out. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites Moya-Alvarado et al., 2018 source

  • It may state two separate claims as one.

    Premisecitation

    Rab11DN-expressing neurons did not show a decrease in basal dendritic complexity at 7 DIV and are specifically required for BDNF-induced dendritic branching.

    “Rab11DN-expressing neurons, however, did not show a decrease in basal dendritic complexity at 7 DIV and are thus specifically required for BDNF-induced dendritic branching (Lazo et al., 2013;Moya-Alvarado et al., 2018).”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites Lazo et al., 2013 source

      The Results section ('BDNF-induced dendritic branching requires Rab11'; page unavailable) reports that Rab11DN abolished BDNF-dependent dendritic growth in hippocampal neurons treated at 7 DIV. It does not state that Rab11DN-expressing neurons had unchanged basal dendritic complexity. The Methods section says arborization was assessed 48 hours after treatment, rather than at 7 DIV. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites Moya-Alvarado et al., 2018 source

  • Probably not a substantial claim. It may state two separate claims as one.

    Premisecitation

    All these genes, including Bdnf itself, are immediate early genes whose transcription is upregulated by activation of CREB in the context of increased neuronal activity.

    “All these genes, including Bdnf itself, are immediate early genes whose transcription is upregulated by activation of CREB in the context of increased neuronal activity (Flavell and Greenberg, 2008;Yap and Greenberg, 2018).”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites Flavell and Greenberg, 2008 source

      In the Introduction (page unavailable), the paper identifies bdnf as an activity-regulated CREB target and reports that CREB binding contributes to activation of its promoter IV after neuronal calcium influx. It does not establish that all the genes referred to are immediate early genes. It distinguishes bdnf’s slower transcriptional response from the rapid response characteristic of immediate early genes. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites Yap and Greenberg, 2018 source

  • It may state two separate claims as one.

    Findingown data

    BDNF increased Arc in EGFP-expressing neurons by 5.4-fold, while Rab11DN-expressing neurons displayed an attenuated response (3.3-fold).

    “BDNF increased Arc in EGFP-expressing neurons by 5.4-fold (Fig. 5A), while Rab11DN-expressing neurons displayed an attenuated response (3.3-fold), suggesting that Rab11 significantly mediates transcriptional response to neurotrophic signaling.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    EGFP-expressing neurons responded to BDNF stimulation with a significant increase in pCREB levels, while expression of DN mutants of Rab5 and Rab11 completely blocked this effect.

    “EGFP-expressing neurons responded to BDNF stimulation with a significant increase in pCREB levels, while expression of DN mutants of Rab5 and Rab11 completely blocked this effect (Fig. 3A and B).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingcitation

    BDNF stimulation induces a strong increase in nuclear pCREB, which is blocked by Trk inhibitor K252a.

    “BDNF stimulation induces a strong increase in nuclear pCREB, which is blocked by the Trk inhibitor K252a (Tapley et al., 1992).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Tapley et al., 1992

  • It may state two separate claims as one.

    Findingown data

    After 120 minutes of BDNF, phosphorylated Erk1/2 levels were still significantly increased in EGFP-expressing neurons, while cells expressing Rab5DN and Rab11DN showed levels that decreased to basal levels.

    “However, after 120 minutes, the levels of phosphorylated Erk1/2 were still significantly increased in EGFP-expressing neurons, while cells expressing Rab5DN and Rab11DN showed levels that decreased to the basal levels (Fig. 1B).”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Findingcitation

    Atf3 was upregulated by BDNF and downregulated by reducing Rab11 activity.

    “The Atf3 gene product is a TF involved in neuronal survival, and it was also upregulated by BDNF and downregulated by reducing Rab11 activity (Wu et al., 2013).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    BDNF-induced increase in dendritic complexity (Sholl's analysis) and increase in number of branches were completely abolished by CREB S133A.

    “However, the BDNFinduced increase in complexity, as measured by using Sholl's analysis (Fig. 4E), and, more specifically, the increase in the number of branches (Fig. 3F) were completely abolished by the presence of the CREB S133A mutant, suggesting that CREB is needed for BDNF-dependent dendritic branching downstream of TrkB and Erk1/2.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The CREB S133A mutant is non-phosphorylatable and acts to block CREB-mediated transcription.

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Findingown data

    PD98059 inhibits BDNF-induced phosphorylation of Erk1/2.

    “The ability of PD98059 to inhibit BDNF-induced phosphorylation of Erk1/2 was confirmed by stimulating hippocampal neurons with BDNF for 15 minutes and performing double immunofluorescence for MAP2 and pErk1/2 (A).”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: PD98059 specifically inhibits MEK1/2 without off-target effects on other kinases.

  • It may state two separate claims as one.

    Findingown data

    Rab11DN significantly decreased Erk1/2 activity in the soma and completely eliminated it in the nucleus; effect in dendrites was unclear.

    “Although the effect of Rab11DN in dendrites was unclear, the activity of Erk1/2 in the soma was significantly decreased and completely eliminated in the nucleus by this mutant (Fig. 2D and E).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Of the 64 genes analyzed containing CRE binding sites, Egr1 and Egr2 were the most highly upregulated by BDNF and also downregulated by reduced Rab11 activity.

    “Of note, of the 64 genes analyzed containing CRE binding sites, Egr1 and Egr2 were the most highly upregulated by BDNF and also downregulated by reduced Rab11 activity (Fig. 5B and C).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    LY294002 blocked Akt phosphorylation at serine 473 without affecting BDNF-dependent Trk phosphorylation.

    “We confirmed that although BDNFdependent Trk phosphorylation was independent of the presence of the inhibitor, Akt phosphorylation at serine 473 was completely blocked.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. The quote only partly states this claim.

    Findingown data

    76 genes were reliably analyzed from a qPCR array for 84 genes regulated by CRE, SRE and CaRE response elements.

    “We reliably analyzed 76 genes (Supplementary Table S1);”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    In most cases, hippocampal neurons expressing Rab11DN showed an attenuated response to BDNF, and 9 genes were significantly reduced (Dusp1, Gem, Nf1, Atf3, Scg2, Egr1, Fos, Egr2 and Calcrl).

    “In most cases, hippocampal neurons expressing Rab11DN showed an attenuated response to BDNF, and 9 of these genes were significantly reduced (Dusp1, Gem, Nf1, Atf3, Scg2, Egr1, Fos, Egr2 and Calcrl) (Fig. 5C).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Control EGFP expressing neurons showed a strong increase in CREB activation after 15-60 minutes of BDNF stimulation; Rab5DN and Rab11DN significantly blocked BDNF-induced activation of CREB in the nucleus.

    “While control EGFP expressing neurons showed a strong increase in CREB activation after 15-60 minutes of stimulation (Fig 3C-F), the expression of Rab5DN and Rab11DN significantly blocked BDNF-induced activation of CREB in the nucleus (Fig 3C-F), consistent with the effects on nuclear pErk1/2.”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    In a PCR array of 84 CRE/SRE/CaRE-regulated genes, 19 genes show significant expression differences after BDNF stimulation.

    “Reliable data obtained from 76 of the genes in 5 independent experiments were categorized by their annotated functions and plotted as a fold-change in BDNF-stimulated neurons compared to BSA-treated neurons, resulting in significant differences in 19 of these genes (B).”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    Overexpression of a constitutively active mutant of CREB led to increased expression of BDNF and BDNF-dependent long-term memory in the hippocampus.

    “Indeed, the overexpression of a constitutively active mutant of CREB led to increased expression of BDNF and BDNF-dependent long-term memory in the hippocampus (Barco et al., 2005).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Barco et al., 2005 source

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

  • It may state two separate claims as one.

    Interpretationreasoning

    BDNF/TrkB signaling activates nuclear CREB through the Erk1/2 pathway, requiring an increase in the activities of Rab5 and Rab11 to regulate gene expression.

    “Taken together, our results suggest that BDNF/TrkB signaling activates nuclear CREB through the Erk1/2 pathway, requiring an increase in the activities of Rab5 and Rab11 to regulate gene expression.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Expression of Rab5DN and Rab11DN mutants specifically inhibits Rab5 and Rab11 activity without affecting other cellular processes.
    • Rests on an assumption: Phosphorylation of CREB at S133 is a valid readout of CREB transcriptional activation.

  • It may state two separate claims as one.

    Interpretationreasoning

    The Rab11 recycling endosomes constitute a platform for stabilization and integration of signaling receptors, avoiding degradation and allowing intracellular signaling and neuronal function.

    “Thus, the Rab11 recycling endosomes constitute a platform for stabilization and integration of signaling receptors, avoiding degradation and allowing intracellular signaling and neuronal function.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Sustained activation of Erk1/2 is crucial for structural plasticity in hippocampal neurons.

    “Consistently, sustained activation of Erk1/2 is crucial for structural plasticity in hippocampal neurons (Wu et al., 2001b).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Wu et al., 2001b

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

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    These genes include early-response genes, such as Egr1, Egr2, Fos and Arc, that are important in brain plasticity and memory formation.

    “These genes include early-response genes, such as Egr1, Egr2, Fos and Arc, that are important in brain plasticity and memory formation (Heroux et al., 2018;Minatohara et al., 2015).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Heroux et al., 2018

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

    • cites Minatohara et al., 2015

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

  • It may state two separate claims as one.

    Interpretationown data

    Downregulation of CREB nuclear activation was not due to a reduction in the basal levels of the receptor or signaling molecules since 48 hours of expression of the Rab5DN and Rab11DN mutants did not preclude the activation of the receptor or the engagement of the PI3K-Akt and Erk1/2 signaling pathways after 5 minutes of BDNF stimulation.

    “Downregulation of CREB nuclear activation was not due to a reduction in the basal levels of the receptor or signaling molecules since 48 hours of expression of the Rab5DN and Rab11DN mutants did not preclude the activation of the receptor or the engagement of the PI3K-Akt and Erk1/2 signaling pathways after 5 minutes of BDNF stimulation.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    The Atf3 gene product is a TF involved in neuronal survival.

    “The Atf3 gene product is a TF involved in neuronal survival, and it was also upregulated by BDNF and downregulated by reducing Rab11 activity (Wu et al., 2013).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Wu et al., 2013

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

  • It may state two separate claims as one.

    Interpretationreasoning

    The BDNF-Rab5/Rab11-Erk1/2-CREB pathway also triggers genes that downregulate neurotrophic responses, providing a mechanism for homeostatic regulation.

    “Thus, our results suggest that the BDNF-Rab5/Rab11-Erk1/2-CREB pathway also triggers genes that downregulate neurotrophic responses, providing a mechanism for homeostatic regulation.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductnone given

    Adenoviral transduction of hippocampal neurons achieved infection efficiency over 80%.

    “transduced them with adenoviruses to achieve infection efficiency over 80%”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Arc in neurons is associated with endocytosis of AMPA receptors that is required for synaptic depression.

    “However, one of the role associated with Arc in neurons is the endocytosis of AMPA receptors that is required for synaptic depression (Barylko et al., 2018).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Barylko et al., 2018

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

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. The quote only partly states this claim.

    Study design and conductown data

    Reliable data were obtained from 76 of the 84 genes in the PCR array.

    “Reliable data obtained from 76 of the genes in 5 independent experiments”
    Quote in the paperNot a substantial claim
28 of 28

  • The paper relies on this without support.

    Expression of Rab5DN and Rab11DN mutants specifically inhibits Rab5 and Rab11 activity without affecting other cellular processes.

    “We used dominant negative (DN) mutants of Rab5 and Rab11 to investigate whether early and recycling endosomes play a role in sustained BDNF signaling.”

  • The paper relies on this without support.

    Phosphorylation of CREB at S133 is a valid readout of CREB transcriptional activation.

    “the levels of phosphorylated CREB at the S133 residue were evaluated by western blotting”

  • The paper relies on this without support.

    The CREB S133A mutant is non-phosphorylatable and acts to block CREB-mediated transcription.

    “we expressed a non-phosphorylatable CREB mutant (CREB S133A) in 7 DIV hippocampal neurons and stimulated them with BDNF”

  • The paper relies on this without support.

    Dominant negative mutants of Rab5 and Rab11 specifically inhibit Rab5 and Rab11 activity without affecting other pathways.

    “Hippocampal neurons expressing control EGFP or dominant negative (DN) mutants of Rab5 and Rab11 were stimulated with BDNF for 0, 5 or 120 minutes”

  • The paper relies on this without support.

    EGFP expression alone does not affect BDNF signaling, CREB phosphorylation, or dendritic branching.

    “Hippocampal neurons expressing control EGFP or dominant negative (DN) mutants of Rab5 and Rab11 were stimulated with BDNF for 0, 5 or 120 minutes”

  • The paper relies on this without support.

    The non-phosphorylatable CREB mutant (S133A) specifically prevents CREB phosphorylation at S133 without other effects.

    “we compared BDNF-induced dendritic branching in neurons expressing either EGFP or a non-phosphorylatable mutant of CREB (S133A).”

  • The paper relies on this without support.

    PD98059 specifically inhibits MEK1/2 without off-target effects on other kinases.

    “the MEK1/2 inhibitor PD98059”

  • The paper relies on this without support.

    LY294002 specifically inhibits PI3K without off-target effects.

    “the PI3K inhibitor LY294002”

  • The paper relies on this without support.

    K252a specifically inhibits Trk kinase activity without off-target effects.

    “the Trk kinase activity inhibitor K252a”

  • The paper relies on this without support.

    The housekeeping genes used for normalization are stably expressed across conditions.

    “The expression of β-actin, Tbp and Pgk-1 was used to normalize the data between independent experiments.”

  • The paper relies on this without support.

    The antibodies used are specific for their target proteins.

    “After electrophoretic transfer, nitrocellulose membranes were incubated with the following antibodies:”
11 of 11

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.

  • Branch counts attributed to a CREB-phosphorylation panel

    Minor

    The Results cite Figure 3F for the number of dendritic branches, but the Figure 3 legend identifies panels D-F as measurements of nuclear phosphorylated CREB. The Figure 4 legend instead assigns branching-point counts to panel I.

  • Figure 3 CREB time points disagree

    Minor

    The Results say nuclear CREB was measured at 30 minutes, but the Figure 3 legend lists only 0, 15 and 60 minutes for that experiment.

  • Sholl analysis assigned conflicting Figure 4 panels

    Minor

    The Results cite Figure 4E for Sholl analysis, whereas the Figure 4 legend assigns Sholl analysis to panel H and describes panel E as a pCREB measurement.

3 of 3

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Metadata

Authors
González-Gutiérrez, A., Lazo, O. M., & Bronfman, F. C.
Affiliations
  • UK Dementia Research Institute (GB)
  • Universidad Andrés Bello (CL)
Corresponding author
Bronfman, Francisca C.
ORCID
Venue
The Journal of Neuroscience
Publisher
Society for Neuroscience
Published
2020-09-14
Volume
40
Issue
42
Pages
8042-8054
ISSN
  • 0270-6474 (ISSN-L)
  • 1529-2401
DOI
10.1523/jneurosci.2063-19.2020
PMC
PMC7574648
OpenAlex
W3086481732
Cited by
38
References
70
Citations by year
2026: 6 · 2025: 10 · 2024: 2 · 2023: 9 · 2022: 10 · 2021: 1
FWCI
1.78
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Neuroscience (publishedVersion, cc-by-nc-sa)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nd)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • UCL Discovery (University College London) (submittedVersion)
  • Universidad de Chile (submittedVersion, cc-by-nc-nd)
Funders
  • Agencia Nacional de Investigacion y Desarrollo Chile (1171137)
  • Basal Center of Excellence in Science and Technology CONICYT (AFB170005)
  • Millenum Nucleus (P07/011-F)
  • MINEDUC | Comisión Nacional de Investigación Científica y Tecnológica (PhD Fellowshio)
  • MINEDUC | CONICYT | Fondo de Fomento al Desarrollo Científico y Tecnológico (Becas Chile)
  • Royal Society (Postdoc Fellowship)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Nerve injury and regeneration
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
Keywords
brain-derived neurotrophic factor, TrkB receptor, Rab5, Rab11, endocytic pathway, early endosomes, recycling endosomes, CREB phosphorylation, ERK signaling, hippocampal neurons, dendritic branching, transcriptional regulation, dominant-negative mutant, neuroplasticity, neuronal survival
MeSH terms
rab11 GTP-Binding Proteins, Animals, Dendrites, Gene Expression Regulation, Hippocampus, Neurons, Phosphorylation, Signal Transduction, Cyclic AMP Response Element-Binding Protein, Brain-Derived Neurotrophic Factor, rab GTP-Binding Proteins, rab5 GTP-Binding Proteins, MAP Kinase Signaling System, Rats, Primary Cell Culture

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