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Cited by 181Nerve injury and regenerationStroke Rehabilitation and RecoveryNeurogenesis and neuroplasticity mechanisms

GDF10 is a signal for axonal sprouting and functional recovery after stroke

Open access
Li, Songlin, Nie, Esther H, Yin, Yuqin, Benowitz, Larry I, Tung, Spencer, Vinters, Harry V, Bahjat, F Rena, Stenzel-Poore, Mary P +3 more·2015·Nature Neuroscience
DOI 10.1038/nn.4146PMC PMC4790086
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Abstract

Stroke produces a limited process of neural repair. Axonal sprouting in cortex adjacent to the infarct is part of this recovery process, but the signal that initiates axonal sprouting is not known. Growth and differentiation factor 10 (GDF10) is induced in peri-infarct neurons in mice, non-human primates and humans. GDF10 promotes axonal outgrowth in vitro in mouse, rat and human neurons through TGFβRI and TGFβRII signaling. Using pharmacogenetic gain- and loss-of-function studies, we found that GDF10 produced axonal sprouting and enhanced functional recovery after stroke; knocking down GDF10 blocked axonal sprouting and reduced recovery. RNA sequencing from peri-infarct cortical neurons revealed that GDF10 downregulated PTEN, upregulated PI3 kinase signaling and induced specific axonal guidance molecules. Using unsupervised genome-wide association analysis of the GDF10 transcriptome, we found that it was not related to neurodevelopment, but may partially overlap with other CNS injury patterns. Thus, GDF10 is a stroke-induced signal for axonal sprouting and functional recovery.

AI snapshot

Written from the paper’s full text.

TL;DR

GDF10 is induced in peri-infarct neurons after stroke across mouse, non-human primate, and human tissue and promotes axonal outgrowth via TGFβRI/II–Smad2/3 signaling. In mice, GDF10 gain of function enhances axonal sprouting and motor recovery after stroke, while GDF10 knockdown blocks sprouting and reduces recovery.

Key findings

  • GDF10 is induced in peri-infarct neurons after stroke across species: in aged rat sprouting neurons it was upregulated 1.32× by microarray and 3× by qPCR at 7 days after stroke (p<0.0009); induction was also seen in mouse, non-human primate, and human peri-infarct tissue.
  • GDF10 promotes axonal outgrowth in mouse, rat, and human neurons through TGFβRI/II and Smad2/3; 500 ng/ml produced axonal outgrowth equivalent to BDNF and FGF2, and GDF10 promoted outgrowth even in the presence of chondroitin sulfate proteoglycans.
  • In vivo, GDF10 delivery after stroke induced a robust anterior projection from motor cortex to premotor and prefrontal cortex, increased astrocytosis and angiogenesis, and improved motor recovery; GDF10 knockdown inhibited post-stroke axonal sprouting and reduced functional recovery.
  • RNA-seq from peri-infarct cortical neurons showed GDF10 downregulates PTEN, upregulates PI3K signaling, and regulates axonal guidance molecules; its transcriptome was distinct from neurodevelopment and more closely related to stroke alone than to several CNS injury or learning/memory profiles.

Methods

  • Mouse photothrombotic stroke in forelimb motor cortex; non-human primate MCA/ACA occlusion; human autopsy cases (7 stroke, 4 control); in vitro primary mouse cortical neurons, adult rat retinal ganglion cells, and human iPS-derived neurons.
  • In vivo gain/loss: GDF10 protein or GDF10 siRNA delivered via hydrogel into the stroke cavity 7 days after stroke; BDA tracing and quantitative cortical mapping of axonal connections; behavioral testing on grid walking, cylinder, and pasta handling tasks at weeks 1, 3, 5, 7, 11, and 15 after stroke.
  • RNA-seq: FACS-isolated NCAM+ cortical neurons from stroke, stroke+GDF10, control, and P4; differential expression with EdgeR at FDR<0.1; pathway analysis and unsupervised genome-wide association against 180 microarray or RNA-seq studies.

Limitations

  • The paper does not explicitly state limitations.
  • Evident from the text: human data are a retrospective convenience autopsy sample (7 stroke, 4 control) sampled at the chronic period, and the non-human primate sample is very small (2 stroke, 3 control) and sampled at 2 days, limiting cross-species and time-course conclusions.
  • Evident from the text: in vivo mouse mapping used n=8–9 per group and behavior used n=7 per group; RNA-seq compared only 3 stroke, 3 stroke+GDF10, 2 control, and P4 samples with pooling, limiting transcriptomic power.

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

Integrity signals

Registry

Crossref · 10.1038/nn.4146

Cites

57 of 57 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

59 of 104 cited sources checked · 14 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    GDF10 is one of the most highly upregulated genes during the initiation of axonal sprouting in peri-infarct cortical neurons in the aged brain.

    “Within the post-stroke axonal sprouting transcriptome, Growth and Differentiation Factor 10 (GDF10) is one of the most highly upregulated genes during the initiation of axonal sprouting in peri-infarct cortical neurons in the aged brain.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [3] Li, S. et al. An age-related sprouting transcriptome provides molecular control of axonal sprouting after stroke. Nat. Neurosci. 13, 1496–1504 (2010). source

      The Results section reports that Gdf10 was upregulated in aged sprouting neurons at day 7 after stroke, and localizes GDF10 to peri-infarct cortical neurons. It does not identify Gdf10 as one of the most highly upregulated genes; it names Igf1 and osteopontin as the most differentially regulated genes. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
  • Premisecitation

    In rodent and primate models of stroke, axonal sprouting and the formation of new connections occurs in motor, somatosensory and premotor areas.

    “In rodent and primate models of stroke, axonal sprouting and the formation of new connections occurs in motor, somatosensory and premotor areas1–4.”
    Quote in the paperSubstantial claim3 of 4 sources checked
    • cites [1] Dancause, N. et al. Extensive cortical rewiring after brain injury. J. Neurosci. 25, 10167–10179 (2005). source

      The Discussion reports novel connections between ventral premotor cortex (PMv) and somatosensory area S1 after an M1 infarct in squirrel monkeys. The section on axonal trajectories proposes sprouting but says the precise branching location can only be inferred indirectly. The paper does not itself demonstrate the claimed changes across rodent and primate models or in all three listed cortical areas; its section on other motor areas reports no significant change in PMv connectivity there. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [2] Brown, C.E., Aminoltejari, K., Erb, H., Winship, I.R. & Murphy, T.H. In vivo voltage-sensitive dye imaging in adult mice reveals that somatosensory maps lost to stroke are replaced over weeks by new structural and functional circuits with prolonged modes of activation within both the peri-infarct zone and distant sites. J. Neurosci. 29, 1719–1734 (2009). source

      The Introduction describes markers of axonal sprouting after stroke, and the neuronal-connectivity results report new connections in the reorganized forelimb area of mice. The discussion also cites prior work on axonal plasticity distant from an infarct. This paper does not establish the full claim across rodent and primate models or specifically identify new connections in premotor areas (J. Neurosci. 29:1719–1734). (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [3] Li, S. et al. An age-related sprouting transcriptome provides molecular control of axonal sprouting after stroke. Nat. Neurosci. 13, 1496–1504 (2010). source

      The Results section (“ATRX mediates axonal sprouting after stroke”; page unavailable) reports stroke-induced sprouting in motor, premotor and somatosensory areas in mice. The paper also describes new projection patterns in rats, but the passages in our copy do not establish the claim for primate stroke models. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [4] Overman, J.J. et al. A role for ephrin-A5 in axonal sprouting, recovery and activity-dependent plasticity after stroke. Proc. Natl. Acad. Sci. USA 109, E2230–E2239 (2012). source
  • Premisecitation

    GDF10 signals through TGFβ receptors.

    “and signals through TGFβ receptors (TGFβR)14,15.”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [14] Akhurst, R.J. & Hata, A. Targeting the TGFβ signalling pathway in disease. Nat. Rev. Drug Discov. 11, 790–811 (2012). source

      The TGFβ family section identifies GDFs as members of the TGFβ superfamily, and the TGFβ signalling section says members signal through type I and type II receptor complexes. The paper does not identify GDF10 specifically or establish that it uses the receptors denoted TGFβR in the citing sentence. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [15] Upadhyay, G. et al. Stem cell antigen-1 enhances tumorigenicity by disruption of growth differentiation factor-10 (GDF10)-dependent TGF-beta signaling. Proc. Natl. Acad. Sci. USA 108, 7820–7825 (2011). source
  • Premisecitation

    Stroke alone modestly but significantly induces axonal sprouting in the motor cortex adjacent to the stroke site compared to normal motor system connections.

    “As reported previously3,4,28,29, stroke alone modestly but significantly induces axonal sprouting in the motor cortex adjacent to the stroke site compared to normal motor system connections (Supplementary Fig. 6b).”
    Quote in the paperSubstantial claim3 of 4 sources checked
    • cites [3] Li, S. et al. An age-related sprouting transcriptome provides molecular control of axonal sprouting after stroke. Nat. Neurosci. 13, 1496–1504 (2010). source
    • cites [4] Overman, J.J. et al. A role for ephrin-A5 in axonal sprouting, recovery and activity-dependent plasticity after stroke. Proc. Natl. Acad. Sci. USA 109, E2230–E2239 (2012). source
    • cites [28] Clarkson, A.N. et al. Multimodal examination of structural and functional remapping in the mouse photothrombotic stroke model. J. Cereb. Blood Flow Metab. 33, 716–723 (2013). source
    • cites [29] Clarkson, A.N. et al. AMPA receptor–induced local brain-derived neurotrophic factor signaling mediates motor recovery after stroke. J. Neurosci. 31, 3766–3775 (2011). source

      The Results section, “Positive AMPAR modulation does not alter poststroke axonal sprouting” (page unavailable), says that neural repair after stroke involves new connections in the periinfarct cortex. It does not report that sprouting after stroke alone was modest but statistically significant relative to normal motor-system connections; its reported comparison is between stroke controls and CX1837-treated stroke mice. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
  • Findingcitation

    In vivo pharmacological blockade of TGFβRI/II using SB431542 and losartan decreases Smad2/3 signaling in the peri-infarct tissue during the period of GDF activity after stroke.

    “Furthermore, in vivo pharmacological blockade of TGFβRI/II using SB431542 and losartan19,20 decreases Smad2/3 signaling in the peri-infarct tissue (Supplementary Fig. 5) during the period of GDF activity after stroke.”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [19] Bar-Klein, G. et al. Losartan prevents acquired epilepsy via TGF-b signaling suppression. Ann. Neurol. 75, 864–875 (2014). source

      The Results section on local losartan application reports that losartan significantly blocked an albumin-induced increase in cortical p-Smad2/3 in vivo. The Results section on Smad phosphorylation reports an effect of SB431542 only in cultured astrocytes. This paper does not establish that either drug reduced Smad2/3 signaling in peri-infarct tissue during GDF activity after stroke; the citing sentence points to its own Supplementary Fig. 5 for that observation. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [20] Waghabi, M.C. et al. Pharmacological inhibition of transforming growth factor β signaling decreases infection and prevents heart damage in acute Chagas' Disease. Antimicrob. Agents Chemother. 53, 4694–4701 (2009). source
  • Classified as probably not a substantial claim. 46
  • May combine two claims. 41

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

    Premisecitation

    GDF10 is a divergent member of the bone morphogenetic protein (BMP)/transforming growth factor-β (TGFβ) superfamily.

    “GDF10 is a divergent member of the bone morphogenetic protein (BMP)/transforming growth factor-β (TGFβ) superfamily11–14”
    Quote in the paperNot a substantial claim2 of 4 sources checked
    • cites [11] Cunningham, N.S. et al. Growth/differentiation factor-10: a new member of the transforming growth factor-beta superfamily related to bone morphogenetic protein-3. Growth Factors 12, 99–109 (1995). source
    • cites [12] Katoh, Y. & Katoh, M. Comparative integromics on BMP/GDF family. Int. J. Mol. Med. 17, 951–955 (2006). source

      The Abstract identifies GDF10 as a BMP/GDF family gene, and the Discussion describes its conserved C-terminal TGFβ-like domain. The paper does not characterize GDF10 itself as a divergent member of the superfamily; its statement about divergence concerns primate versus mouse GDF10 promoters. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [13] Carreira, A.C. et al. Bone morphogenetic proteins: facts, challenges, and future perspectives. J. Dent. Res. 93, 335–345 (2014). source
    • cites [14] Akhurst, R.J. & Hata, A. Targeting the TGFβ signalling pathway in disease. Nat. Rev. Drug Discov. 11, 790–811 (2012). source

      The TGFβ family section identifies GDFs and BMPs as members of the TGFβ superfamily. It does not identify GDF10 specifically or describe it as divergent; searches for “GDF10” and “BMP-3b” found no such passage. Page numbers are unavailable in our copy. (LLM, conf=0.84)

      Experimental — accuracy not yet measured

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

    Premisecitation

    In humans, good functional recovery after stroke is associated with remapping of sensorimotor function in motor, somatosensory and premotor circuits and is accompanied by increases in cortical thickness in these reorganizing areas.

    “In humans, good functional recovery after stroke is associated with remapping of sensorimotor function in motor, somatosensory and premotor circuits5,6 and is accompanied by increases in cortical thickness in these reorganizing areas7.”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [5] Favre, I. et al. Upper limb recovery after stroke is associated with ipsilesional primary motor cortical activity: a meta-analysis. Stroke 45, 1077–1083 (2014). source
    • cites [6] Kantak, S.S., Stinear, J.W., Buch, E.R. & Cohen, L.G. Rewiring the brain: potential role of the premotor cortex in motor control, learning, and recovery of function following brain injury. Neurorehabil. Neural Repair 26, 282–292 (2012). source

      The section on recovery following injury to M1 describes premotor reorganization in humans after stroke, including findings in well-recovered patients. It does not establish that good recovery is associated with remapping across motor, somatosensory, and premotor circuits collectively. The cortical-thickness clause has a separate citation (7) and is not attributed to this source. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [7] Schaechter, J.D., Moore, C.I., Connell, B.D., Rosen, B.R. & Dijkhuizen, R.M. Structural and functional plasticity in the somatosensory cortex of chronic stroke patients. Brain 129, 2722–2733 (2006). source

  • May combine two claims.

    Premisecitation

    Stroke induces axonal sprouting and the formation of new connections in peri-infarct cortex that link premotor, motor, somatosensory and association areas.

    “Stroke induces axonal sprouting and the formation of new connections in peri-infarct cortex that link premotor, motor, somatosensory and association areas1–4.”
    Quote in the paperSubstantial claim3 of 4 sources checked
    • cites [1] Dancause, N. et al. Extensive cortical rewiring after brain injury. J. Neurosci. 25, 10167–10179 (2005). source

      The Discussion reports new connections between ventral premotor cortex and somatosensory area S1 after an infarct in primary motor cortex. The ‘Alteration of axonal trajectory at the lesion border’ section says sprouting near the infarct border is inferred, not directly located. The paper does not establish new connections linking all four named types of area, including association areas. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [2] Brown, C.E., Aminoltejari, K., Erb, H., Winship, I.R. & Murphy, T.H. In vivo voltage-sensitive dye imaging in adult mice reveals that somatosensory maps lost to stroke are replaced over weeks by new structural and functional circuits with prolonged modes of activation within both the peri-infarct zone and distant sites. J. Neurosci. 29, 1719–1734 (2009). source

      The Introduction (page unavailable) describes markers of axonal sprouting in peri-infarct cortex. The Figure 8 summary and the section on neuronal connectivity (pages unavailable) report increased connections from retrosplenial cortex to the reorganized forelimb area, but lost inputs from somatosensory cortex. They do not establish new connections linking all four named areas, including premotor cortex. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [3] Li, S. et al. An age-related sprouting transcriptome provides molecular control of axonal sprouting after stroke. Nat. Neurosci. 13, 1496–1504 (2010). source

      The Results section (“ATRX mediates axonal sprouting after stroke”; page unavailable) reports increased peri-infarct connections and stroke-induced sprouting in motor, premotor and somatosensory areas. The paper also describes new connection patterns, but our copy does not establish the claim’s additional link to association areas. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [4] Overman, J.J. et al. A role for ephrin-A5 in axonal sprouting, recovery and activity-dependent plasticity after stroke. Proc. Natl. Acad. Sci. USA 109, E2230–E2239 (2012). source

  • May combine two claims.

    Premiseown data

    Unlike other GDFs, GDF10 signals through TGFβRI and RII and downstream transcription factors Smad2/3, and not BMPRI/II and Smad1/5.

    “Unlike other GDFs, GDF10 signals through TGFβRI and RII and downstream transcription factors Smad2/3, and not BMPRI/II and Smad1/515”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [15] Upadhyay, G. et al. Stem cell antigen-1 enhances tumorigenicity by disruption of growth differentiation factor-10 (GDF10)-dependent TGF-beta signaling. Proc. Natl. Acad. Sci. USA 108, 7820–7825 (2011). source

      The Results (Fig. 3A, F–G) show that GDF10 requires TβRI and TβRII for Smad3 activation and induces nuclear localization of Smad2/3, but not Smad1/5. The paper does not establish that GDF10 never signals through BMP receptors or that its pathway differs from those of all other GDFs. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

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

    Premisecitation

    Compared to other GDFs, GDF10 has a unique gene structure.

    “Compared to other GDFs, GDF10 has a unique gene structure12”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [12] Katoh, Y. & Katoh, M. Comparative integromics on BMP/GDF family. Int. J. Mol. Med. 17, 951–955 (2006). source

      The Identification of the chimpanzee GDF10 gene section describes its three-exon structure, and the Discussion describes features of the GDF10 promoter. Neither section establishes that GDF10’s gene structure is unique compared with other GDFs; the paper’s promoter comparison is mainly among GDF10 orthologs. (LLM, conf=0.78)

      Experimental — accuracy not yet measured

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

    Premisecitation

    TGFβ family members stimulate astrocyte responses and modify angiogenesis and neuroinflammation.

    “TGFβ family members stimulate astrocyte responses and modify angiogenesis and neuroinflammation30,31.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [30] Smith, G.M. & Strunz, C. Growth factor and cytokine regulation of chondroitin sulfate proteoglycans by astrocytes. Glia 52, 209–218 (2005). source
    • cites [31] Wang, J. et al. Transforming growth factor β-regulated microRNA-29a promotes angiogenesis through targeting the phosphatase and tensin homolog in endothelium. J. Biol. Chem. 288, 10418–10426 (2013). source

      The Discussion reports that TGF-β signaling promotes angiogenesis by up-regulating miR-29a in endothelial cells. The paper does not address astrocyte responses or neuroinflammation (searched: astrocyte responses, neuroinflammation, brain inflammation), so it supports only the angiogenesis part of the sentence. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Study design and conductcitation

    GDF10 gain and loss of function was produced directly from the stroke through sustained release via biopolymer hydrogel, which produces release over 2-3 weeks from the stroke site without altering local inflammation, gliosis, neuronal or vascular structure.

    “To test this hypothesis, GDF10 gain and loss of function was produced directly from the stroke through sustained release via biopolymer hydrogel, which produces release over 2–3 weeks from the stroke site3,4,28 without altering local inflammation, gliosis, neuronal or vascular structure4.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [3] Li, S. et al. An age-related sprouting transcriptome provides molecular control of axonal sprouting after stroke. Nat. Neurosci. 13, 1496–1504 (2010). source

      Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured
    • cites [4] Overman, J.J. et al. A role for ephrin-A5 in axonal sprouting, recovery and activity-dependent plasticity after stroke. Proc. Natl. Acad. Sci. USA 109, E2230–E2239 (2012). source
    • cites [28] Clarkson, A.N. et al. Multimodal examination of structural and functional remapping in the mouse photothrombotic stroke model. J. Cereb. Blood Flow Metab. 33, 716–723 (2013). source

      Split into 2 atoms — 0 confirmed, 0 partially supported, 0 disputed, 0 misattributed, 2 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premiseown data

    GDF10 is induced in sprouting neurons from aged rats during the initiation phase of axonal sprouting, 7 days after stroke, 1.32× with microarray and 3× with confirmatory qPCR (p<0.0009).

    “GDF10 is induced in sprouting neurons from aged rats during the initiation phase of axonal sprouting, 7 days after stroke3: 1.32× with microarray and 3× with confirmatory qPCR (p<0.0009) at 7d after stroke3.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [3] Li, S. et al. An age-related sprouting transcriptome provides molecular control of axonal sprouting after stroke. Nat. Neurosci. 13, 1496–1504 (2010). source

      The Results sections on gene expression and mRNA confirmation report that day 7 was examined as an early stage of sprouting and that Gdf10 was upregulated in aged rat sprouting neurons in both the microarray and confirmatory qRT-PCR analyses. The passages in our copy do not give the claimed 1.32× and 3× changes or p<0.0009, so those numerical details remain unsupported. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

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

    Premisecitation

    After stroke, axonal sprouting has been demonstrated in mice, rats and non-human primates.

    “After stroke, axonal sprouting has been demonstrated in mice, rats and non-human primates1–4,10,38.”
    Quote in the paperNot a substantial claim3 of 6 sources checked
    • cites [1] Dancause, N. et al. Extensive cortical rewiring after brain injury. J. Neurosci. 25, 10167–10179 (2005). source

      The Materials and Methods and Results describe ischemic infarcts in squirrel monkeys, supporting the non-human-primate portion of the multiply cited sentence. But “Alteration of axonal trajectory at the lesion border” says that sprouting near the infarct is inferred indirectly from changed axonal trajectories, rather than directly demonstrated. This paper does not study mice or rats. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [2] Brown, C.E., Aminoltejari, K., Erb, H., Winship, I.R. & Murphy, T.H. In vivo voltage-sensitive dye imaging in adult mice reveals that somatosensory maps lost to stroke are replaced over weeks by new structural and functional circuits with prolonged modes of activation within both the peri-infarct zone and distant sites. J. Neurosci. 29, 1719–1734 (2009). source

      The Results section, “New patterns of neuronal connectivity in reorganized forelimb area” (page unavailable), reports increased axonal labeling and altered projections after stroke in mice, supporting the mouse portion of the claim. This paper does not demonstrate post-stroke axonal sprouting in rats or non-human primates; those parts of the sentence are not supported by this source. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [3] Li, S. et al. An age-related sprouting transcriptome provides molecular control of axonal sprouting after stroke. Nat. Neurosci. 13, 1496–1504 (2010). source

      The Results sections on sprouting neurons and ATRX report post-stroke axonal sprouting in rats and mice. The cited paper does not report or identify a demonstration in non-human primates. Page numbers were not provided in our copy. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [4] Overman, J.J. et al. A role for ephrin-A5 in axonal sprouting, recovery and activity-dependent plasticity after stroke. Proc. Natl. Acad. Sci. USA 109, E2230–E2239 (2012). source
    • cites [10] Wahl, A.S. et al. Neuronal repair. Asynchronous therapy restores motor control by rewiring of the rat corticospinal tract after stroke. Science 344, 1250–1255 (2014). source
    • cites [38] Ng, S.C., de la Monte, S.M., Conboy, G.L., Karns, L.R. & Fishman, M.C. Cloning of human GAP-43: growth association and ischemic resurgence. Neuron 1, 133–139 (1988). source

  • Classified as probably not a substantial claim.

    Premisecitation

    PTEN and SOCS3 inhibition induce axonal sprouting in multiple adult CNS systems, including optic nerve and spinal cord injury.

    “PTEN and SOCS3 inhibition induce axonal sprouting in multiple adult CNS systems, including optic nerve and spinal cord injury8,35.”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [8] Liu, K., Tedeschi, A., Park, K.K. & He, Z. Neuronal intrinsic mechanisms of axon regeneration. Annu. Rev. Neurosci. 34, 131–152 (2011). source
    • cites [35] Sun, F. et al. Sustained axon regeneration induced by co-deletion of PTEN and SOCS3. Nature 480, 372–375 (2011). source

      The paper reports that deleting PTEN and SOCS3 in adult retinal ganglion cells promotes axon regeneration after optic nerve injury (Abstract; no page given). It does not report PTEN/SOCS3-induced sprouting after spinal cord injury or demonstrate the effect across multiple adult CNS systems. Spinal cord injury appears only in references to other work. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

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

    Study design and conductcitation

    This experimental design has been validated by other groups conducting similar behavioral or tissue outcomes experiments.

    “This experimental design has been validated by other groups conducting similar behavioral4 or tissue outcomes experiments3,4,28,29.”
    Quote in the paperNot a substantial claim3 of 4 sources checked
    • cites [4] Overman, J.J. et al. A role for ephrin-A5 in axonal sprouting, recovery and activity-dependent plasticity after stroke. Proc. Natl. Acad. Sci. USA 109, E2230–E2239 (2012). source
    • cites [3] Li, S. et al. An age-related sprouting transcriptome provides molecular control of axonal sprouting after stroke. Nat. Neurosci. 13, 1496–1504 (2010). source
    • cites [28] Clarkson, A.N. et al. Multimodal examination of structural and functional remapping in the mouse photothrombotic stroke model. J. Cereb. Blood Flow Metab. 33, 716–723 (2013). source
    • cites [29] Clarkson, A.N. et al. AMPA receptor–induced local brain-derived neurotrophic factor signaling mediates motor recovery after stroke. J. Neurosci. 31, 3766–3775 (2011). source

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

    Study design and conductcitation

    Sample size for in vivo quantitative cortical mapping studies was estimated from previous publications with similar mechanistic studies.

    “Animal number in in vivo quantitative cortical mapping studies utilizes spatial correlation statistics, so sample size was estimated from previous publications with similar mechanistic studies3,4,28,29.”
    Quote in the paperNot a substantial claim3 of 4 sources checked
    • cites [3] Li, S. et al. An age-related sprouting transcriptome provides molecular control of axonal sprouting after stroke. Nat. Neurosci. 13, 1496–1504 (2010). source
    • cites [4] Overman, J.J. et al. A role for ephrin-A5 in axonal sprouting, recovery and activity-dependent plasticity after stroke. Proc. Natl. Acad. Sci. USA 109, E2230–E2239 (2012). source
    • cites [28] Clarkson, A.N. et al. Multimodal examination of structural and functional remapping in the mouse photothrombotic stroke model. J. Cereb. Blood Flow Metab. 33, 716–723 (2013). source

      Materials and Methods (Registration of Maps and Optical Intrinsic Signaling Imaging; page unavailable) reports five animals per group for cortical mapping. The Statistics section lists the analyses used, but neither it nor the mapping methods says that sample size was estimated from previous publications. Searched for sample-size estimation and spatial-correlation statistics. (LLM, conf=0.72)

      Experimental — accuracy not yet measured
    • cites [29] Clarkson, A.N. et al. AMPA receptor–induced local brain-derived neurotrophic factor signaling mediates motor recovery after stroke. J. Neurosci. 31, 3766–3775 (2011). source

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

    Premisecitation

    Activation of Smad2 signaling has also been reported to occur with myelin axonal growth inhibition, and direct activation of Smad2 reduces axonal outgrowth in some preparations.

    “However, activation of Smad2 signaling has also been reported to occur with myelin axonal growth inhibition, and direct activation of Smad2 reduces axonal outgrowth in some preparations22,24.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [22] Knöferle, J. et al. TGF-beta 1 enhances neurite outgrowth via regulation of proteasome function and EFABP. Neurobiol. Dis. 38, 395–404 (2010). source
    • cites [24] Stegmüller, J., Huynh, M.A., Yuan, Z., Konishi, Y. & Bonni, A. TGFbeta-Smad2 signaling regulates the Cdh1-APC/SnoN pathway of axonal morphogenesis. J. Neurosci. 28, 1961–1969 (2008). source

      The Results report basal Smad2 activation in granule neurons and show that Smad2 knockdown increases axonal length, including on myelin. They do not show that myelin activates Smad2 signaling or that directly activating Smad2 reduces axonal outgrowth; those are stronger claims than the experiments establish. No page numbers are available in our copy. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

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

    Interpretationcitation

    These growth factors may produce non-canonical signaling outside of the Smad transcription factors, such as through the MAPK pathway.

    “These growth factors may produce non-canonical signaling outside of the Smad transcription factors, such as through the MAPK pathway40,41.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [40] Yi, J.J., Barnes, A.P., Hand, R., Polleux, F. & Ehlers, M.D. TGF-beta signaling specifies axons during brain development. Cell 142, 144–157 (2010). source

      The Results section, “TGF-β Signaling is Mediated by Par6 Phosphorylation” (page unavailable), describes TGF-β receptor signaling through Par6 phosphorylation, supporting a pathway outside Smad-mediated transcription. The paper describes Par6, Smurf1 and RhoA in this pathway, but the passages found do not support MAPK as the example. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [41] Walshe, T.E., Leach, L.L. & D′Amore, P.A. TGF-beta signaling is required for maintenance of retinal ganglion cell differentiation and survival. Neuroscience 189, 123–131 (2011). source

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

    Premisecitation

    PTEN inhibition potentiates axonal sprouting in optic nerve injury and spinal cord injury, through induction of PI3kinase signaling and activation of mTOR.

    “PTEN inhibition potentiates axonal sprouting in optic nerve injury and spinal cord injury, through induction of PI3kinase signaling and activation of mTOR.”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [8] Liu, K., Tedeschi, A., Park, K.K. & He, Z. Neuronal intrinsic mechanisms of axon regeneration. Annu. Rev. Neurosci. 34, 131–152 (2011). source
    • cites [37] Zai, L. et al. Inosine alters gene expression and axonal projections in neurons contralateral to a cortical infarct and improves skilled use of the impaired limb. J. Neurosci. 29, 8187–8197 (2009). source

      The Introduction and Results discuss inosine-induced axonal sprouting after cortical stroke, while the Discussion’s “Mechanisms of action” section describes inosine activating Mst3b. Searches for PTEN, PI3kinase, and mTOR found no passage addressing whether PTEN inhibition promotes sprouting after optic nerve or spinal cord injury through that pathway. The copy therefore does not verify the attributed claim. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [38] Ng, S.C., de la Monte, S.M., Conboy, G.L., Karns, L.R. & Fishman, M.C. Cloning of human GAP-43: growth association and ischemic resurgence. Neuron 1, 133–139 (1988). source

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

    Findingcitation

    With simple growth factor withdrawal for three weeks, most iPS-NPCs develop into neurons expressing Tuj1 or MAP2.

    “With simple growth factor withdrawal for three weeks, most of these iPS-NPCs develop into neurons expressing Tuj1 or MAP250”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [50] Andres, R.H. et al. Human neural stem cells enhance structural plasticity and axonal transport in the ischaemic brain. Brain 134, 1777–1789 (2011). source

      The Materials and methods section identifies the studied NPCs as derived from foetal cortical tissue, not induced pluripotent stem cells. The Results section reports that most transplanted NPCs remained immature after five weeks in the rat brain, but does not test three weeks of growth-factor withdrawal. Searches for iPS-NPC differentiation under that condition found no passage addressing the claim; the different experimental condition is not a contradiction. (LLM, conf=0.78)

      Experimental — accuracy not yet measured

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

    Premisecitation

    Such context-specific signaling is a hallmark of TGFβ in its role in cancer, where it switches from inhibition of tumor growth and metastasis to promotion of endothelial to mesenchymal transition and metastasis as the local tumor environment evolves.

    “Such context-specific signaling is a hallmark of TGFβ in its role in cancer, where it switches from inhibition of tumor growth and metastasis to promotion of endothelial to mesenchymal transition and metastasis as the local tumor environment evolves14,43.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [14] Akhurst, R.J. & Hata, A. Targeting the TGFβ signalling pathway in disease. Nat. Rev. Drug Discov. 11, 790–811 (2012). source

      Split into 2 atoms — 1 confirmed, 1 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured
    • cites [43] Bahjat, F.R. et al. Proof of concept: pharmacological preconditioning with a Toll-like receptor agonist protects against cerebrovascular injury in a primate model of stroke. J. Cereb. Blood Flow Metab. 31, 1229–1242 (2011). source

      Split into 2 atoms — 0 confirmed, 0 partially supported, 0 disputed, 0 misattributed, 2 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured

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

    Findingcitation

    The iPS-NPC differentiation approach generates neurons with markers of maturity by transcript profiling, immunohistochemical staining and action potential generation.

    “This approach generates neurons with markers of maturity by transcript profiling, immunohistochemical staining and action potential generation (Fig. 1, ref 27).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [27] Paşca, S.P. et al. Using iPSC-derived neurons to uncover cellular phenotypes associated with Timothy syndrome. Nat. Med. 17, 1657–1662 (2011). source

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

    Premisecitation

    BDNF and NT3 treatment promotes further neuronal differentiation.

    “BDNF and NT3 treatment were used to promote further neuronal differentiation27”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [27] Paşca, S.P. et al. Using iPSC-derived neurons to uncover cellular phenotypes associated with Timothy syndrome. Nat. Med. 17, 1657–1662 (2011). source

      The Neural differentiation protocol passage (section labeled Abstract; page unavailable) describes differentiation into neurons but does not mention BDNF or NT3 treatment. Searches for BDNF, NT3, and their expanded names found no passage in our copy establishing that either treatment was used to promote further differentiation. (LLM, conf=0.72)

      Experimental — accuracy not yet measured

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

    Findingcitation

    This iPS-NPC line is highly similar to other iPS-NPCs and ES-NPCs derived under standard conditions (Pearson correlation 0.926-0.959).

    “Clustering and Pearson analysis49 demonstrate that this iPS-NPCs is highly similar to other iPS-NPCs and ES-NPCS derived under standard conditions (Pearson: 0.926–0.959)”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [49] Patterson, M. et al. let-7 miRNAs can act through notch to regulate human gliogenesis. Stem Cell Reports 3, 758–773 (2014). source

      The Cell Culture section (page unavailable) describes deriving neural progenitors from pluripotent stem-cell lines, but does not compare the citing paper’s iPS-NPC line with other iPS-NPCs or ES-NPCs. The Results section reports a Pearson comparison between let-7-treated PSC-NPCs and tissue-derived NPCs, not the claimed comparison. Searches for the 0.926–0.959 range and the iPS-NPC/ES-NPC clustering comparison found no supporting passage in our copy. (LLM, conf=0.77)

      Experimental — accuracy not yet measured

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

    Premisecitation

    Inhibition of SOCS3 further promotes axonal sprouting on top of PTEN inhibition.

    “Inhibition of SOCS3 further promotes axonal sprouting on top of PTEN inhibition8,37,38.”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [8] Liu, K., Tedeschi, A., Park, K.K. & He, Z. Neuronal intrinsic mechanisms of axon regeneration. Annu. Rev. Neurosci. 34, 131–152 (2011). source
    • cites [37] Zai, L. et al. Inosine alters gene expression and axonal projections in neurons contralateral to a cortical infarct and improves skilled use of the impaired limb. J. Neurosci. 29, 8187–8197 (2009). source

      The Results section reports increased corticospinal axon sprouting after inosine treatment, and the Discussion’s “Mechanisms of action” section describes inosine and Mst3b. Neither passage addresses SOCS3 inhibition, PTEN inhibition, or their combined effect. Searches for SOCS3/PTEN and their expanded names found no passage supporting the attributed claim; page numbers were not available in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [38] Ng, S.C., de la Monte, S.M., Conboy, G.L., Karns, L.R. & Fishman, M.C. Cloning of human GAP-43: growth association and ischemic resurgence. Neuron 1, 133–139 (1988). source

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

    Premisecitation

    In other in vitro systems, TGFβ promotes axonal outgrowth of injured neurons and blockade of TGFβRII alters neuronal polarity and leads to shorter axons.

    “In other in vitro systems, TGFβ promotes axonal outgrowth of injured neurons21,22,39,40 and blockade of TGFβRII alters neuronal polarity and leads to shorter axons42.”
    Quote in the paperNot a substantial claim2 of 5 sources checked
    • cites [21] Ishihara, A., Saito, H. & Abe, K. Transforming growth factor-beta 1 and -beta 2 promote neurite sprouting and elongation of cultured rat hippocampal neurons. Brain Res. 639, 21–25 (1994). source
    • cites [22] Knöferle, J. et al. TGF-beta 1 enhances neurite outgrowth via regulation of proteasome function and EFABP. Neurobiol. Dis. 38, 395–404 (2010). source
    • cites [39] Abe, K., Chu, P.J., Ishihara, A. & Saito, H. Transforming growth factor-beta 1 promotes re-elongation of injured axons of cultured rat hippocampal neurons. Brain Res. 723, 206–209 (1996). source
    • cites [40] Yi, J.J., Barnes, A.P., Hand, R., Polleux, F. & Ehlers, M.D. TGF-beta signaling specifies axons during brain development. Cell 142, 144–157 (2010). source

      The Results section, “Axon Specification by TGF-β Signaling is Cell Autonomous” (page unavailable), shows that blocking TβR2 signaling in cultured neurons disrupted polarity and reduced the length of their longest neurite. It does not establish that the affected neurons had shorter axons: most lacked a tau-1-positive axon, and the short processes were described as neurites. The injured-neuron clause is attached to other citations, not this one. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [42] Lenferink, A.E. et al. Transcriptome profiling of a TGF-beta-induced epithelial-to-mesenchymal transition reveals extracellular clusterin as a target for therapeutic antibodies. Oncogene 29, 831–844 (2010). source

      The Introduction describes TGF-β signaling, and the section on BRI-JM01 cells examines epithelial-to-mesenchymal transition in a mammary tumor cell line. Searches for injured neurons, axonal outgrowth, neuronal polarity, and TGF-βRII blockade found no passage addressing either neuronal claim; the cited paper cannot verify them. (LLM, conf=0.77)

      Experimental — accuracy not yet measured

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

    Findingcitation

    This iPS-NPC line is relatively homogenous by immunostaining.

    “and by immunostaining are relatively homogenous49”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [49] Patterson, M. et al. let-7 miRNAs can act through notch to regulate human gliogenesis. Stem Cell Reports 3, 758–773 (2014). source

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

    Premisecitation

    TGFβ promotes tissue fibrosis and has potential direct deleterious effects on kidney, liver and immune function.

    “TGFβ promotes tissue fibrosis and has potential direct deleterious effects on kidney, liver and immune function14.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [14] Akhurst, R.J. & Hata, A. Targeting the TGFβ signalling pathway in disease. Nat. Rev. Drug Discov. 11, 790–811 (2012). source

  • May combine two claims.

    Findingown data

    GDF10 produces axonal sprouting and enhanced functional recovery after stroke.

    “GDF10 produces axonal sprouting and enhanced functional recovery after stroke”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Knocking down GDF10 blocks axonal sprouting and reduces recovery.

    “knocking down GDF10 blocks axonal sprouting and reduces recovery.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Unsupervised genome-wide association analysis of the GDF10 transcriptome shows that it is not related to neurodevelopment but may partially overlap with other CNS injury patterns.

    “Unsupervised genome-wide association analysis of the GDF10 transcriptome shows that it is not related to neurodevelopment but may partially overlap with other CNS injury patterns.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    RNA-seq from peri-infarct cortical neurons indicates that GDF10 downregulates PTEN and upregulates PI3 kinase signaling and induces specific axonal guidance molecules.

    “RNA-seq from peri-infarct cortical neurons indicates that GDF10 downregulates PTEN and upregulates PI3 kinase signaling and induces specific axonal guidance molecules.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    This is a non-isotropic increase in projections, with a unique frontally-projecting axonal connection with GDF10 delivery after stroke.

    “This is a non-isotropic increase in projections, with a unique frontally-projecting axonal connection with GDF10 delivery after stroke.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    GDF10 siRNA significantly knocks down GDF10 protein levels and inhibits axonal outgrowth from cortical neurons.

    “GDF10 siRNA significantly knocks down GDF10 protein levels (Supplementary Figs. 3a,c) and inhibits axonal outgrowth from cortical neurons (Fig. 2b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In mouse primary cortical neurons, GDF10 produces an increase in axonal outgrowth, with the dose of 500ηg/ml producing equivalent axonal outgrowth to similar doses of BDNF and FGF2.

    “In mouse primary cortical neurons, GDF10 produces an increase in axonal outgrowth, with the dose of 500ηg/ml producing equivalent axonal outgrowth to similar doses of the well-described axonal sprouting growth factors braid derived growth factor (BDNF) and fibroblast growth factor 2 (FGF2) (Fig. 2a).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In vivo, GDF10 enhances axonal sprouting in peri-infarct cortex and improves motor recovery after stroke.

    “In vivo, GDF10 enhances axonal sprouting in peri-infarct cortex and improves motor recovery after stroke”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    GDF10 significantly increased astrocytosis, as indicated by an increase in the area of GFAP+ processes, and in blood vessel area, as indicated by an increase in PECAM+ endothelial cell area, above that seen in stroke, with no significant effect on microglial staining.

    “GDF10 significantly increased astrocytosis, as indicated by an increase in the area of GFAP+ processes, and in blood vessel area, as indicated by an increase in PECAM+ endothelial cell area, above that seen in stroke, with no significant effect on microglial staining (Fig. 5).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    GDF10 is induced in rodents (7 days after stroke), non-human primates (2 days) and humans (chronic period).

    “In these studies, GDF10 is induced in rodents (7 days after stroke), non-human primates (2 days) and humans (chronic period) indicating a large range of species and a broad time period for upregulation after stroke.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    GDF10 immunoreactivity within peri-infarct cortex is not present in cells with markers of microglia and astrocytes, but is seen in the extracellular matrix.

    “GDF10 immunoreactivity within peri-infarct cortex is not present in cells with markers of microglia and astrocytes, but is seen in the extracellular matrix, as expected of a secreted protein in the peri-infarct cortex (Fig. 1, Supplementary Fig. 2d,e).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    After stroke, GDF10 is induced in cells with markers of neurons and in brain tissue surrounding these neurons.

    “After stroke, GDF10 is induced in cells with markers of neurons and in brain tissue surrounding these neurons (Fig. 1, Supplementary Fig. 2a,b,d).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Knockdown of GDF10 blocks axonal sprouting and behavioral recovery.

    “knockdown of GDF10 blocks this axonal sprouting and behavioral recovery.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    GDF10 enhances axonal outgrowth and knockdown of GDF10 blocks axonal outgrowth in several neuronal types, with a dose effect similar to other growth promoting factors.

    “These studies show that GDF10 enhances axonal outgrowth and knockdown of GDF10 blocks axonal outgrowth in several neuronal types, with a dose effect that is similar to other growth promoting factors.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The axonal outgrowth effect of TGFβ is weaker than that of GDF10, as well as BDNF, and has a ceiling effect.

    “However, the axonal outgrowth effect of TGFβ is weaker than that of GDF10, as well as BDNF, and has a ceiling effect (Fig. 3g).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Stroke+GDF10 induces a statistically significant change in cortical connections compared to stroke alone and stroke+protein control, with a robust projection from motor cortex anteriorly to premotor and prefrontal cortex.

    “Quantitative connectional mapping shows that stroke+GDF10 induces a statistically significant change in cortical connections compared to stroke alone and stroke+protein control, with a robust projection from motor cortex anteriorly to premotor and prefrontal cortex (n = 8–9 per group, Fig. 4a–c).”
    Quote in the paperSubstantial claim

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

    Findingreasoning

    There is a variance in statistical spread across transcriptomes, even from the same cell population in the same experiment.

    “There is a variance in statistical spread across transcriptomes, even from the same cell population in the same experiment (Fig. 8c), which likely has to do with the fact that these transcriptional profiles are mostly from specific cell types, rather than from large in vitro preparations or tissue-level sources.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    PTEN canonical signaling (8 genes in class) is downregulated, and PI3K canonical signaling (12 genes in class) is upregulated with GDF10 in stroke.

    “PTEN canonical signaling (8 genes in class) is downregulated, and PI3K canonical signaling (12 genes in class) is upregulated with GDF10 in stroke (Supplementary Tables 3–6).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Knockdown of GDF10 after stroke abolishes the normal process of post-stroke axonal sprouting and reduces functional recovery.

    “Further, knockdown of GDF10 after stroke abolishes the normal process of post-stroke axonal sprouting and reduces functional recovery.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingnone given

    GDF10 activates a coordinated upregulation of genes in the PI3kinase pathway, downregulation of genes in the PTEN pathway and control of specific axonal guidance molecules, including a downregulation of SOCS3.

    “GDF10 activates a coordinated upregulation of genes in the PI3kinase pathway, downregulation of genes in the PTEN pathway and control of specific axonal guidance molecules, including a downregulation of SOCS3.”
    Quote in the paperSubstantial claim

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

    Findingown data

    Between 67 and 85% of the reads mapped uniquely to the mouse genome.

    “Between 67 and 85% of the reads mapped uniquely to the mouse genome.”
    Quote in the paperNot a substantial claim

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

    Findingown data

    Across the samples, more than 25% of the annotated genes were detected by at least 50 reads.

    “Across the samples >25% of the annotated genes have been detected by at least 50 reads.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Many studies have identified molecules that block axonal sprouting after CNS injury, such as myelin proteins or chondroitin sulfate proteoglycans.

    “There have been many studies of the molecules that block axonal sprouting after CNS injury, such as myelin proteins or chondroitin sulfate proteoglycans (CSPGs)8–10”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [8] Liu, K., Tedeschi, A., Park, K.K. & He, Z. Neuronal intrinsic mechanisms of axon regeneration. Annu. Rev. Neurosci. 34, 131–152 (2011). source
    • cites [9] Soleman, S., Filippov, M.A., Dityatev, A. & Fawcett, J.W. Targeting the neural extracellular matrix in neurological disorders. Neuroscience 253, 194–213 (2013). source
    • cites [10] Wahl, A.S. et al. Neuronal repair. Asynchronous therapy restores motor control by rewiring of the rat corticospinal tract after stroke. Science 344, 1250–1255 (2014). source

  • May combine two claims.

    Study design and conductown data

    There is no significant difference in BDA injection location and volumes, or in infarct volumes across all experimental conditions.

    “There is no significant difference in BDA injection location and volumes, or in infarct volumes across all experimental conditions (Supplementary Fig. 7a,b).”
    Quote in the paperSubstantial claim

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

    Premisecitation

    GDF10 mRNA is strongly expressed in the developing brain.

    “Though GDF10 mRNA is strongly expressed in the developing brain16,17”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [16] Söderström, S. & Ebendal, T. Localized expression of BMP and GDF mRNA in the rodent brain. J. Neurosci. Res. 56, 482–492 (1999). source

      Not checked in the free run.

    • cites [17] Zhao, R., Lawler, A.M. & Lee, S.J. Characterization of GDF-10 expression patterns and null mice. Dev. Biol. 212, 68–79 (1999). source

  • Classified as probably not a substantial claim.

    Premisecitation

    Human and rodent cells may respond differently to TGFβ/BMP signaling.

    “Human and rodent cells may respond differently to TGFβ/BMP signaling25.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [25] Vallier, L. & Pedersen, R.A. Human embryonic stem cells: an in vitro model to study mechanisms controlling pluripotency in early mammalian development. Stem Cell Rev. 1, 119–130 (2005). source

  • May combine two claims.

    Interpretationown data

    GDF10 induces substantial axonal sprouting and synapse formation after stroke into premotor and prefrontal areas, and blocking GDF10 eliminates the normal pattern of axonal sprouting seen in stroke to produce a pattern of motor system connections that is not statistically different from that of the normal brain.

    “These data show that GDF10 induces substantial axonal sprouting and synapse formation after stroke into premotor and prefrontal areas, and blocking GDF10 eliminates the normal pattern of axonal sprouting seen in stroke to produce a pattern of motor system connections that is not statistically different from that of the normal brain.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    These data provide independent confirmation that Smad2 downstream signaling from TGFβRI/II promotes axonal sprouting, but suggest that additional elements in GDF10 action play a role in the full axonal sprouting effect.

    “These data provide independent confirmation that Smad2 downstream signaling from TGFβRI/II promotes axonal sprouting, but suggest that additional elements in GDF10 action play a role in the full axonal sprouting effect.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    In humans, the initial cloning studies of GAP43 showed that it is induced in peri-infarct cortex after stroke and increases cortical thickness in regions of cortical map plasticity during recovery.

    “In humans, the initial cloning studies of GAP43, the paradigmatic neuronal growth cone marker, showed that it is induced in peri-infarct cortex after stroke39 and increases cortical thickness in regions of cortical map plasticity during recovery7.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [39] Abe, K., Chu, P.J., Ishihara, A. & Saito, H. Transforming growth factor-beta 1 promotes re-elongation of injured axons of cultured rat hippocampal neurons. Brain Res. 723, 206–209 (1996). source
    • cites [7] Schaechter, J.D., Moore, C.I., Connell, B.D., Rosen, B.R. & Dijkhuizen, R.M. Structural and functional plasticity in the somatosensory cortex of chronic stroke patients. Brain 129, 2722–2733 (2006). source

  • May combine two claims.

    Interpretationown data

    Administration of GDF10 protein improves motor recovery after stroke and knocking down GDF10 levels significantly decreases functional recovery.

    “Thus, administration of GDF10 protein improves motor recovery after stroke and knocking down GDF10 levels significantly decreases functional recovery.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    The present data indicate that GDF10 is induced after stroke in a wide range of species, promotes axonal outgrowth in vitro in human, mouse and rat neurons, and induces axonal sprouting and functional recovery after stroke in vivo.

    “The present data indicate that GDF10 is induced after stroke in a wide range of species, promotes axonal outgrowth in vitro in human, mouse and rat neurons, and induces axonal sprouting and functional recovery after stroke in vivo.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    GDF10 delivery not only enhances axonal sprouting, but also induces vascular remodeling and astrocytosis.

    “These data indicate that GDF10 delivery not only enhances axonal sprouting, but also induces vascular remodeling and astrocytosis.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    GDF10 delivery in stroke, which promotes axonal sprouting in the adult, regulates a dramatically different and smaller set of genes from that seen during the initial process of axonal sprouting in neurodevelopment and in the adult state, GDF10 induces a unique transcriptome after stroke.

    “These results indicate that GDF10 delivery in stroke, which promotes axonal sprouting in the adult, regulates a dramatically different and smaller set of genes from that seen during the initial process of axonal sprouting in neurodevelopment and in the adult state, GDF10 induces a unique transcriptome after stroke.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    GDF10 is a post-stroke axonal sprouting signal that induces functional recovery.

    “These findings suggest that GDF10 is a post-stroke axonal sprouting signal that induces functional recovery.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Axonal sprouting in the adult occurs in a limited manner after stroke, optic nerve lesions, spinal cord injury and in models of neurodegenerative diseases.

    “Axonal sprouting in the adult occurs in a limited manner after stroke, optic nerve lesions, spinal cord injury and in models of neurodegenerative diseases8,9.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [8] Liu, K., Tedeschi, A., Park, K.K. & He, Z. Neuronal intrinsic mechanisms of axon regeneration. Annu. Rev. Neurosci. 34, 131–152 (2011). source
    • cites [9] Soleman, S., Filippov, M.A., Dityatev, A. & Fawcett, J.W. Targeting the neural extracellular matrix in neurological disorders. Neuroscience 253, 194–213 (2013). source

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

    Premisecitation

    Much scientific study has focused on the molecular components of the injury response that block axonal sprouting, such as the glial growth inhibitors NogoA and the chondroitin sulfate proteoglycans.

    “Much scientific study has focused on the molecular components of the injury response that block axonal sprouting, such as the glial growth inhibitors NogoA and the chondroitin sulfate proteoglycans8–10.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [8] Liu, K., Tedeschi, A., Park, K.K. & He, Z. Neuronal intrinsic mechanisms of axon regeneration. Annu. Rev. Neurosci. 34, 131–152 (2011). source
    • cites [9] Soleman, S., Filippov, M.A., Dityatev, A. & Fawcett, J.W. Targeting the neural extracellular matrix in neurological disorders. Neuroscience 253, 194–213 (2013). source
    • cites [10] Wahl, A.S. et al. Neuronal repair. Asynchronous therapy restores motor control by rewiring of the rat corticospinal tract after stroke. Science 344, 1250–1255 (2014). source

  • May combine two claims.

    Interpretationown data

    These behavioral data indicate that endogenous GDF10 has an important role in normal recovery after stroke and that the neuronal sprouting mapped in vivo may represent formation of neural circuits that cause behavioral improvement.

    “These behavioral data indicate that endogenous GDF10 has an important role in normal recovery after stroke and that the neuronal sprouting mapped in vivo may represent formation of neural circuits that cause behavioral improvement.”
    Quote in the paperSubstantial claim

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

    Interpretationreasoning

    These discrepant findings of both axonal sprouting and axonal growth inhibition with TGFβ may stem from non-canonical signaling or context-specific signaling.

    “These discrepant findings of both axonal sprouting and axonal growth inhibition with TGFβ may stem from non-canonical signaling or context-specific signaling.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Behaviors were scored by observers masked to the treatment group of the animals.

    “Behaviors were scored by observers who were masked to the treatment group of the animals.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Interpretationnone given

    Systemic delivery of TGFβ would activate its signaling systems in all tissue of the body.

    “With such a therapy in mind, systemic delivery of TGFβ would activate its signaling systems in all tissue of the body.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductown data

    Protein knockdown of TβRII, Smad2, Smad3 and GDF10 by siRNA was evaluated with Western blotting.

    “TβRII, Smad2, Smad3 and GDF10 siRNA protein knockdown was evaluated with Western blotting (Supplementary Fig. 3c,d).”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    APC sort gates were set using positive and negative controls prior to neuron sorting.

    “APC sort gates were set using positive and negative controls prior to neuron sorting.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Scanning parameters were kept constant across treatment conditions for quantification of GFAP, PECAM and IBA1 immunoreactive areas.

    “The parameters for scanning were kept constant across treatment conditions.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Following alignment and read quantification, quality control was performed using sample clustering, consistency of replicates, and average gene coverage.

    “Following alignment and read quantification, we performed quality control using a variety of indices, including sample clustering, consistency of replicates, and average gene coverage.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    RNA quality was verified with RIN>7 on an Agilent Bioanalyzer.

    “RNA quality was verified (RIN>7) on an Agilent Bioanalyzer.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    In all experiments, no-primary and no-secondary antibody controls were run in parallel.

    “In all experiments, no-primary and no-secondary antibody controls were run in parallel.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Sample size in behavioral studies was assessed by power analysis using a significance level of α=0.05 with 80% power to detect differences in ANOVA.

    “Sample size in in behavioral studies was assessed by power analysis using a significance level of α=0.05 with 80% power to detect differences in ANOVA.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    There was no specific staining with the no-primary and no-secondary antibody controls.

    “There was no specific staining with these controls.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    The action of TGFβRI/II signaling in axonal outgrowth may be context specific in terms of the local tissue environment and differ between local axonal sprouting in peri-infarct cortex and distant axonal sprouting from corticospinal neurons after spinal cord injury.

    “The action of TGFβRI/II signaling in axonal outgrowth may be context specific in terms of the local tissue environment and differ between local axonal sprouting in peri-infarct cortex and distant axonal sprouting from corticospinal neurons after spinal cord injury.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    One control sample failed QC and was excluded from analysis.

    “One control sample failed QC and was excluded from analysis.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    BDA-positive processes were marked by an observer blind to the treatment conditions.

    “BDA-positive processes were marked in x,y coordinates relative to the center of the injection site by an observer blind to the treatment conditions.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    The observer was blinded to the treatment conditions in each well of the culture plate.

    “The observer was blinded to the treatment conditions in each well of the culture plate.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Study design and conductown data

    The size and location of each BDA injection and stroke size did not vary significantly across individuals or by treatment condition.

    “The size and location of each BDA injection and stroke size did not vary significantly across individuals or by treatment condition (Supplementary Fig. 7).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    GDF10 represents a significant regulator of axonal outgrowth and functional recovery after stroke and may present a valuable target for therapeutic intervention.

    “In all, GDF10 represents a significant regulator of axonal outgrowth and functional recovery after stroke and may present a valuable target for therapeutic intervention.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    No animals were excluded from analyses.

    “No animals were excluded from analyses.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Mice were randomly allocated to treatment condition.

    “Mice were randomly allocated to treatment condition”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    All results were analyzed with the investigator blinded to treatment condition.

    “all results were analyzed with the investigator blinded to treatment condition.”
    Quote in the paperNot a substantial claim
81 of 81

  • The paper relies on this without support.

    The factor(s) triggered by stroke to promote the initiation of a molecular growth program and axonal sprouting are unknown.

    “but the factor(s) that are triggered by stroke to promote the initiation of a molecular growth program and axonal sprouting are unknown.”

  • The paper relies on this without support.

    A role for GDF10 in the adult brain or after CNS injury has not been described.

    “a role for GDF10 in the adult brain or after CNS injury has not been described.”

  • The paper relies on this without support.

    A role for GDF10 in axonal outgrowth has not been previously established.

    “A role for GDF10 in axonal outgrowth has not been previously established.”

  • The paper relies on this without support.

    The GDF10 siRNA combination that produced the greatest knockdown in vitro also effectively knocks down GDF10 in vivo.

    “The combination of MSS236597 and MSS236598 siRNA produced the greatest knockdown in GDF10 protein expression (Supplementary Fig. 3c).”
    “This GDF10 siRNA (n = 8) or scrambled control siRNA (n = 8) duplex with RNAiMAX (cat. no. 13778-075, Invitrogen) (6 μl) was introduced at 150 nM directly into the stroke cavity”
4 of 4

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.

  • BDA tracer injection is dated both 21 and 28 days after stroke

    Minor

    The Results place treatment 7 days after stroke and tracer injection 21 days later, or day 28. The Methods place tracer injection at day 21 after stroke, 14 days after treatment. This changes the interval used to assess axonal sprouting.

  • RNA-seq read lengths conflict within the Methods

    Minor

    The Methods describe the RNA-seq as paired-end 2 × 69 reads, then describe the HiSeq 2000 run as paired-end 2 × 100 bp. Both cannot be the read length of the same sequencing run as described.

2 of 2

Cites

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Authors
Li, S., Nie, E. H., Yin, Y., Benowitz, L. I., Tung, S., Vinters, H. V., Bahjat, F. R., Stenzel-Poore, M. P., Kawaguchi, R., Coppola, G., & Carmichael, S. T.
Affiliations
  • University of California, Los Angeles (US)
  • Boston Children's Hospital (US)
  • Oregon Health & Science University (US)
Corresponding author
Carmichael, S Thomas
ORCID
Venue
Nature Neuroscience
Publisher
Springer Science and Business Media LLC
Published
2015-10-26
Volume
18
Issue
12
Pages
1737-1745
ISSN
  • 1097-6256 (ISSN-L)
  • 1546-1726
DOI
10.1038/nn.4146
PMC
PMC4790086
OpenAlex
W2273229061
Semantic Scholar
c476ee7ddb6be5fa6b51c8ae52ed69c7e4cd242a
Cited by
181
References
57
Citations by year
2026: 11 · 2025: 13 · 2024: 22 · 2023: 16 · 2022: 11 · 2021: 17 · 2020: 22 · 2019: 25 · 2018: 13 · 2017: 18 · 2016: 10 · 2015: 3
FWCI
6.05
Type
journal article
Language
en
Open access
Licence
www.springer.com/tdm (tdm)
Locations
  • Nature Neuroscience (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
  • American Heart Association
  • Dr. Miriam and Sheldon G. Adelson Medical Research Foundation
  • National Institutes of Health (P51-OD011092)
  • National Institute of Neurological Disorders and Stroke (R01 NS085019, U01 NS064953, P30NS062691, NS086431, F31 NS086431, NS085019)
Topics
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Nerve injury and regeneration
  • Health Sciences › Medicine › Rehabilitation › Stroke Rehabilitation and Recovery
  • Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
Keywords
GDF10, stroke, axonal sprouting, functional recovery, TGF-beta signaling, TGF-beta type I receptor, TGFBR2, PTEN, PI3K signaling, neuroplasticity, central nervous system injury
MeSH terms
Aged, Aged, 80 and over, Animals, Animals, Newborn, Axons, Humans, Macaca mulatta, Mice, Inbred C57BL, Middle Aged, Retrospective Studies, Recovery of Function, Stroke, Mice, Growth Differentiation Factor 10

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-18.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-18.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

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