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Cited by 280Autism Spectrum Disorder ResearchGenetics and Neurodevelopmental DisordersNeuroscience and Neuropharmacology Research

Regulation of autism-relevant behaviors by cerebellar-prefrontal cortical circuits

Open access
Kelly, Elyza, Meng, Fantao, Fujita, Hirofumi, Morgado, Felipe, Kazemi, Yasaman, Rice, Laura C., Ren, Chongyu, Escamilla, Christine Ochoa +23 more·2020·Nature Neuroscience
DOI 10.1038/s41593-020-0665-zPMC PMC7483861
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Abstract

Cerebellar dysfunction has been demonstrated in autism spectrum disorders (ASDs); however, the circuits underlying cerebellar contributions to ASD-relevant behaviors remain unknown. In this study, we demonstrated functional connectivity between the cerebellum and the medial prefrontal cortex (mPFC) in mice; showed that the mPFC mediates cerebellum-regulated social and repetitive/inflexible behaviors; and showed disruptions in connectivity between these regions in multiple mouse models of ASD-linked genes and in individuals with ASD. We delineated a circuit from cerebellar cortical areas Right crus 1 (Rcrus1) and posterior vermis through the cerebellar nuclei and ventromedial thalamus and culminating in the mPFC. Modulation of this circuit induced social deficits and repetitive behaviors, whereas activation of Purkinje cells (PCs) in Rcrus1 and posterior vermis improved social preference impairments and repetitive/inflexible behaviors, respectively, in male PC-Tsc1 mutant mice. These data raise the possibility that these circuits might provide neuromodulatory targets for the treatment of ASD. Kelly et al. describe two cerebellum–thalamus–mPFC pathways in mice that regulate social and repetitive behavior. PC activation in Rcrus1 and posterior vermis improved social and reduced repetitive behaviors, respectively, in PC-Tsc1 mutant mice.

AI snapshot

Written from the paper’s full text.

TL;DR

This study delineates cerebellar–prefrontal circuits in mice through which Rcrus1 and posterior vermis Purkinje cell output converges on ventromedial thalamus–medial prefrontal cortex projections to regulate autism-relevant social and repetitive/inflexible behaviors. It also reports disrupted cerebellar–mPFC connectivity in multiple ASD-linked mouse models and in individuals with ASD, and shows that circuit modulation can rescue behaviors in adult PC-Tsc1 mutant mice.

Key findings

  • Cerebellar Purkinje cell dysfunction or chemogenetic inhibition of Rcrus1 or posterior vermis increased left prelimbic mPFC single-unit firing; inhibiting left mPFC rescued social deficits and repetitive grooming in PC-Tsc1 mutant mice.
  • A polysynaptic circuit was delineated from Rcrus1 and posterior vermis through cerebellar nuclei (lateral nucleus and medial/fastigial nucleus) and ventromedial thalamus to left prelimbic mPFC; optogenetic activation of VM thalamic terminals in mPFC induced social deficits and repetitive grooming, while optogenetic silencing rescued both in PC-Tsc1 mutants.
  • Distinct cerebellar domains regulated different behaviors: Rcrus1/lateral nucleus modulation rescued social behaviors but not repetitive behaviors, whereas posterior vermis Purkinje cell stimulation selectively reduced repetitive grooming and improved water Y-maze flexibility without improving social preference in PC-Tsc1 mutants.
  • Connectivity was disrupted in ASD models and humans: structural covariance between Rcrus1 and left PRL mPFC was altered across 94 ASD-linked mouse lines (FDR q = 0.0191), with 30 of 94 models showing significant volume changes (binomial P = 2.02 × 10−18; combined q < 0.0001); human POND structural covariance showed Rcrus1–left rectus gyrus disruption (P = 0.012) that did not survive multiple corrections (q > 0.05); human resting-state fMRI in 8 ASD vs 8 TD males showed mPFC/Rcrus1–Rcrus2 alterations (P < 0.001, FDR q < 0.05) and reduced seed-to-voxel functional connectivity (P < 0.001, k > 50).

Methods

  • Mice: PC-Tsc1 mutants (PCCre;Tsc1flox/flox) and littermate controls, aged 5–12 weeks; only male animals were used for behavioral experiments and electrophysiological recordings. Methods included stereotaxic viral injections, chemogenetic and optogenetic modulation, in vivo single-unit recordings, acute slice electrophysiology, viral tracing, and behavioral assays (three-chamber social approach/novelty, social olfaction, grooming, water T/Y-maze reversal, open field, elevated plus maze, rotarod).
  • MRI/connectivity: mouse structural covariance MRI across 94 ASD-linked gene models (2,414 mice, 67.7% male, PND 30–245), plus a subset of 30 models (n = 559 mutants, n = 1,101 controls); human structural MRI from POND and other data (902 scans, 425 ASD and 477 control, from 648 individuals, age 2–50 years, mean 13); human resting-state fMRI in 8 TD male adults (mean age 20.75 ± 1.91 years) and 8 ASD male adults (mean age 26.13 ± 9.79 years).
  • Analysis: structural covariance modeled with sex and sex interaction covariates, two-sided t-tests, Benjamini–Hochberg FDR q values; behavioral data analyzed with two- or three-way ANOVA and Sidak post hoc tests; electrophysiology with t-tests or Mann–Whitney tests; sample size estimated with PASS 12.

Limitations

  • Stated: the human structural covariance finding between Rcrus1 and left rectus gyrus did not survive multiple corrections (q > 0.05); the human resting-state fMRI cohort was small; the precise function of these regions/circuits during social interactions and the broader cerebellar–cerebro cortical network map require future research; additional cortical and subcortical regions/routes may contribute; mPFC is a less accessible target for noninvasive neuromodulation.
  • Evident from the text: behavioral and electrophysiological experiments used only male mice, limiting generalization to females; mice were of mixed genetic backgrounds; some human imaging analyses involved small samples or did not survive correction.

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

Integrity signals

Registry

Crossref · 10.1038/s41593-020-0665-z

Cites

83 of 83 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 65 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Functional connectivity exists between the cerebellum and the medial prefrontal cortex (mPFC) in mice.

    “we demonstrated functional connectivity between the cerebellum and the medial prefrontal cortex (mPFC) in mice”
    Quote in the paperSubstantial claim
  • Findingown data

    The mPFC mediates cerebellum-regulated social and repetitive/inflexible behaviors.

    “showed that the mPFC mediates cerebellum-regulated social and repetitive/inflexible behaviors”
    Quote in the paperSubstantial claim
  • Findingown data

    A circuit runs from cerebellar cortical areas Rcrus1 and posterior vermis through the cerebellar nuclei and ventromedial thalamus and culminates in the mPFC.

    “We delineated a circuit from cerebellar cortical areas Right crus 1 (Rcrus1) and posterior vermis through the cerebellar nuclei and ventromedial thalamus and culminating in the mPFC.”
    Quote in the paperSubstantial claim
  • Findingown data

    Disruptions in connectivity between the cerebellum and mPFC occur in multiple mouse models of ASD-linked genes and in individuals with ASD.

    “showed disruptions in connectivity between these regions in multiple mouse models of ASD-linked genes and in individuals with ASD”
    Quote in the paperSubstantial claim
  • Findingown data

    Activation of Purkinje cells in Rcrus1 improves social preference impairments in male PC-Tsc1 mutant mice.

    “activation of Purkinje cells (PCs) in Rcrus1 and posterior vermis improved social preference impairments and repetitive/inflexible behaviors, respectively, in male PC-Tsc1 mutant mice.”
    Quote in the paperSubstantial claim
  • May combine two claims. 36
  • Classified as probably not a substantial claim. 32
  • The quote only partly states this claim. 2

  • May combine two claims.

    Findingown data

    Motor learning, locomotor and anxiety behaviors are unaffected in PC-Tsc1 mutants and not altered by mPFC chemogenetic inhibition.

    “Motor learning, locomotor and anxiety behaviors were unaffected in PC-Tsc1 mutants and not altered by mPFC chemogenetic inhibition”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Inhibition of Rcrus1 PCs does not alter unit firing frequency in the motor cortex or in the right PRL.

    “without altering unit firing frequency in the motor cortex or in the right PRL”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    CNO-injected Gi/GFP mice (Rcrus1 PC inhibition) demonstrate impairments in social preference, social novelty, and social odor preference.

    “CNO-injected Gi/GFP mice demonstrated impairments in these behaviors as well as in social odor preference”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    VEH-injected Gi/Gi and Gi/GFP mice exhibit significant social preference and social novelty preference.

    “VEH-injected Gi/Gi and Gi/GFP mice exhibited significant social preference and social novelty preference, consistent with their control background.”
    Quote in the paperSubstantial claim

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

    Findingcitation

    Downstream thalamic and cortical areas identified are consistent with previously published results.

    “Downstream thalamic and cortical areas identified are consistent with previously published results”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    GFP-control and light-OFF Arch-expressing PC-Tsc1 mutants show social impairments.

    “GFP-control and light-OFF Arch-expressing PC-Tsc1 mutants showed social impairments consistent with their mutant backgrounds.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Photo-inhibition did not induce any change in locomotion, motor learning or anxiety.

    “The photo-inhibition did not induce any change in locomotion, motor learning or anxiety”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Laser stimulation did not affect locomotor behavior, motor learning or anxiety measures.

    “We did not observe any effects of laser stimulation on locomotor behavior, motor learning or anxiety measures that would affect the interpretation of the social or grooming behaviors”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    These cells have terminals in the left PRL mPFC in layer 1, the same area receiving input from the LN.

    “These cells also have terminals in the left PRL mPFC in the same area (layer 1) receiving input from the LN”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Inhibitory PC output emerges from the cerebellum via cerebellar nuclei.

  • May combine two claims.

    Findingown data

    Stimulation of posterior vermis PCs has no significant effect on locomotion, motor learning or anxiety.

    “Stimulation of posterior vermis PCs had no significant effect on locomotion, motor learning or anxiety”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    Chemogenetic stimulation of the posterior vermis improves behavioral flexibility in the water Y-maze in PC-Tsc1 mutant mice.

    “and improved behavioral flexibility in the water Y-maze”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Cerebellar PC dysfunction and targeted inhibition of Rcrus1 or the posterior vermis result in increased mPFC activity.

    “In this study, we showed that cerebellar PC dysfunction and targeted inhibition of Rcrus1 or the posterior vermis resulted in increased mPFC activity”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    Group MVPA yielded significant clusters in the medial prefrontal cortex and Rcrus 1 and Rcrus2 of the cerebellum.

    “Voxel-to-voxel functional connectivity maps thresholded at P < 0.001, FDR cluster corrected P < 0.05 yielded significant clusters in the medial prefrontal cortex (peak coordinates (x, y, z): 4, 38, −24; k = 260) and Rcrus 1 and Rcrus2 of the cerebellum (peak coordinates (x, y, z): 34, −82, −40; k = 18).”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Findingown data

    Thirty models with P < 0.05 in both the left mPFC and the cerebellar regions were identified.

    “Thirty models with P < 0.05 in both the left mPFC and the cerebellar regions were identified, and structural covariance analysis on these models (n = 559) and controls (n = 1,101) was also performed”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    PC-specific deletion of Tsc1 is sufficient to generate ASD-relevant social impairment and repetitive/inflexible behaviors in mice.

    “PC-specific deletion of Tsc1 is sufficient to generate ASD-relevant social impairment and repetitive/inflexible behaviors in mice5.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [5] Tsai, P. T. et al. Autistic-like behaviour and cerebellar dysfunction in Purkinje cell Tsc1 mutant mice. Nature 488, 647–651 (2012). source

      Not checked in the free run.

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

    Premisecitation

    Many studies have detailed volumetric changes in the posterior cerebellar vermis and in Rcrus1 in children with ASD.

    “Many studies have detailed volumetric changes in the posterior cerebellar vermis and in Rcrus1 in children with ASD”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [6] Pierce, K. & Courchesne, E. Evidence for a cerebellar role in reduced exploration and stereotyped behavior in autism. Biol. Psychiatry 49, 655–664 (2001). source

      Not checked in the free run.

    • cites [7] D’Mello, A. M., Crocetti, D., Mostofsky, S. H. & Stoodley, C. J. Cerebellar gray matter and lobular volumes correlate with core autism symptoms. Neuroimage Clin. 7, 631–639 (2015). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Inhibition of Rcrus1 PCs generates both social impairments and repetitive behaviors in PC-Tsc1 mutant mice.

    “We recently demonstrated that inhibition of Rcrus1 PCs could generate both social impairments and repetitive behaviors”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [8] Stoodley, C. J. et al. Altered cerebellar connectivity in autism and cerebellar-mediated rescue of autism-related behaviors in mice. Nat. Neurosci. 20, 1744–1751 (2017). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    Decreasing mPFC excitatory activity can rescue social deficits in an ASD mouse model.

    “earlier findings that decreasing mPFC excitatory activity can rescue social deficits in an ASD mouse model24”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [24] Selimbeyoglu, A. et al. Modulation of prefrontal cortex excitation/inhibition balance rescues social behavior in CNTNAP2-deficient mice. Sci. Transl. Med. 9, eaah6733 (2017). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Inhibition of mPFC is sufficient to enhance social behavior and reduce repetitive grooming in PC-Tsc1 mutants.

    “Taken together, these data demonstrate that inhibition of mPFC was sufficient to enhance social behavior and reduce repetitive grooming in PC-Tsc1 mutants.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    Sex and interaction with sex were included as covariates in the structural covariance model.

    “Sex and interaction with sex were included as covariates in the structural covariance model.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Disruptions in connectivity between the mPFC and Rcrus1 exist both in mouse models and in humans with ASD.

    “Together, these data point to disruptions in connectivity between the mPFC and Rcrus1 both in mouse models and in humans with ASD.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    A regression model tested covariates for scanner, age at scan and sex.

    “A regression model tested covariates for scanner, age at scan and sex.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Reduction of mPFC activity is sufficient to rescue both social deficits and repetitive/inflexible behaviors induced by Rcrus1 PC inhibition.

    “Taken together, these data indicate that reduction of mPFC activity is sufficient to rescue both social deficits and repetitive/inflexible behaviors induced by Rcrus1 PC inhibition”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Modulation of Rcrus1 or LN rescues only social preference behaviors but does not affect grooming.

    “In contrast, modulation of Rcrus1 or LN rescued only the social preference behaviors but did not affect grooming.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Inhibition of the PRL mPFC itself or its inputs from the VM thalamus reduces repetitive grooming and increases social preference in PC-Tsc1 mutant mice.

    “We have demonstrated that inhibition of the PRL mPFC itself or its inputs from the VM thalamus both reduces repetitive grooming and increases social preference in PC-Tsc1 mutant mice.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Photo-activation of VM terminals in the left PRL mPFC is sufficient to induce social deficits and repetitive grooming.

    “Taken together, photo-activation of VM terminals in the left PRL mPFC was sufficient to induce social deficits and repetitive grooming.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Inhibition of VM thalamus-PRL mPFC projections is sufficient to rescue both social impairments and repetitive grooming in PC-Tsc1 mutant mice.

    “These results demonstrate that inhibition of VM thalamus–PRL mPFC projections was sufficient to rescue both social impairments and repetitive grooming in PC-Tsc1 mutant mice.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    Cerebellar outputs from Rcrus1 and posterior vermis converge on VM thalamus-mPFC projections to regulate social preference and behavioral flexibility, respectively.

    “these data suggest that cerebellar outputs from Rcrus1 and posterior vermis converge on VM thalamus–mPFC projections to regulate social preference and behavioral flexibility, respectively”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Inhibitory PC output emerges from the cerebellum via cerebellar nuclei.

  • May combine two claims.

    Premisecitation

    The cerebellum provides an important source of inhibitory tone, as its output emerges solely via inhibitory PCs.

    “The cerebellum provides an important source of inhibitory tone, as its output emerges solely via inhibitory PCs41.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [41] Mouginot, D. & Gahwiler, B. H. Characterization of synaptic connections between cortex and deep nuclei of the rat cerebellum in vitro. Neuroscience 64, 699–712 (1995). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Elevated mPFC activity results in ASD-related phenotypes, and decreasing mPFC activity is sufficient to rescue abnormal behaviors in ASD models.

    “Neural circuit manipulation in combination with neural recordings provide direct evidence that elevated mPFC activity results in ASD-related phenotypes, whereas decreasing mPFC activity is sufficient to rescue abnormal behaviors in ASD models22,23,24.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [22] Kobayashi, M., Hayashi, Y., Fujimoto, Y. & Matsuoka, I. Decreased parvalbumin and somatostatin neurons in medial prefrontal cortex in BRINP1-KO mice. Neurosci. Lett. 683, 82–88 (2018). source

      Not checked in the free run.

    • cites [23] Yizhar, O. et al. Neocortical excitation/inhibition balance in information processing and social dysfunction. Nature 477, 171–178 (2011). source

      Not checked in the free run.

    • cites [24] Selimbeyoglu, A. et al. Modulation of prefrontal cortex excitation/inhibition balance rescues social behavior in CNTNAP2-deficient mice. Sci. Transl. Med. 9, eaah6733 (2017). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    E/I imbalance contributes to ASD.

    “One prominent hypothesis is that excitation/inhibition (E/I) imbalance contributes to ASD37.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [37] Rubenstein, J. L. & Merzenich, M. M. Model of autism: increased ratio of excitation/inhibition in key neural systems. Genes Brain Behav. 2, 255–267 (2003). source

      Not checked in the free run.

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

    Premisecitation

    Synaptic abnormalities result in neuronal hyperactivity and altered brain connectivity.

    “synaptic abnormalities resulting in neuronal hyperactivity and altered brain connectivity39”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [39] Contractor, A., Klyachko, V. A. & Portera-Cailliau, C. Altered neuronal and circuit excitability in fragile X syndrome. Neuron 87, 699–715 (2015). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    The posterior cerebellar vermis regulates social behaviors and behavioral flexibility.

    “These results support our hypothesis that the posterior cerebellar vermis regulates social behaviors and behavioral flexibility.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Distinct cerebellar regions regulate specific behaviors, with Rcrus1 stimulation sufficient to ameliorate social behaviors and posterior vermis stimulation sufficient to improve repetitive/inflexible behaviors.

    “Our data further demonstrate that distinct cerebellar regions regulate specific behaviors, with Rcrus1 stimulation sufficient to ameliorate social behaviors, whereas posterior vermis stimulation is sufficient to improve repetitive/inflexible behaviors.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    Disruption in the delineated cerebellum-mPFC circuits would be predicted to disrupt the ability to generate predictions, validate timing and update parameters, with a detrimental effect on social interaction.

    “from these models, disruption in these delineated circuits that connect the cerebellum and mPFC would be predicted to result in disruption in the ability to generate predictions, validate timing and update those parameters, which, all told, would be expected to have a detrimental effect on social interaction.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Precise targeting of human mPFC circuits presents a therapeutic challenge owing to the dense processing occurring in the mPFC and because the mPFC offers a less accessible target for noninvasive neuromodulation.

    “However, precise targeting of human mPFC circuits presents a therapeutic challenge owing to the dense processing occurring in the mPFC and because the mPFC offers a less accessible target for noninvasive neuromodulation46.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [46] Carmi, L. et al. Clinical and electrophysiological outcomes of deep TMS over the medial prefrontal and anterior cingulate cortices in OCD patients. Brain Stimulation 11, 158–165 (2018). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    These models involve closed-loop interactions between the cortex and cerebellum.

    “These models involve closed-loop interactions between the cortex and cerebellum19.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [19] Buckner, R. L., Krienen, F. M., Castellanos, A., Diaz, J. C. & Yeo, B. T. The organization of the human cerebellum estimated by intrinsic functional connectivity. J. Neurophysiol. 106, 2322–2345 (2011). source

      Not checked in the free run.

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

    Premisecitation

    The mPFC has key roles in socio-emotional processing and critical roles in executive control of behavior.

    “the mPFC has been demonstrated to have key roles in socio-emotional processing44 and to have critical roles in executive control of behavior.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [44] Bicks, L. K., Koike, H., Akbarian, S. & Morishita, H. Prefrontal cortex and social cognition in mouse and man. Front. Psychol. 6, 1805 (2015). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    The cerebellum is a more accessible anatomic area and might offer similar, if not greater, therapeutic value for neuropsychiatric disorders.

    “Alternatively, the cerebellum, a more accessible anatomic area, might offer similar, if not greater, therapeutic value for neuropsychiatric disorders47.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [47] van Dun, K., Bodranghien, F., Manto, M. & Marien, P. Targeting the cerebellum by noninvasive neurostimulation: a review. Cerebellum 16, 695–741 (2017). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Cerebellar output is negatively affected in ASD owing to significant PC loss and decreased cerebellar activity.

    “This output is negatively affected in ASD owing to significant PC loss and decreased cerebellar activity2,5,42.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [2] Fatemi, S. H. et al. Consensus paper: pathological role of the cerebellum in autism. Cerebellum 11, 777–807 (2012). source

      Not checked in the free run.

    • cites [5] Tsai, P. T. et al. Autistic-like behaviour and cerebellar dysfunction in Purkinje cell Tsc1 mutant mice. Nature 488, 647–651 (2012). source

      Not checked in the free run.

    • cites [42] Ryu, Y. H. et al. Perfusion impairments in infantile autism on technetium-99m ethyl cysteinate dimer brain single-photon emission tomography: comparison with findings on magnetic resonance imaging. Eur. J. Nucl. Med. 26, 253–259 (1999). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Interpretationown data

    The cerebellum plays roles in the modulation and execution of complex behaviors.

    “Our results further understanding of the roles that the cerebellum plays in the modulation and execution of complex behaviors”
    Quote in the paperSubstantial claim

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

    Interpretationcitation

    Additional cortical and sub-cortical regions might also be regulated by the cerebellum to contribute to ASD-relevant behaviors.

    “it is likely that additional cortical, including the clinically implicated right-sided prefrontal cortices20, and other sub-cortical regions might also be regulated by the cerebellum to contribute to ASD-relevant behaviors19.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [20] Gilbert, S. J., Bird, G., Brindley, R., Frith, C. D. & Burgess, P. W. Atypical recruitment of medial prefrontal cortex in autism spectrum disorders: an fMRI study of two executive function tasks. Neuropsychologia 46, 2281–2291 (2008). source

      Not checked in the free run.

    • cites [19] Buckner, R. L., Krienen, F. M., Castellanos, A., Diaz, J. C. & Yeo, B. T. The organization of the human cerebellum estimated by intrinsic functional connectivity. J. Neurophysiol. 106, 2322–2345 (2011). source

      Not checked in the free run.

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

    Study design and conductown data

    Pyramidal neurons were distinguished from interneurons by their shape, spiking pattern and action potential, and their identity was confirmed by their large apical dendrites visualized with Neurobiotin staining.

    “Pyramidal neurons were distinguished from interneurons by their shape, spiking pattern and action potential. In addition, their identity was confirmed by their large apical dendrites, which were visualized by using Neurobiotin staining.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Stimulation of the cerebellum has been shown to positively alter behaviors while affecting downstream prefrontal cortex function in preclinical studies of schizophrenia.

    “In fact, stimulation of the cerebellum has been shown to positively alter behaviors while affecting downstream prefrontal cortex function in preclinical studies of schizophrenia48,49.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [48] Parker, K. L. et al. Delta-frequency stimulation of cerebellar projections can compensate for schizophrenia-related medial frontal dysfunction. Mol. Psychiatry 22, 647–655 (2017). source

      Not checked in the free run.

    • cites [49] Brady, R. O., Jr. et al. Cerebellar-prefrontal network connectivity and negative symptoms in schizophrenia. Am. J. Psychiatry 176, 512–520 (2019). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Littermate controls were used for all behavioral experiments.

    “Littermate controls were used for all behavioral experiments.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    Mice were randomly assigned to groups for viral injection type.

    “Mice were randomly assigned to groups for viral injection type.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Mice were randomly assigned to treatment groups.

    “Mice were randomly assigned to treatment groups.”
    Quote in the paperNot a substantial claim

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

    Study design and conductreasoning

    Experimenters were not blinded to genotype for MRI scanning, registration and volume measurements because the process was automated.

    “Given that the MRI scanning, registration and volume measurements were an automated process, the experimenters were not blinded to genotype.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductown data

    After quality control, 902 scans (425 ASD and 477 control) from 648 individuals (285 ASD and 368 control) were considered for analysis.

    “After quality control, 902 scans (425 ASD and 477 control) from 648 individuals (285 ASD and 368 control) were considered for analysis.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    P values were corrected for multiple comparisons using the Benjamini-Hochberg procedure.

    “P values were corrected for multiple comparisons using the Benjamini–Hochberg procedure; these FDR-corrected P values are herein referred to as q values.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Starting treatment was randomly assigned and then alternated between VEH and CNO treatments.

    “Starting treatment was randomly assigned and then alternated between VEH and CNO treatments.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    Recordings were included only when a GΩ seal formed before whole-cell access.

    “Recordings were included only when a GΩ seal formed before whole-cell access.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    The results were largely identical when analyses were repeated using log regressions.

    “To ensure that our brain volume covarying strategy did not unduly influence the results, we repeated the analyses using log regressions as in ref. 75. The results were largely identical.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [75] Reardon, P. K. et al. Normative brain size variation and brain shape diversity in humans. Science 360, 1222–1227 (2018). source

      Not checked in the free run.

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

    Premisecitation

    There is a sensitive period for social behavior that extends into adulthood, whereas the sensitive period for repetitive behaviors closes before adulthood.

    “Our previous findings have suggested that there is a sensitive period for social behavior that extends into adulthood, whereas the sensitive period for repetitive behaviors closes before adulthood50.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [50] Tsai, P. T. et al. Sensitive periods for cerebellar-mediated autistic-like behaviors. Cell Rep. 25, 357–367 (2018). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Recording locations were verified post hoc via electrolytic lesions and histologic analysis of 50-μm coronal sections.

    “Recording locations were verified post hoc via electrolytic lesions and histologic analysis of 50-μm coronal sections.”
    Quote in the paperSubstantial claim

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

    Study design and conductcitation

    The sample size estimate was performed based on a repeated-measures ANOVA design (mixed-model ANOVA with covariance structure of compound symmetry) using PASS 12.

    “The sample size estimate was performed based on a repeated-measures ANOVA design (mixed-model ANOVA with covariance structure of compound symmetry) using PASS 12 to estimate sample size5,8.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [5] Tsai, P. T. et al. Autistic-like behaviour and cerebellar dysfunction in Purkinje cell Tsc1 mutant mice. Nature 488, 647–651 (2012). source

      Not checked in the free run.

    • cites [8] Stoodley, C. J. et al. Altered cerebellar connectivity in autism and cerebellar-mediated rescue of autism-related behaviors in mice. Nat. Neurosci. 20, 1744–1751 (2017). source

      Not checked in the free run.

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

    Study design and conductnone given

    Repeated testing in a specific behavioral paradigm was separated by 1 week to minimize previous exposure to the task.

    “To minimize previous exposure to the task, repeated testing in a specific behavioral paradigm was separated by 1 week.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    All behavioral assays were performed by examiners blinded to genotype.

    “All behavioral assays were performed by examiners blinded to genotype.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    Outlier tests were conducted using the robust regression outlier test in the Prism software suite to determine if data should be excluded.

    “Outlier tests were also conducted using the robust regression outlier test in the Prism software suite to determine if data should be excluded.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Electrophysiological data were tested for normality of distribution with a one-tailed Kolmogorov-Smirnov test.

    “Electrophysiological data were analyzed with Prism software and tested for normality of distribution with a one-tailed Kolmogorov–Smirnov test.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    Six data points were found to be outliers across all mini EPSC/IPSC recordings and were excluded.

    “Six datum were found to be outliers across all mini EPSC/IPSC recordings and were excluded.”
    Quote in the paperNot a substantial claim

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

    Study design and conductreasoning

    For rotarod and water maze testing, mice were randomly assigned to one treatment group and no repeat testing was performed.

    “For rotarod and water maze testing, which were performed in the weeks after the above testing, mice were randomly assigned to one treatment group (either VEH or CNO), and no repeat testing was performed owing to the learning component involved in these tests.”
    Quote in the paperNot a substantial claim

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

    Study design and conductcitation

    Chemogenetic studies involved controls: GFP infection (VEH and CNO) and DREADD infection (VEH and CNO) for all behavioral studies to control for DREADD activation and potential effects of VEH and to control for CNO off-target effects.

    “Chemogenetic studies involved the following controls: GFP infection (VEH and CNO) and designer receptors exclusively activated by designer drugs (DREADD) infection (VEH and CNO) for all behavioral studies to control for DREADD activation and potential effects of VEH and to control for CNO off-target effects76.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [76] Roth, B. L. DREADDs for neuroscientists. Neuron 89, 683–694 (2016). source

      Not checked in the free run.

63 of 63

  • The paper relies on this without support.

    The circuits connecting the cerebellum to other brain areas to orchestrate complex behaviors such as those altered in ASDs remain unknown.

    “Additionally, the circuits connecting the cerebellum to other brain areas to orchestrate complex behaviors such as those altered in ASDs remain unknown.”

  • The paper relies on this without support.

    Inhibitory PC output emerges from the cerebellum via cerebellar nuclei.

    “Because inhibitory PC output emerges from the cerebellum via cerebellar nuclei (CN), we identified the CN receiving Rcrus1 PC output.”

  • The paper relies on this without support.

    Cerebellar-prefrontal circuits and their regulation of ASD-relevant behaviors are conserved between mice and humans.

    “Our findings provide critical insights into how the cerebellum regulates ASD-relevant behaviors, while also offering important preclinical evidence to support a potential cerebellar therapeutic target of neuromodulation through which the therapeutic benefits of mPFC modulation could be leveraged to benefit ASD behaviors.”

  • The authors state this as an assumption.

    For all other data, the data distribution was assumed to be normal, but this was not formally tested.

    “For all other data, data distribution was assumed to be normal, but this was not formally tested.”
4 of 4

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Previous reports implicating the cerebellum in the pathogenesis of social impairments in the Tsc1 KO model of ASD do not generalize across ASD.

    “these results suggest that previous reports implicating the cerebellum in the pathogenesis of social impairments in the Tsc1 KO model of ASD (Kelly et al., 2020; Tsai et al., 2012) do not generalize across ASD”

    From Parallel social information processing circuits are differentially impacted in autism (2020) · cites this paper as Kelly et al., 2020

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.

  • The same reversal-learning comparison names two different mazes

    Minor

    The results describe the Gi/GFP group's reversal task as a water T-maze but the Gi/Gi group's reversal task as a water Y-maze, while citing the same figure panels for both.

  • Electrophysiology animal sex is reported inconsistently

    Minor

    The mice methods say only males were used for electrophysiological recordings, but the in vivo electrophysiology methods say both males and females were used.

2 of 2

Cites

Flagged first, then by citations

Research works printed but not linked: 2

  1. Paxinos, G. & Franklin, K. Paxinos and Franklin’s the Mouse Brain in Stereotaxic Coordinates 4th edn (Academic Press, 2012).
  2. Watson, C., Paxinos, G. & Puelles, L. The Mouse Nervous System (Academic Press, 2011).

Cited by

Sorted by citations

Related

Authors
Kelly, E., Meng, F., Fujita, H., Morgado, F., Kazemi, Y., Rice, L. C., Ren, C., Escamilla, C. O., Gibson, J. M., Sajadi, S., Pendry, R. J., Tan, T., Ellegood, J., Basson, M. A., Blakely, R. D., Dindot, S. V., Golzio, C., Hahn, M. K., Katsanis, N., Robins, D. M., Silverman, J. L., Singh, K. K., Wevrick, R., Taylor, M. J., Hammill, C., Anagnostou, E., Pfeiffer, B. E., Stoodley, C. J., Lerch, J. P., du Lac, S., & Tsai, P. T.
Affiliations
  • The University of Texas Southwestern Medical Center (US)
  • Johns Hopkins University (US)
  • University of Toronto (CA)
  • American University (US)
  • Hospital for Sick Children (CA)
  • King's College London (GB)
  • Florida Atlantic University (US)
  • Texas A&M University (US)
  • Centre National de la Recherche Scientifique (FR)
  • Northwestern University (US)
  • University of Michigan (US)
  • University of California, Davis (US)
  • McMaster University (CA)
  • University of Alberta (CA)
Corresponding author
Tsai, Peter T.
ORCID
Venue
Nature Neuroscience
Publisher
Springer Science and Business Media LLC
Published
2020-07-13
Volume
23
Issue
9
Pages
1102-1110
ISSN
  • 1097-6256 (ISSN-L)
  • 1546-1726
DOI
10.1038/s41593-020-0665-z
PMC
PMC7483861
OpenAlex
W3041250095
Semantic Scholar
88c1b417a376de87989030480ac1dacd732efa1e
Cited by
280
References
83
Citations by year
2026: 31 · 2025: 45 · 2024: 59 · 2023: 55 · 2022: 46 · 2021: 39 · 2020: 5
FWCI
19.47
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Neuroscience (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • HAL (submittedVersion, cc-by-nc)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
  • U.S. Department of Health & Human Services | NIH | National Institute of Neurological Disorders and Stroke (NS107004, NS095232, NS105039, NS083733)
  • Autism Speaks
  • Gouvernement du Canada | Instituts de Recherche en Santé du Canada | CIHR Skin Research Training Centre
  • Ontario Brain Institute
  • U.S. Department of Health & Human Services | NIH | National Institute of Mental Health (MH106957, MH094268, MH116882)
  • Tuberous Sclerosis Alliance
Topics
  • Life Sciences › Neuroscience › Cognitive Neuroscience › Autism Spectrum Disorder Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetics and Neurodevelopmental Disorders
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
Keywords
autism spectrum disorder, cerebellum, medial prefrontal cortex, social behavior, repetitive behavior, Purkinje cells, posterior vermis, ventromedial thalamus, cerebellar nuclei
MeSH terms
Autism Spectrum Disorder, Animals, Cerebellum, Male, Mice, Mutant Strains, Neural Pathways, Prefrontal Cortex, Mice

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 2026-10-09.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-09.

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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