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Cited by 105Genetics and Neurodevelopmental DisordersAutism Spectrum Disorder ResearchNeuroscience and Neuropharmacology Research

Autism-like social deficit generated by Dock4 deficiency is rescued by restoration of Rac1 activity and NMDA receptor function.

Open access
Guo, Daji, Peng, Yinghui, Wang, Laijian, Sun, Xiaoyu, Wang, Xiaojun, Liang, Chunmei, Yang, Xiaoman, Li, Shengnan +4 more·2021·Molecular Psychiatry
DOI 10.1038/s41380-019-0472-7PMC PMC8159750
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Abstract

Genetic studies of autism spectrum disorder (ASD) have revealed multigene variations that converge on synaptic dysfunction. DOCK4, a gene at 7q31.1 that encodes the Rac1 guanine nucleotide exchange factor Dock4, has been identified as a risk gene for ASD and other neuropsychiatric disorders. However, whether and how Dock4 disruption leads to ASD features through a synaptic mechanism remain unexplored. We generated and characterized a line of Dock4 knockout (KO) mice, which intriguingly displayed a series of ASD-like behaviors, including impaired social novelty preference, abnormal isolation-induced pup vocalizations, elevated anxiety, and perturbed object and spatial learning. Mice with conditional deletion of Dock4 in hippocampal CA1 recapitulated social preference deficit in KO mice. Examination in CA1 pyramidal neurons revealed that excitatory synaptic transmission was drastically attenuated in KO mice, accompanied by decreased spine density and synaptic content of AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid)- and NMDA (N-methyl-D-aspartate)-type glutamate receptors. Moreover, Dock4 deficiency markedly reduced Rac1 activity in the hippocampus, which resulted in downregulation of global protein synthesis and diminished expression of AMPA and NMDA receptor subunits. Notably, Rac1 replenishment in the hippocampal CA1 of Dock4 KO mice restored excitatory synaptic transmission and corrected impaired social deficits in these mice, and pharmacological activation of NMDA receptors also restored social novelty preference in Dock4 KO mice. Together, our findings uncover a previously unrecognized Dock4-Rac1-dependent mechanism involved in regulating hippocampal excitatory synaptic transmission and social behavior.

AI snapshot

Written from the paper’s full text.

TL;DR

Dock4 knockout mice display autism-like social, vocalization, anxiety, and learning deficits, with reduced hippocampal excitatory synaptic transmission and AMPA/NMDA receptor content. Restoring Rac1 activity or NMDA receptor function in hippocampal CA1 rescues social novelty preference, implicating a Dock4-Rac1-dependent pathway in social behavior.

Key findings

  • Dock4 KO mice showed impaired social novelty preference, more isolation-induced pup calls at postnatal days 3, 6, 9, and 12, longer total call duration, elevated anxiety, and perturbed object/spatial learning; conditional Dock4 deletion in hippocampal CA1 recapitulated the social preference deficit.
  • In CA1 pyramidal neurons, KO mice had smaller mEPSC amplitude, unchanged mIPSCs, a substantially smaller E/I ratio, reduced AMPAR- and NMDAR-mediated EPSCs with a more prominent NMDAR reduction, a lowered NMDAR/AMPAR ratio, weaker ifenprodil blockade of NMDAR-EPSCs, and significantly reduced LTP.
  • KO hippocampus showed decreased dendritic spines in CA1 and dentate gyrus and reduced synaptic expression of GluA2, GluN1, GluN2A, and GluN2B; global expression of these subunits was still reduced by 10%–30%, while mRNA levels were unaltered.
  • Dock4 deficiency reduced active Rac1 and global protein synthesis; Rac1 overexpression restored protein synthesis and receptor subunit levels in vitro, rescued mEPSC and LTP in KO neurons, and corrected social novelty preference in vivo. D-cycloserine rapidly restored social novelty preference in KO mice, but the effect diminished by 7 days; the AMPAR positive allosteric modulator PF-4774857 had no effect.

Methods

  • Generated whole-body Dock4 KO mice via Cre-LoxP recombination and used conditional deletion by AAV-Cre delivery into hippocampal CA1 of Dock4 fl/fl mice.
  • Behavioral analyses used Dock4 KO, HET, and WT littermates of both genders aged 3–6 months, except pup-retrieval assays used virgin females; tests included three-chamber social approach/novelty, ultrasonic vocalizations, elevated zero-maze, novel object recognition, Y-maze, Morris water-maze, nesting, and pup retrieval.
  • Synaptic function was assessed by whole-cell electrophysiology in hippocampal CA1 pyramidal neurons; protein expression and synthesis were measured by Western blotting, puromycin labeling, Rac1 activity assays, lentiviral Rac1 expression, and pharmacological treatments including DCS and PF-4774857.

Limitations

  • The paper does not state a dedicated limitations section.
  • Evident from the text: findings are based on mouse models and cultured neurons, with no human tissue or human behavioral validation reported.
  • Evident from the text: some phenotypes were sex-specific or based on small subgroups; a small population of female HET (1.7%) and KO (9.1%) mice showed stereotyped behavior and was excluded from subsequent studies, and DCS rescue of social novelty preference diminished by 7 days after treatment.

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

Integrity signals

Registry

Crossref · 10.1038/s41380-019-0472-7

Cites

66 of 66 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

35 of 59 cited sources checked · 18 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    Restoration of normal Rac1 activity or its downstream effectors corrects autism-like behaviors and synaptic abnormalities.

    “Supporting this notion, restoration of normal activity of Rac1 or its downstream effectors was demonstrated as a successful approach for correcting autism-like behaviors and synaptic abnormalities 12–15.”
    Quote in the paperSubstantial claim2 of 4 sources checked
    • cites [12] Dolan BM, Duron SG, Campbell DA, Vollrath B, Shankaranarayana Rao BS, Ko HY, et al. Rescue of fragile X syndrome phenotypes in Fmr1 KO mice by the small-molecule PAK inhibitor FRAX486. Proc Natl Acad Sci USA. 2013;110:5671–6. source

      The Discussion reports that the PAK inhibitor FRAX486 reversed abnormal spine density and ameliorated autism-like behaviors in Fmr1 KO mice; the paper identifies PAK as downstream of Rac1. It does not demonstrate restoration of normal Rac1 activity, or establish that PAK activity itself was restored to a normal level. The demonstrated correction is limited to the mouse model studied. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [13] Duffney LJ, Zhong P, Wei J, Matas E, Cheng J, Qin L, et al. Autism-like deficits in shank3-deficient mice are rescued by targeting actin regulators. Cell Rep. 2015;11:1400–13. source
    • cites [14] Li J, Chai A, Wang L, Ma Y, Wu Z, Yu H, et al. Synaptic P-Rex1 signaling regulates hippocampal long-term depression and autism-like social behavior. Proc Natl Acad Sci USA. 2015;112:E6964–72. source
    • cites [15] Martinez LA, Tejada-Simon MV. Pharmacological rescue of hippocampal fear learning deficits in fragile X syndrome. Mol Neurobiol. 2017;55:5951–61. source
  • Premisecitation

    DOCK4 is a risk gene for ASD and other neuropsychiatric disorders.

    “Dock4, a member of the Dock (dedicator of cytokinesis) family, is an atypical Rac1 GEF linked to ASD and other neuropsychiatric disorders 17.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [17] Shi L. Dock protein family in brain development and neurological disease. Commun Integr Biol. 2013;6:e26839. source
  • Premisecitation

    Rac1 is a converging factor downstream of numerous proteins encoded by high-risk ASD genes.

    “Rac1 has been identified as a converging factor downstream of numerous proteins encoded by high-risk ASD genes, such as SHANK 4–6, FMR1 (fragile X mental retardation-1) 7, 8, NRXN 9, and AUTS2 10.”
    Quote in the paperSubstantial claim4 of 6 sources checked
    • cites [4] Duffney LJ, Wei J, Cheng J, Liu W, Smith KR, Kittler JT, et al. Shank3 deficiency induces NMDA receptor hypofunction via an actin-dependent mechanism. J Neurosci. 2013;33:15767–78. source
    • cites [5] Sarowar T, Grabrucker AM. Actin-dependent alterations of dendritic spine morphology in shankopathies. Neural Plast. 2016;2016:8051861. source

      Section 5 describes SHANK3 interactions that regulate Rac1 and reports reduced Rac1 activity in Shank3-knockout mice, supporting the SHANK example. But section 4 calls FMRP an indirect downstream effector of Rac1—the reverse of the claimed direction—and the cited passages do not establish Rac1 as downstream of NRXN or AUTS2. The paper therefore supports part of the proposed convergence, not the full claim. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [6] Monteiro P, Feng G. SHANK proteins: roles at the synapse and in autism spectrum disorder. Nat Rev Neurosci. 2017;18:147–57. source
    • cites [7] Pyronneau A, He Q, Hwang JY, Porch M, Contractor A, Zukin RS. Aberrant Rac1-cofilin signaling mediates defects in dendritic spines, synaptic function, and sensory perception in fragile X syndrome. Sci Signal. 2017;10:eaan0852. source
    • cites [8] Martinez LA, Tejada-Simon MV. Increased training intensity induces proper membrane localization of actin remodeling proteins in the hippocampus preventing cognitive deficits: implications for fragile X syndrome. Mol Neurobiol. 2017;55:4529–42. source
    • cites [9] Rui M, Qian J, Liu L, Cai Y, Lv H, Han J, et al. The neuronal protein Neurexin directly interacts with the Scribble-Pix complex to stimulate F-actin assembly for synaptic vesicle clustering. J Biol Chem. 2017;292:14334–48. source
  • Premisecitation

    Several microdeletions lead to loss of various lengths of the C-terminal portion of Dock4 protein, which is responsible for its Rac1-activating ability.

    “For instance, several microdeletions lead to loss of various lengths of the C-terminal portion of Dock4 protein 19, 21, 26, which is responsible for its Rac1-activating ability 27.”
    Quote in the paperSubstantial claim2 of 4 sources checked
    • cites [19] Pagnamenta AT, Bacchelli E, de Jonge MV, Mirza G, Scerri TS, Minopoli F, et al. Characterization of a family with rare deletions in CNTNAP5 and DOCK4 suggests novel risk loci for autism and dyslexia. Biol Psychiatry. 2010;68:320–8. source

      The Results report distinct DOCK4 deletions: a 594-kb IMMP2L–DOCK4 deletion and a 132-kb deletion of DOCK4 exons 38–52. They do not establish that these deletions produce proteins missing different lengths of the C-terminal portion; for the 594-kb deletion, the reported fusion transcript is likely subject to nonsense-mediated decay. The Rac1-activation clause has a separate citation in the citing sentence, although this paper’s Discussion does describe DOCK4 as a Rac1-activating factor. Page numbers were not available in our copy. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [21] Iossifov I, O’Roak BJ, Sanders SJ, Ronemus M, Krumm N, Levy D, et al. The contribution of de novo coding mutations to autism spectrum disorder. Nature. 2014;515:216–21. source

      Citation 26 is attached to the claim about microdeletions removing portions of Dock4’s C-terminus; the Rac1-activating function is separately cited to reference 27. The supplied Results and Discussion passages discuss de novo coding mutations generally, but neither they nor searches for “Dock4” and “DOCK4 microdeletions C-terminal portion” yielded a passage addressing those deletions. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [26] Kushima I, Aleksic B, Nakatochi M, Shimamura T, Okada T, Uno Y, et al. Comparative analyses of copy-number variation in autism spectrum disorder and schizophrenia reveal etiological overlap and biological insights. Cell Rep. 2018;24:2838–56. source
    • cites [27] Xiao Y, Peng Y, Wan J, Tang G, Chen Y, Tang J, et al. The atypical guanine nucleotide exchange factor Dock4 regulates neurite differentiation through modulation of Rac1 GTPase and actin dynamics. J Biol Chem. 2013;288:20034–45. source
  • Premisecitation

    Recent studies have identified DOCK4 variations associated with ASD, including single nucleotide variations and chromosome microdeletions or duplications.

    “Recent studies have identified a number of DOCK4 variations associated with ASD, including both single nucleotide variations (SNVs) and chromosome microdeletions or duplications (Supplementary Table 1) 18–26.”
    Quote in the paperSubstantial claim7 of 9 sources checked
    • cites [18] Maestrini E, Pagnamenta AT, Lamb JA, Bacchelli E, Sykes NH, Sousa I, et al. High-density SNP association study and copy number variation analysis of the AUTS1 and AUTS5 loci implicate the IMMP2L-DOCK4 gene region in autism susceptibility. Mol Psychiatry. 2010;15:954–68. source

      The Discussion reports a replicated association for the DOCK4 SNP rs2217262 and suggests that a DOCK4 deletion may influence ASD susceptibility. The Results describe a duplication involving DOCK4, but the Discussion says duplications may not be damaging; this paper does not establish a duplication associated with ASD. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [19] Pagnamenta AT, Bacchelli E, de Jonge MV, Mirza G, Scerri TS, Minopoli F, et al. Characterization of a family with rare deletions in CNTNAP5 and DOCK4 suggests novel risk loci for autism and dyslexia. Biol Psychiatry. 2010;68:320–8. source
    • cites [20] Liang S, Wang XL, Zou MY, Wang H, Zhou X, Sun CH, et al. Family-based association study of ZNF533, DOCK4 and IMMP2L gene polymorphisms linked to autism in a northeastern Chinese Han population. J Zhejiang Univ Sci B. 2014;15:264–71. source

      The Discussion reports an association between the DOCK4 SNP rs2217262 and autism, and describes genomic disruption of DOCK4 in relation to autism. The References identify a study of rare DOCK4 deletions, but our copy does not identify DOCK4 duplications associated with ASD. The claim’s duplication component is therefore unsupported by this source. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [21] Iossifov I, O’Roak BJ, Sanders SJ, Ronemus M, Krumm N, Levy D, et al. The contribution of de novo coding mutations to autism spectrum disorder. Nature. 2014;515:216–21. source

      The Introduction and Results discuss de novo coding variants and copy-number variation in ASD generally, but no passage identifies DOCK4 variants or links DOCK4 microdeletions or duplications to ASD. Searches for DOCK4 and for the specific variant types did not locate that gene in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [22] Toma C, Torrico B, Hervas A, Valdes-Mas R, Tristan-Noguero A, Padillo V, et al. Exome sequencing in multiplex autism families suggests a major role for heterozygous truncating mutations. Mol Psychiatry. 2014;19:784–90. source

      The Results section lists inherited copy-number variants in several genes, but not DOCK4. A search of our copy for DOCK4 and for DOCK4 deletions, duplications, and single-nucleotide variants found no passage addressing DOCK4 variation in ASD (page numbers unavailable). (LLM, conf=0.75)

      Experimental — accuracy not yet measured
    • cites [23] Warrier V, Chakrabarti B, Murphy L, Chan A, Craig I, Mallya U, et al. A pooled genome-wide association study of asperger syndrome. PloS ONE. 2015;10:e0131202. source

      The Discussion identifies an intronic DOCK4 SNP as nominally associated in the Asperger syndrome study, and the References list earlier work on DOCK4 deletions and copy-number variation in the IMMP2L–DOCK4 region. But the paper does not identify DOCK4 duplications specifically. It also reports that its SNP did not remain significant after correction or show a significant association in a larger autism-spectrum cohort. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [24] Shao S, Kong R, Zou L, Zhong R, Lou J, Zhou J, et al. The roles of genes in the neuronal migration and neurite outgrowth network in developmental dyslexia: single- and multiple-risk genetic variants. Mol Neurobiol. 2016;53:3967–75. source
    • cites [25] Lim ET, Uddin M, De Rubeis S, Chan Y, Kamumbu AS, Zhang X, et al. Rates, distribution and implications of postzygotic mosaic mutations in autism spectrum disorder. Nat Neurosci. 2017;20:1217–24. source

      The Introduction discusses copy-number variants and point mutations associated with ASD generally, but does not identify DOCK4. Searches of our copy for DOCK4 and DOCK4-associated SNVs, microdeletions, and duplications returned no passage addressing the gene-specific claim. No page numbers were supplied for these passages. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [26] Kushima I, Aleksic B, Nakatochi M, Shimamura T, Okada T, Uno Y, et al. Comparative analyses of copy-number variation in autism spectrum disorder and schizophrenia reveal etiological overlap and biological insights. Cell Rep. 2018;24:2838–56. source
  • May combine two claims. 14
  • Classified as probably not a substantial claim. 16
  • The quote only partly states this claim. 1

  • May combine two claims.

    Premisecitation

    Rac1 plays critical roles in modulating both the localization of neurotransmitter receptors and the architecture of excitatory synapses.

    “Rac1, a master actin regulator, plays critical roles in modulating both the localization of neurotransmitter receptors and the architecture of excitatory synapses 3.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [3] Tejada-Simon MV. Modulation of actin dynamics by Rac1 to target cognitive function. J Neurochem. 2015;133:767–79. source

      The Abstract describes Rac1 as an actin regulator important for dendritic spine morphogenesis, supporting its role in excitatory-synapse architecture. The section “Actin dynamics in neurons: link to intellectual disability” says glutamate receptors regulate Rac1, but does not say Rac1 controls the localization of neurotransmitter receptors; it describes Rac1 itself moving to the membrane after NMDA receptor activation. No page number is supplied for these passages. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

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

    Premisecitation

    Rac1 activity is tightly controlled by Rac1 guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs).

    “Rac1 activity is tightly controlled by Rac1 guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs), two protein families that directly activate and inhibit Rac1, respectively 16.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [16] Bai Y, Xiang X, Liang C, Shi L. Regulating Rac in the nervous system: molecular function and disease implication of Rac GEFs and GAPs. BioMed Res Int. 2015;2015:632450. source

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

    Premisecitation

    Both excessive and insufficient Rac1 activity induced by missense mutations in RAC1 cause developmental disorders with diverse phenotypes.

    “Moreover, both excessive and insufficient Rac1 activity induced by missense mutations in RAC1 itself cause developmental disorders presenting diverse phenotypes 11.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [11] Reijnders MRF, Ansor NM, Kousi M, Yue WW, Tan PL, Clarkson K, et al. RAC1 missense mutations in developmental disorders with diverse phenotypes. Am J Hum Genet. 2017;101:466–77. source

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

    Premisecitation

    ASD subjects commonly present co-occurring symptoms that include anxiety and intellectual disability.

    “Besides the core symptoms, ASD subjects commonly present co-occurring symptoms that include anxiety and intellectual disability 37, 38.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [37] Simonoff E, Pickles A, Charman T, Chandler S, Loucas T, Baird G. Psychiatric disorders in children with autism spectrum disorders: prevalence, comorbidity, and associated factors in a population-derived sample. J Am Acad Child Adolesc Psychiatry. 2008;47:921–9. source

      The Abstract and Rates of Disorder section report that anxiety disorders are common among the children with ASD studied, including social anxiety disorder in 29.2%. The Measures section reports a wide IQ range but does not establish that intellectual disability is a common co-occurring condition in this sample; it also does not classify intellectual disability as a symptom. Page numbers are not available in our copy. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [38] Eussen ML, Van Gool AR, Verheij F, De Nijs PF, Verhulst FC, Greaves-Lord K. The association of quality of social relations, symptom severity and intelligence with anxiety in children with autism spectrum disorders. Autism. 2013;17:723–35. source

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

    Premisecitation

    Dock4 plays critical roles in axon guidance, dendritic development, and dendritic spine formation, but these results were obtained from in vitro preparations.

    “Dock4 has been found to plays critical roles in axon guidance, dendritic development, and dendritic spine formation, but these results were obtained from in vitro preparations 27, 29–31.”
    Quote in the paperSubstantial claim1 of 4 sources checked
    • cites [27] Xiao Y, Peng Y, Wan J, Tang G, Chen Y, Tang J, et al. The atypical guanine nucleotide exchange factor Dock4 regulates neurite differentiation through modulation of Rac1 GTPase and actin dynamics. J Biol Chem. 2013;288:20034–45. source
    • cites [29] Ueda S, Fujimoto S, Hiramoto K, Negishi M, Katoh H. Dock4 regulates dendritic development in hippocampal neurons. J Neurosci Res. 2008;86:3052–61. source
    • cites [30] Ueda S, Negishi M, Katoh H. Rac GEF Dock4 interacts with cortactin to regulate dendritic spine formation. Mol Biol Cell. 2013;24:1602–13. source

      The Results section (‘Knockdown of Dock4 reduces dendritic spine density’; page unavailable) reports that Dock4 knockdown impaired spine formation in cultured rat hippocampal neurons, supporting the in vitro spine-formation part of the claim. The passages found do not establish roles for Dock4 in axon guidance or broader dendritic development. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [31] Makihara S, Morin S, Ferent J, Cote JF, Yam PT, Charron F. Polarized dock activity drives shh-mediated axon guidance. Dev Cell. 2018;46:410–25 e417. source

  • May combine two claims.

    Premisecitation

    DOCK4 is located at chromosome band 7q31.1, an autism-susceptible locus.

    “DOCK4 is located at chromosome band 7q31.1 18, 19, an autism-susceptible locus resided by several ASD-associated genes involved in development and language regulation.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [18] Maestrini E, Pagnamenta AT, Lamb JA, Bacchelli E, Sykes NH, Sousa I, et al. High-density SNP association study and copy number variation analysis of the AUTS1 and AUTS5 loci implicate the IMMP2L-DOCK4 gene region in autism susceptibility. Mol Psychiatry. 2010;15:954–68. source

      The Discussion identifies the IMMP2L/DOCK4 locus on chromosome 7 as a possible contributor to autism susceptibility, and the Introduction places the studied locus on chromosome 7q. Our copy does not specify that DOCK4 is at the narrower band 7q31.1, so that precise location is not verified. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [19] Pagnamenta AT, Bacchelli E, de Jonge MV, Mirza G, Scerri TS, Minopoli F, et al. Characterization of a family with rare deletions in CNTNAP5 and DOCK4 suggests novel risk loci for autism and dyslexia. Biol Psychiatry. 2010;68:320–8. source

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

    Premisecitation

    Variations in the gene encoding GluN2B have been frequently identified in neuropsychiatric disorders.

    “Variations in the gene encoding GluN2B have been frequently identified in neuropsychiatric disorders 62, 63, and studies on animal models have shown that GluN2B is particularly critical for social behaviors 61, 64, 65.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [62] Liu S, Zhou L, Yuan H, Vieira M, Sanz-Clemente A, Badger JD 2nd, et al. A rare variant identified within the GluN2B C-terminus in a patient with autism affects NMDA receptor surface expression and spine density. J Neurosci. 2017;37:4093–102. source
    • cites [63] Fedele L, Newcombe J, Topf M, Gibb A, Harvey RJ, Smart TG. Disease-associated missense mutations in GluN2B subunit alter NMDA receptor ligand binding and ion channel properties. Nat Commun. 2018;9:957. source

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

    Premisecitation

    Bidirectional changes of E/I ratio of synaptic inputs in different brain regions have been implicated in ASD pathology.

    “Bidirectional changes of E/I ratio of synaptic inputs in different brain regions have been implicated in ASD pathology 60.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [60] Gatto CL, Broadie K. Genetic controls balancing excitatory and inhibitory synaptogenesis in neurodevelopmental disorder models. Front Synaptic Neurosci. 2010;2:4. source

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

    Premisecitation

    p70S6K is a kinase essential for switching on translation initiation.

    “First, Rac1 appears to activate mTOR and its downstream target p70S6K (70-kDa ribosomal S6 kinase) 57, 58, a kinase essential for switching on translation initiation 48, 49.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [48] Bhattacharya A, Mamcarz M, Mullins C, Choudhury A, Boyle RG, Smith DG, et al. Targeting translation control with p70 S6 Kinase 1 inhibitors to reverse phenotypes in fragile X syndrome mice. Neuropsychopharmacology. 2016;41:1991–2000. source

      The Introduction says S6K1 promotes cap-dependent translation initiation by stimulating eIF4A activity, supporting its role in initiation. It does not say S6K1 is essential for switching initiation on; that necessity is stronger than the passage supports. No page number is provided in our copy. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [49] Berg JM, Lee C, Chen L, Galvan L, Cepeda C, Chen JY, et al. JAKMIP1, a novel regulator of neuronal translation, modulates synaptic function and autistic-like behaviors in mouse. Neuron. 2015;88:1173–91. source

      The Discussion (page unavailable) says that PABPC1 is required for translation initiation, but it does not make that statement about p70S6K. Searches for p70S6K, S6K, and the claimed kinase role found no passage in our copy addressing whether p70S6K is essential for switching on translation initiation. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Findingown data

    No Dock4 was detected in any brain region or other organs in KO mice, whereas reduced amounts of Dock4 were found in the corresponding HET tissues.

    “No Dock4 was detected in any brain region or other organs in KO mice, whereas reduced amounts of Dock4 were found in the corresponding HET tissues (Figure 1c, Supplementary Figure 3f).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Dock4 HET and KO mice exhibited similar growth rate, ingestion, and mating as WT mice.

    “Despite showing slightly lower natality (Supplementary Figure 4a), Dock4 HET and KO mice exhibited similar growth rate (weight gain; Supplementary Figure 4b,c), ingestion, and mating as WT mice did.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The Dock4 transcript was completely absent in the KO brain and reduced by approximately half in the HET brain.

    “The Dock4 transcript was completely absent in the KO brain and reduced by approximately half in the HET brain (Figure 1b, Supplementary Figure 3d).”
    Quote in the paperSubstantial claim

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

    Findingcitation

    Dock4 is expressed in various brain regions and is highly enriched in the hippocampus.

    “In accord with previous observations 27, Dock4 was found to be expressed in various brain regions and highly enriched in the hippocampus (Figure 1c).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [27] Xiao Y, Peng Y, Wan J, Tang G, Chen Y, Tang J, et al. The atypical guanine nucleotide exchange factor Dock4 regulates neurite differentiation through modulation of Rac1 GTPase and actin dynamics. J Biol Chem. 2013;288:20034–45. source

  • May combine two claims.

    Findingown data

    Male HET and KO mice and the majority of female HET and KO mice displayed mostly normal locomotion in the open field and did not show stereotyped behaviors.

    “The male HET and KO mice and the majority of the female HET and KO mice displayed mostly normal locomotion in the open field (Figure 1f) and did not show stereotyped behaviors (Supplementary Figure 5b,c).”
    Quote in the paperNot a substantial claim

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

    Findingown data

    The brain was slightly heavier in female KO mice than in female WT mice.

    “and the brain was slightly heavier in female KO mice than in female WT mice (Figure 1e).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In the social approach phase, WT, HET, and KO mice all performed normally, spending more time exploring the stranger conspecific than the object.

    “In the social approach phase, WT, HET, and KO mice all performed normally, spending more time exploring the stranger conspecific than the object (Figure 1i–k; Supplementary Figure 6f,g).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    A Dock4 KO mouse line was established and the loss of Dock4 expression in the mice was verified using qRT-PCR and western blotting.

    “A Dock4 KO mouse line was established, and the loss of Dock4 expression in the mice was verified using quantitative real-time PCR (qRT-PCR) and western blotting (Figure 1a, Supplementary Figures 2 and 3).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Brain architecture in Dock4 KO mice was grossly normal, with slight enlargement of the lateral ventricles.

    “Brain architecture in Dock4 KO mice was grossly normal, with slight enlargement of the lateral ventricles (Figure 1d, Supplementary Figure 4d–g)”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Reduction of some AMPA and NMDA subunits was also observed in prefrontal cortex, but not striatum and cerebellum of KO mice.

    “Reduction of some AMPA and NMDA subunits was also observed in prefrontal cortex, but not striatum and cerebellum of KO mice (Supplementary Figure 11).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In Dock4 KO mice, NMDAR-mediated synaptic transmission and plasticity were impaired.

    “Our findings further unravel that NMDAR hypofunction-induced failure of excitatory synaptic function is a main cause of social deficit in Dock4 KO mice, as NMDAR-mediated synaptic transmission and plasticity were impaired, and the NMDA agonist DCS restored normal social behavior.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Genetic studies of ASD have revealed multigene variations that converge on synaptic dysfunction.

    “ASD exhibits a strong genetic basis, and increasing evidence from inherited and de novo gene variations suggests a notable convergence on synapse pathophysiology in ASD, particularly dysfunction of excitatory synaptic transmission 1, 2.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [1] Bourgeron T. From the genetic architecture to synaptic plasticity in autism spectrum disorder. Nat Rev Neurosci. 2015;16:551–63. source
    • cites [2] Joensuu M, Lanoue V, Hotulainen P. Dendritic spine actin cytoskeleton in autism spectrum disorder. Prog neuro-Psychopharmacol Biol Psychiatry. 2018;84(Pt B):362–81. source

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

    Study design and conductnone given

    For virus injection and drug treatment studies, animals were randomly assigned to two groups according to gender and body weight.

    “For virus injection and drug treatment studies, animals were randomly assigned to two groups, each according to gender (male: female ratio is roughly 1:1) and body weight.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Dock4 KO mice exhibited both language and social deficits, and displayed increased tendency of stereotyped behavior.

    “Together, characterizations of the ASD core symptoms revealed that Dock4 KO mice exhibited both language and social deficits, and displayed increased tendency of stereotyped behavior.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Ultrasonic vocalizations in pups when isolated from their dam measure language communication in mice.
    • Rests on an assumption: The pup-retrieval assay measures maternal behavior in mice.
    • Rests on an assumption: The hyperactivity, stereotyped behavior, and social/communication deficits observed in Dock4 mutant mice are autism-like or ASD core symptoms.

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

    Study design and conductnone given

    Neuro-2a cells were routinely tested for mycoplasma contamination-free before use.

    “Neuro-2a cells were purchased from ATCC and were routinely tested for mycoplasma contamination-free before use.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Investigators were blind to genotype or treatment of the mice being tested during all experiments.

    “Investigators were blind to genotype or treatment of the mice being tested during all experiments.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Sample size chosen for each study was established by previous pilot and published studies.

    “Sample size chosen for each study was established by previous pilot and published studies.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Normal distribution and similar variance within each comparison group of data were checked before any parametric analyses.

    “normal distribution and similar variance within each comparison group of data were checked before any parametric analyses; P < 0.05 was considered significant.”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Interpretationown data

    LTD is intact in KO mice.

    “Whereas both forms of LTD were hardly induced at adult as reported 40, they are similarly induced in WT and KO hippocampus at young postnatal age, suggesting that LTD is intact in KO mice (Supplementary Figure 9).”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Both forms of LTD were hardly induced at adult as reported.

    “Whereas both forms of LTD were hardly induced at adult as reported 40, they are similarly induced in WT and KO hippocampus at young postnatal age, suggesting that LTD is intact in KO mice (Supplementary Figure 9).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [40] Khoo GH, Lin YT, Tsai TC, Hsu KS. Perineuronal nets restrict the induction of long-term depression in the mouse hippocampal CA1 region. Mol Neurobiol. 2019;56:6436–50. source
29 of 29

  • The paper relies on this without support.

    The 61-aa truncated Dock4 protein produced by the null allele is nonfunctional.

    “Because of the deletion and the resulting frameshift, the null allele generates a 61-aa truncated protein (41 aa of Dock4 N-terminal sequence and 20 aa of frame-shifted nonsense sequence) instead of the full-length 1978 aa protein (Supplementary Figure 2 and 3, Supplementary methods).”

  • The paper relies on this without support.

    Social novelty preference deficit in mice is a valid model of ASD social deficits.

    “We generated and characterized a line of Dock4 knockout (KO) mice, which intriguingly displayed a series of ASD-like behaviors, including impaired social novelty preference, abnormal isolation-induced pup vocalizations, elevated anxiety, and perturbed object and spatial learning.”

  • The paper relies on this without support.

    Many DOCK4 variations associated with ASD result in loss-of-function.

    “Many of these variations are believed to result in loss-of-function of DOCK4.”

  • The paper relies on this without support.

    Nesting ability in mice tightly correlates with social behaviors such as parenting and reproduction.

    “First, the ability of nesting, a nature in mice that tightly correlates with social behaviors such as parenting and reproduction, was evaluated.”

  • The paper relies on this without support.

    Ultrasonic vocalizations in pups when isolated from their dam measure language communication in mice.

    “To investigate language communication in Dock4 KO mice, we evaluated ultrasonic vocalizations in pups when isolated from their dam within the first two weeks after birth.”

  • The paper relies on this without support.

    The pup-retrieval assay measures maternal behavior in mice.

    “Second, maternal behavior was measured in a pup-retrieval assay by using virgin female mice.”

  • The paper relies on this without support.

    The hyperactivity, stereotyped behavior, and social/communication deficits observed in Dock4 mutant mice are autism-like or ASD core symptoms.

    “Notably, a small population of female HET (1.7%) and KO (9.1%) mice showed autism-like hyperactivity and stereotyped behavior, such as continual circling in the home cage and open field (Figure 1f, Supplementary Figure 5a, Supplementary Videos 1–3).”

  • The paper relies on this without support.

    NMDAR function is important for the induction of both long-term potentiation (LTP) and long-term depression (LTD).

    “Because NMDAR function is important for the induction of both long-term potentiation (LTP) and long-term depression (LTD), we examined whether these two forms of synaptic plasticity are altered by Dock4 deficiency.”

  • The paper relies on this without support.

    Rac1 is a well-studied actin regulator required for normal synaptic function that acts through diverse downstream effectors.

    “Rac1 is a well-studied actin regulator required for normal synaptic function that acts through diverse downstream effectors.”

  • The paper relies on this without support.

    Rac1 is a converging factor that receives signals from different ASD genes.

    “Thus, Rac1, as a converging factor that receiving signals from different ASD genes, might exert dual effects on actin dynamics as well as protein synthesis depending on the context of synaptic regulation.”

  • The paper relies on this without support.

    Normalization of Rac1 activity corrects ASD-like behaviors in a variety of animal models.

    “The fact that normalization of Rac1 activity corrects ASD-like behaviors in a variety of animal models suggests that targeting Rac1 may be a potential therapeutic approach for ASD.”
11 of 11

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Authors
Guo, D., Peng, Y., Wang, L., Sun, X., Wang, X., Liang, C., Yang, X., Li, S., Xu, J., Ye, W.-C., Jiang, B., & Shi, L.
Affiliations
  • Jinan University (CN)
  • Sun Yat-sen University (CN)
  • Zhejiang University (CN)
Corresponding author
  • Ye, Wen-Cai
  • Jiang, Bin
  • Shi, Lei
ORCID
Venue
Molecular Psychiatry
Publisher
Springer Science and Business Media LLC
Published
2019-08-06
Volume
26
Issue
5
Pages
1505-1519
ISSN
  • 1359-4184 (ISSN-L)
  • 1476-5578
DOI
10.1038/s41380-019-0472-7
PMC
PMC8159750
OpenAlex
W2965158022
Semantic Scholar
cedd6bfac86f82c00198fcb53150038fd72db66c
Cited by
102
References
66
Citations by year
2026: 6 · 2025: 16 · 2024: 25 · 2023: 13 · 2022: 20 · 2021: 15 · 2020: 7
FWCI
5.68
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Psychiatry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion, cc-by)
  • Rare & Special e-Zone (The Hong Kong University of Science and Technology) (submittedVersion)
Funders
  • Academy of Medical Sciences (AOMS-NAF0051003)
  • National Natural Science Foundation of China (81761130084, 81870869)
  • China Postdoctoral Science Foundation
  • Guangdong Science and Technology Department (2017BT01Y036)
  • The National Key R&D Program of China
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetics and Neurodevelopmental Disorders
  • Life Sciences › Neuroscience › Cognitive Neuroscience › Autism Spectrum Disorder Research
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
Keywords
autism spectrum disorder, DOCK4, Rac1, NMDA receptor, AMPA receptors, excitatory synaptic transmission, social behavior, social novelty preference, hippocampal CA1, synaptic dysfunction, protein synthesis, glutamate receptors, anxiety, spatial learning
MeSH terms
Autism Spectrum Disorder, Animals, Autistic Disorder, Hippocampus, Synaptic Transmission, Neuropeptides, Receptors, N-Methyl-D-Aspartate, Mice, Knockout, GTPase-Activating Proteins, rac1 GTP-Binding Protein, 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 2026-10-14.

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