Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Behavioral, circuitry, and molecular aberrations by region-specific deficiency of the high-risk autism gene Cul3

Molecular Psychiatry volume 26, pages 1491–1504 (2021)Cite this article

Subjects

Abstract

Cullin 3 (Cul3) gene, which encodes a core component of the E3 ubiquitin ligase complex that mediates proteasomal degradation, has been identified as a true high-risk factor for autism. Here, by combining behavioral, electrophysiological, and proteomic approaches, we have examined how Cul3 deficiency contributes to the etiology of different aspects of autism. Heterozygous mice with forebrain Cul3 deletion displayed autism-like social interaction impairment and sensory-gating deficiency. Region-specific deletion of Cul3 leads to distinct phenotypes, with social deficits linked to the loss of Cul3 in prefrontal cortex (PFC), and stereotypic behaviors linked to the loss of Cul3 in striatum. Correlated with these behavioral alterations, Cul3 deficiency in forebrain or PFC induces NMDA receptor hypofunction, while Cul3 loss in striatum causes a cell type-specific alteration of neuronal excitability in striatal circuits. Large-scale profiling has identified sets of misregulated proteins resulting from Cul3 deficiency in different regions, including Smyd3, a histone methyltransferase involved in gene transcription. Inhibition or knockdown of Smyd3 in forebrain Cul3-deficient mice ameliorates social deficits and restores NMDAR function in PFC. These results have revealed for the first time a potential molecular mechanism underlying the manifestation of different autism-like behavioral deficits by Cul3 deletion in cortico-striatal circuits.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Data availability

Proteomic data will be deposited in a public repository. The access number and the dataset will be available for access.

References

  1. de la Torre-Ubieta L, Won H, Stein JL, Geschwind DH. Advancing the understanding of autism disease mechanisms through genetics. Nat Med. 2016;22:345–61.

    PubMed  PubMed Central  Google Scholar 

  2. De Rubeis S, He X, Goldberg AP, Poultney CS, Samocha K, Cicek AE, et al. Synaptic, transcriptional and chromatin genes disrupted in autism. Nature. 2014;515:209–15.

    PubMed  PubMed Central  Google Scholar 

  3. O’Roak BJ, Vives L, Girirajan S, Karakoc E, Krumm N, Coe BP, et al. Sporadic autism exomes reveal a highly interconnected protein network of de novo mutations. Nature. 2012;485:246–50.

    PubMed  PubMed Central  Google Scholar 

  4. Kong A, Frigge ML, Masson G, Besenbacher S, Sulem P, Magnusson G, et al. Rate of de novo mutations and the importance of father’s age to disease risk. Nature. 2012;488:471–5.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Stessman HA, Xiong B, Coe BP, Wang T, Hoekzema K, Fenckova M, et al. Targeted sequencing identifies 91 neurodevelopmental-disorder risk genes with autism and developmental-disability biases. Nat Genet. 2017;49:515–26.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Genschik P, Sumara I, Lechner E. The emerging family of CULLIN3-RING ubiquitin ligases (CRL3s): cellular functions and disease implications. EMBO J. 2013;32:2307–20.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Pinkas DM, Sanvitale CE, Bufton JC, Sorrell FJ, Solcan N, Chalk R, et al. Structural complexity in the KCTD family of Cullin3-dependent E3 ubiquitin ligases. Biochem J. 2017;474:3747–61.

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Dubiel W, Dubiel D, Wolf DA, Naumann M. Cullin 3-based ubiquitin ligases as master regulators of mammalian cell differentiation. Trends Biochem Sci. 2018;43:95–107.

    CAS  PubMed  Google Scholar 

  9. Chen HY, Liu CC, Chen RH. Cul3-KLHL20 ubiquitin ligase: physiological functions, stress responses, and disease implications. Cell Div. 2016;11:5.

    PubMed  PubMed Central  Google Scholar 

  10. Weiss LA, Shen Y, Korn JM, Arking DE, Miller DT, Fossdal R, et al. Association between microdeletion and microduplication at 16p11.2 and autism. N Engl J Med. 2008;358:667–75.

    CAS  PubMed  Google Scholar 

  11. Kumar RA, KaraMohamed S, Sudi J, Conrad DF, Brune C, Badner JA, et al. Recurrent 16p11.2 microdeletions in autism. Hum Mol Genet. 2008;17:628–38.

    CAS  PubMed  Google Scholar 

  12. McCarthy SE, Makarov V, Kirov G, Addington AM, McClellan J, Yoon S, et al. Microduplications of 16p11.2 are associated with schizophrenia. Nat Genet. 2009;41:1223–U1285.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Singer JD, Gurian-West M, Clurman B, Roberts JM. Cullin-3 targets cyclin E for ubiquitination and controls S phase in mammalian cells. Genes Dev. 1999;13:2375–87.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Ha S, Sohn IJ, Kim N, Sim HJ, Cheon KA. Characteristics of brains in autism spectrum disorder: structure, function and connectivity across the lifespan. Exp Neurobiol. 2015;24:273–84.

    PubMed  PubMed Central  Google Scholar 

  15. Hernandez LM, Rudie JD, Green SA, Bookheimer S, Dapretto M. Neural signatures of autism spectrum disorders: insights into brain network dynamics. Neuropsychopharmacology. 2015;40:171–89.

    PubMed  Google Scholar 

  16. Stoner R, Chow ML, Boyle MP, Sunkin SM, Mouton PR, Roy S, et al. Patches of disorganization in the neocortex of children with autism. N Engl J Med. 2014;370:1209–19.

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Schubert D, Martens GJ, Kolk SM. Molecular underpinnings of prefrontal cortex development in rodents provide insights into the etiology of neurodevelopmental disorders. Mol Psychiatry. 2015;20:795–809.

    CAS  PubMed  Google Scholar 

  18. Shepherd GM. Corticostriatal connectivity and its role in disease. Nat Rev Neurosci. 2013;14:278–91.

    CAS  PubMed  PubMed Central  Google Scholar 

  19. McEvoy JD, Kossatz U, Malek N, Singer JD. Constitutive turnover of cyclin E by Cul3 maintains quiescence. Mol Cell Biol. 2007;27:3651–66.

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Gulisano M, Broccoli V, Pardini C, Boncinelli E. Emx1 and Emx2 show different patterns of expression during proliferation and differentiation of the developing cerebral cortex in the mouse. Eur J Neurosci. 1996;8:1037–50.

    CAS  PubMed  Google Scholar 

  21. Gorski JA, Talley T, Qiu M, Puelles L, Rubenstein JL, Jones KR, et al. Cortical excitatory neurons and glia, but not GABAergic neurons, are produced in the Emx1-expressing lineage. J Neurosci. 2002;22:6309–14.

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Perry W, Minassian A, Lopez B, Maron L, Lincoln A. Sensorimotor gating deficits in adults with autism. Biol Psychiatry. 2007;61:482–6.

    PubMed  Google Scholar 

  23. Brumback AC, Ellwood IT, Kjaerby C, Iafrati J, Robinson S, Lee AT, et al. Identifying specific prefrontal neurons that contribute to autism-associated abnormalities in physiology and social behavior. Mol Psychiatry. 2017;23:2078–89.

    PubMed  PubMed Central  Google Scholar 

  24. 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.

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Tebartz van Elst L, Maier S, Fangmeier T, Endres D, Mueller GT, Nickel K, et al. Disturbed cingulate glutamate metabolism in adults with high-functioning autism spectrum disorder: evidence in support of the excitatory/inhibitory imbalance hypothesis. Mol Psychiatry. 2014;19:1314–25.

    CAS  PubMed  Google Scholar 

  26. Willsey AJ, Sanders SJ, Li M, Dong S, Tebbenkamp AT, Muhle RA, et al. Coexpression networks implicate human midfetal deep cortical projection neurons in the pathogenesis of autism. Cell. 2013;155:997–1007.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Ajram LA, Horder J, Mendez MA, Galanopoulos A, Brennan LP, Wichers RH, et al. Shifting brain inhibitory balance and connectivity of the prefrontal cortex of adults with autism spectrum disorder. Transl Psychiatry. 2017;7:e1137.

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Fuccillo MV. Striatal circuits as a common node for autism pathophysiology. Front Neurosci. 2016;10:27.

    PubMed  PubMed Central  Google Scholar 

  29. Burguiere E, Monteiro P, Mallet L, Feng G, Graybiel AM. Striatal circuits, habits, and implications for obsessive-compulsive disorder. Curr Opin Neurobiol. 2015;30:59–65.

    CAS  PubMed  Google Scholar 

  30. Langen M, Bos D, Noordermeer SD, Nederveen H, van Engeland H, Durston S. Changes in the development of striatum are involved in repetitive behavior in autism. Biol Psychiatry. 2014;76:405–11.

    PubMed  Google Scholar 

  31. Kreitzer AC, Malenka RC. Endocannabinoid-mediated rescue of striatal LTD and motor deficits in Parkinson’s disease models. Nature. 2007;445:643–7.

    CAS  PubMed  Google Scholar 

  32. Cepeda C, Andre VM, Yamazaki I, Wu N, Kleiman-Weiner M, Levine MS. Differential electrophysiological properties of dopamine D1 and D2 receptor-containing striatal medium-sized spiny neurons. Eur J Neurosci. 2008;27:671–82.

    PubMed  Google Scholar 

  33. Chen Y, Yang Z, Meng M, Zhao Y, Dong N, Yan H, et al. Cullin mediates degradation of RhoA through evolutionarily conserved BTB adaptors to control actin cytoskeleton structure and cell movement. Mol Cell. 2009;35:841–55.

    CAS  PubMed  Google Scholar 

  34. Zhang H, Macara IG. The PAR-6 polarity protein regulates dendritic spine morphogenesis through p190 RhoGAP and the Rho GTPase. Dev Cell. 2008;14:216–26.

    PubMed  PubMed Central  Google Scholar 

  35. Yan Z, Kim E, Datta D, Lewis DA, Soderling SH. Synaptic actin dysregulation, a convergent mechanism of mental disorders? J Neurosci. 2016;36:11411–7.

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Tashiro A, Minden A, Yuste R. Regulation of dendritic spine morphology by the rho family of small GTPases: antagonistic roles of Rac and Rho. Cereb Cortex. 2000;10:927–38.

    CAS  PubMed  Google Scholar 

  37. Quassollo G, Wojnacki J, Salas DA, Gastaldi L, Marzolo MP, Conde C, et al. A RhoA signaling pathway regulates dendritic golgi outpost formation. Curr Biol. 2015;25:971–82.

    CAS  PubMed  Google Scholar 

  38. Lin GN, Corominas R, Lemmens I, Yang X, Tavernier J, Hill DE, et al. Spatiotemporal 16p11.2 protein network implicates cortical late mid-fetal brain development and KCTD13-Cul3-RhoA pathway in psychiatric diseases. Neuron. 2015;85:742–54.

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Shulha HP, Cheung I, Whittle C, Wang J, Virgil D, Lin CL, et al. Epigenetic signatures of autism: trimethylated H3K4 landscapes in prefrontal neurons. Arch Gen Psychiatry. 2012;69:314–24.

    CAS  PubMed  Google Scholar 

  40. Peserico A, Germani A, Sanese P, Barbosa AJ, Di Virgilio V, Fittipaldi R, et al. A SMYD3 small-molecule inhibitor impairing cancer cell growth. J Cell Physiol. 2015;230:2447–60.

    CAS  PubMed  PubMed Central  Google Scholar 

  41. Amodio DM, Frith CD. Meeting of minds: the medial frontal cortex and social cognition. Nat Rev Neurosci. 2006;7:268–77.

    CAS  PubMed  Google Scholar 

  42. Qin L, Ma K, Wang ZJ, Hu Z, Matas E, Wei J, et al. Social deficits in Shank3-deficient mouse models of autism are rescued by histone deacetylase (HDAC) inhibition. Nat Neurosci. 2018;21:564–75.

    CAS  PubMed  PubMed Central  Google Scholar 

  43. Peca J, Feliciano C, Ting JT, Wang W, Wells MF, Venkatraman TN, et al. Shank3 mutant mice display autistic-like behaviours and striatal dysfunction. Nature. 2011;472:437–42.

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Won H, Lee HR, Gee HY, Mah W, Kim JI, Lee J, et al. Autistic-like social behaviour in Shank2-mutant mice improved by restoring NMDA receptor function. Nature. 2012;486:261–5.

    CAS  PubMed  Google Scholar 

  45. Wang W, Rein B, Zhang F, Tan T, Zhong P, Qin LY, et al. Chemogenetic activation of prefrontal cortex rescues synaptic and behavioral deficits in a mouse model of 16p11.2 deletion syndrome. J Neurosci. 2018;38:5939–48.

    CAS  PubMed  PubMed Central  Google Scholar 

  46. Escamilla CO, Filonova I, Walker AK, Xuan ZX, Holehonnur R, Espinosa F, et al. Kctd13 deletion reduces synaptic transmission via increased RhoA. Nature. 2017;551:227–31.

    CAS  PubMed  PubMed Central  Google Scholar 

  47. McGourty CA, Akopian D, Walsh C, Gorur A, Werner A, Schekman R, et al. Regulation of the CUL3 ubiquitin ligase by a calcium-dependent co-adaptor. Cell. 2016;167:525–38 e514.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Xiaoqing Chen for excellent technical support. We acknowledge the support of New York State Center of Excellence in Bioinformatics and Life Sciences and University at Buffalo. This work was supported by: Nancy Lurie Marks Family Foundation (Z.Y.), National Institutes of Health (MH112237 and MH108842) (Z.Y.), Brain and Behavior Foundation (24910) (M.R.) and National Center for Advancing Translational Sciences of the National Institutes of Health (KL2TR001413) (M.R.).

Author information

Author notes

  1. These authors contributed equally: Maximiliano Rapanelli, Tao Tan

Authors and Affiliations

  1. Department of Physiology and Biophysics, Jacobs School of Medicine and Biomedical Sciences, State University of New York at Buffalo, Buffalo, NY, USA

    Maximiliano Rapanelli, Tao Tan, Wei Wang, Zi-Jun Wang, Ping Zhong, Luye Qin, Kaijie Ma & Zhen Yan

  2. New York State Center of Excellence in Bioinformatics and Life Sciences, Buffalo, NY, USA

    Xue Wang & Jun Qu

  3. Hunter James Kelly Research Institute, Department of Neurology, State University of New York at Buffalo, Buffalo, NY, USA

    Luciana Frick

Authors
  1. Maximiliano Rapanelli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tao Tan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xue Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zi-Jun Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ping Zhong
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Luciana Frick
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Luye Qin
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Kaijie Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Jun Qu
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Zhen Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

M.R. performed the behavioral, immunocytochemical, anatomical, and some biochemical experiments, carried out bioinformatic analyses, and wrote the draft. T.T., W.W. and P.Z. performed electrophysiological experiments and analyzed data. X.W. and J.Q. carried out proteomic experiments and analyzed the data. Z.W., L.F., L.Q. and K.M. participated in the biochemical, immunocytochemical, anatomical, and behavioral experiments. Z.Y. designed the experiments, supervised the project, and wrote the paper.

Corresponding author

Correspondence to Zhen Yan.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Rapanelli, M., Tan, T., Wang, W. et al. Behavioral, circuitry, and molecular aberrations by region-specific deficiency of the high-risk autism gene Cul3. Mol Psychiatry 26, 1491–1504 (2021). https://doi.org/10.1038/s41380-019-0498-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41380-019-0498-x

This article is cited by

Search

Advanced search

Quick links