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Autism gene Ube3a and seizures impair sociability by repressing VTA Cbln1

Nature volume 543, pages 507512 (23 March 2017) | Download Citation

Abstract

Maternally inherited 15q11-13 chromosomal triplications cause a frequent and highly penetrant type of autism linked to increased gene dosages of UBE3A, which encodes a ubiquitin ligase with transcriptional co-regulatory functions. Here, using in vivo mouse genetics, we show that increasing UBE3A in the nucleus downregulates the glutamatergic synapse organizer Cbln1, which is needed for sociability in mice. Epileptic seizures also repress Cbln1 and are found to expose sociability impairments in mice with asymptomatic increases in UBE3A. This Ube3a–seizure synergy maps to glutamate neurons of the midbrain ventral tegmental area (VTA), where Cbln1 deletions impair sociability and weaken glutamatergic transmission. We provide preclinical evidence that viral-vector-based chemogenetic activation of, or restoration of Cbln1 in, VTA glutamatergic neurons reverses the sociability deficits induced by Ube3a and/or seizures. Our results suggest that gene and seizure interactions in VTA glutamatergic neurons impair sociability by downregulating Cbln1, a key node in the expanding protein interaction network of autism genes.

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Acknowledgements

We thank O. DiStefano, G. Salimando and R. Broadhurst for colony work and the Harvard Medical School Neurobiology Imaging Facility (NINDS P30 Core Center Grant NS07203) and Boston Children’s Hospital IDDRC (1U54HD090255, P30HD18655). Supported by funding to V.K. (1R25NS070682 and an American Academy of Neurology Research Training Fellowship) and to M.P.A. from the NIH (1R01NS08916, 1R21MH100868, 1R21HD079249), Nancy Lurie Marks Family Foundation, Landreth Foundation, Simons Foundation, and Autism Speaks/National Alliance for Autism Research. R.A. was supported by Klarman Family Foundation.

Author information

Author notes

    • Vaishnav Krishnan
    • , David C. Stoppel
    •  & Yi Nong

    These authors contributed equally to this work.

Affiliations

  1. Department of Neurology, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, Massachusetts 02115, USA

    • Vaishnav Krishnan
    • , David C. Stoppel
    • , Yi Nong
    • , Mark A. Johnson
    • , Monica J. S. Nadler
    • , Ekim Ozkaynak
    • , Brian L. Teng
    • , Ikue Nagakura
    • , Michael A. Silva
    • , Sally Peterson
    • , Tristan J. Cruz
    •  & Matthew P. Anderson
  2. Department of Pathology, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, Massachusetts 02115, USA

    • David C. Stoppel
    • , Yi Nong
    • , Mark A. Johnson
    • , Monica J. S. Nadler
    • , Ekim Ozkaynak
    • , Brian L. Teng
    • , Ikue Nagakura
    • , Fahim Mohammad
    • , Michael A. Silva
    • , Sally Peterson
    • , Tristan J. Cruz
    • , Ramy Arnaout
    •  & Matthew P. Anderson
  3. Program in Neuroscience, Harvard Medical School, 300 Longwood Avenue, Boston, Massachusetts 02115, USA

    • David C. Stoppel
    •  & Matthew P. Anderson
  4. Department of Surgery, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, Massachusetts 02115, USA

    • Ekkehard M. Kasper
  5. Division of Clinical Informatics, Department of Internal Medicine, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, Massachusetts 02215, USA

    • Ramy Arnaout
  6. Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Ramy Arnaout
  7. Boston Children’s Hospital Intellectual and Developmental Disabilities Research Center, 300 Longwood Avenue, Boston, Massachusetts 02115, USA

    • Matthew P. Anderson

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Contributions

V.K., D.C.S., Y.N. and M.P.A. designed the study. V.K. and M.P.A wrote the manuscript. V.K., D.C.S. and Y.N. performed all experiments and analyses except for the following: E.O., M.J.S.N. and T.J.C validated microarray-identified gene regulations by qRT–PCR; F.M., R.A. and M.P.A. performed Gene Ontology and cluster visualization; I.N. performed nuclear/cytosolic fractionation studies; S.P. performed olfaction and open-field experiments; M.A.J., B.L.T., M.A.S., E.M.K. and M.J.S.N. developed molecular probes and constructs.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Matthew P. Anderson.

Reviewer Information Nature thanks E. Kim and the other anonymous reviewer(s) for their contribution to the peer review of this work.

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

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Figure 1

    This file shows the source gel for the representative bands provided in Extended Data Figure 2d.

Excel files

  1. 1.

    Supplementary Table 1

    This file lists genes identified as up or downregulated in cortical samples from Ube3a2x mice compared with WT littermates. A +/- 1.4 fold change cutoff was used. Accession numbers, fold change and p values are shown.

  2. 2.

    Supplementary Table 2

    This file lists microarray-identified genes by gene ontology classifications together with a calculated p value for enrichment within a specific biological process. Worksheet 1 within this table displays results of annotation through the Kyoto Encyclopedia of Genes and Genomes (KEGG). Worksheets 2-4 display the same results when gene lists were annotated by classifications as defined by the Gene Ontology Consortium (biological process, cellular component and molecular function).

  3. 3.

    Supplementary Table 3

    This file contains a list of selected 276 genes identified as strongly linked to autism from a larger list of 706 genes from the Simons Foundation Autism Research Initiative (SFARI, https://gene.sfari.org/autdb/HG_Home.do - see Basu, S. N., Kollu, R. & Banerjee-Basu, S. AutDB: a gene reference resource for autism research. Nucleic acids research 37, D832-836, doi:10.1093/nar/gkn835 (2009)).

  4. 4.

    Supplementary Table 4

    This file contains sequences for forward and reverse primers utilized in quantitative real-time PCR experiments.

  5. 5.

    Supplementary Table 5

    This file comprises: Worksheet 1 - A table of chamber times (in seconds) provided for each graph where "sniffing times" are depicted in the main and extended figures. This excludes Fig. 1d and Fig. 1h, where chamber times are depicted graphically in Extended Data Figs. 2e and 5b respectively; Worksheet 2 - Distances moved, chamber times and “sniffing times” (for "social" and "opposite") for a population of 101 wildtype seizure naïve FVB mice (males and females) with column statistics (mean and measures of variance) as well as results of Kolmogorov-Smirnov testing for the presence of a normal distribution; Worksheet 3 - Two-way repeated measures ANOVA analysis results for each graph depicting three chamber sociability data. The results of post-hoc testing are provided comparing "social" versus "opposite" sniffing times for each group separately.

  6. 6.

    Supplementary Table 6

    This file contains experimental data.

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DOI

https://doi.org/10.1038/nature21678

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