Abstract

Ten-eleven translocation (TET) enzymes mediate the conversion of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), which is enriched in brain, and its ultimate DNA demethylation. However, the influence of TET and 5hmC on gene transcription in brain remains elusive. We found that ten-eleven translocation protein 1 (TET1) was downregulated in mouse nucleus accumbens (NAc), a key brain reward structure, by repeated cocaine administration, which enhanced behavioral responses to cocaine. We then identified 5hmC induction in putative enhancers and coding regions of genes that have pivotal roles in drug addiction. Such induction of 5hmC, which occurred similarly following TET1 knockdown alone, correlated with increased expression of these genes as well as with their alternative splicing in response to cocaine administration. In addition, 5hmC alterations at certain loci persisted for at least 1 month after cocaine exposure. Together, these reveal a previously unknown epigenetic mechanism of cocaine action and provide new insight into how 5hmC regulates transcription in brain in vivo.

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Acknowledgements

We thank A. Chess for critical comments and O. Jabado and M. Mahajan from the Mount Sinai Genomics Core for technical support. This work was supported by grants from the National Institute on Drug Abuse (E.J.N.), the National Institutes of Health (P.J., P.C., G.F., K.F.F. and H.S.) and the Simons Foundation (H.S.).

Author information

Author notes

    • Ningyi Shao
    • , Keith E Szulwach
    •  & Vincent Vialou

    These authors contributed equally to this work.

Affiliations

  1. Fishberg Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA.

    • Jian Feng
    • , Ningyi Shao
    • , Vincent Vialou
    • , Jimmy Huynh
    • , Deveroux Ferguson
    • , Michael E Cahill
    • , Ja Wook Koo
    • , Efrain Ribeiro
    • , Benoit Labonte
    • , Benjamin M Laitman
    • , David Estey
    • , Victoria Stockman
    • , Pamela Kennedy
    • , Isaac Mensah
    • , Patrizia Casaccia
    • , Li Shen
    •  & Eric J Nestler
  2. Department of Human Genetics, Emory University School of Medicine, Atlanta, Georgia, USA.

    • Keith E Szulwach
    • , Yujing Li
    •  & Peng Jin
  3. Institut National de la Santé et de la Recherhe Médicale (INSERM) U1130, CNRS UMR8246, UPMC UM18, Neuroscience Paris Seine, Paris, France.

    • Vincent Vialou
    •  & Thomas Couroussé
  4. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.

    • Chun Zhong
    • , Guo-li Ming
    •  & Hongjun Song
  5. Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, California, USA.

    • Thuc Le
    •  & Guoping Fan
  6. The McGill Group for Suicide Studies, Douglas Hospital Research Centre, McGill University, Montreal, Canada.

    • Gustavo Turecki
  7. Pasarow Mass Spectrometry Laboratory, Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, California, USA.

    • Kym F Faull

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Contributions

The studies were conceived and designed by E.J.N., P.J. and J.F. J.F. performed RNA-seq and ChIP-seq. K.E.S., Y.L. and J.F. performed 5hmC capture and sequencing. L.S., N.S. and J.F. performed bioinformatic analyses. J.F. and J.H. performed oxBS-seq. JF., V.V., B.M.L., V.S. and I.M. performed qPCR analyses. V.V., D.E., T.C. and J.F. performed immunohistochemistry. J.F., V.V., D.F., J.K. and E.R. performed stereotaxic surgeries and behavioral assays. T.L., K.F.F. and G.F. contributed LC-ESI-MS/MS data. C.Z., G.M. and H.S. provided AAV-Tet1 shRNA and AAV-TET1 viruses. M.E.C. performed western blotting. P.C. contributed reagents. G.T. contributed human samples. D.F., B.L., B.M.L., V.S. and P.K. helped to prepare the samples and collect the data. The paper was written by J.F. and E.J.N. and was edited by the other authors.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Eric J Nestler.

Integrated supplementary information

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    Supplementary Text and Figures

    Supplementary Figures 1–8 and Supplementary Tables 1, 5 and 7

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    Supplementary Methods Checklist

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  1. 1.

    Supplementary Table 2

    Gene regions that display cocaine-induced changes in 5hmC

  2. 2.

    Supplementary Table 3

    Genes whose putative nearby enhancers show cocaine-induced switches to chromatin state 4 or 5

  3. 3.

    Supplementary Table 4

    Gene ontology of genes whose putative nearby enhancers show cocaine-induced switches to chromatin states 4 or 5

  4. 4.

    Supplementary Table 6

    Genes that show overlap in cocaine regulation of 5hmC and alternative splicing

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DOI

https://doi.org/10.1038/nn.3976

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