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Tet3 ablation in adult brain neurons increases anxiety-like behavior and regulates cognitive function in mice

Molecular Psychiatry volume 26, pages 1445–1457 (2021)Cite this article

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Abstract

TET3 is a member of the ten-eleven translocation (TET) family of enzymes which oxidize 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC). Tet3 is highly expressed in the brain, where 5hmC levels are most abundant. In adult mice, we observed that TET3 is present in mature neurons and oligodendrocytes but is absent in astrocytes. To investigate the function of TET3 in adult postmitotic neurons, we crossed Tet3 floxed mice with a neuronal Cre-expressing mouse line, Camk2a-CreERT2, obtaining a Tet3 conditional KO (cKO) mouse line. Ablation of Tet3 in adult mature neurons resulted in increased anxiety-like behavior with concomitant hypercorticalism, and impaired hippocampal-dependent spatial orientation. Transcriptome and gene-specific expression analysis of the hippocampus showed dysregulation of genes involved in glucocorticoid signaling pathway (HPA axis) in the ventral hippocampus, whereas upregulation of immediate early genes was observed in both dorsal and ventral hippocampal areas. In addition, Tet3 cKO mice exhibit increased dendritic spine maturation in the ventral CA1 hippocampal subregion. Based on these observations, we suggest that TET3 is involved in molecular alterations that govern hippocampal-dependent functions. These results reveal a critical role for epigenetic modifications in modulating brain functions, opening new insights into the molecular basis of neurological disorders.

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Fig. 1: TET3 is present in mature neurons and diminished in the brain of Tet3 cKO mice, with no alteration in 5hmC levels.
Fig. 2: Tet3 cKO mice showed increased anxiety-like behavior and corticosterone levels.
Fig. 3: Tet3 cKO mice showed spatial orientation impairment, but normal recognition memory.
Fig. 4: Transcriptome analysis showed a predominant alteration in transcript levels in the ventral hippocampus.
Fig. 5: Three-dimensional morphometric analysis of Golgi-impregnated neurons of the CA1 hippocampus reveals an increase in dendritic spine maturation in Tet3 cKO mice.

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Acknowledgements

We thank Julian Peat and Christel Krueger (Babraham Institute, UK) for help with the Tet3 floxed mice, João Sobral (Genomics Unit, IGC, Portugal) for RNA-Seq analysis, and Daniel Neves and Daniel Sobral (Bioinformatics Unit, IGC, Portugal) for Bioinformtaic analysis. This work was supported by National Funds through Foundation for Science and Technology (FCT) fellowships (PD/BD/106049/2015 to CA, PD/BD/128074/2016 to JDS, IF/01079/2014 to LP, SFRH/BD/101298/2014 to SGG, SFRH/BD/131278/2017 to ELC, and IF/00047/2012 and CEECIND/00371/2017 to CJM); FCT project grant (PTDC/BIA-BCM/121276/2010) to CJM; EpiGeneSys Small Collaborative project to LP and MRB; BIAL Foundation Grant 427/14 to LP; Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER; NORTE-01-0145-FEDER-000013); FEDER funds, through the Competitiveness Factors Operational Programme (COMPETE), and National Funds, through the FCT (POCI-01-0145-FEDER-007038).

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  1. These authors contributed equally: Luísa Pinto, C. Joana Marques

Authors and Affiliations

  1. Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057, Braga, Portugal

    Cláudia Antunes, Jorge D. Da Silva, Sónia Guerra-Gomes, Nuno D. Alves, Fábio Ferreira, Eduardo Loureiro-Campos, Nuno Sousa & Luísa Pinto

  2. ICVS/3B’s—PT Government Associate Laboratory, 4710-057, Braga/Guimarães, Portugal

    Cláudia Antunes, Jorge D. Da Silva, Sónia Guerra-Gomes, Nuno D. Alves, Fábio Ferreira, Eduardo Loureiro-Campos, Nuno Sousa & Luísa Pinto

  3. Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, E1 2AT, UK

    Miguel R. Branco

  4. Epigenetics Programme, The Babraham Institute, Cambridge, CB22 3AT, UK

    Wolf Reik

  5. The Wellcome Trust Sanger Institute, Cambridge, CB10 1SA, UK

    Wolf Reik

  6. Department of Genetics, Faculty of Medicine, University of Porto (FMUP), 4200-319, Porto, Portugal

    C. Joana Marques

  7. i3S—Instituto de Investigação e Inovação em Saúde, Universidade do Porto, 4200-135, Porto, Portugal

    C. Joana Marques

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Contributions

CA designed the study, performed the experiments, analyzed the data, and wrote the manuscript; JDS performed gene ontology analysis of QuantSeq results, statistical analysis, and wrote the manuscript. SGG and NDA helped with the behavioral tests and respective analysis. ELC helped with the behavioral tests. FF helped with neuronal morphology analysis. MRB helped with RNA-Seq data. NS organized and wrote the manuscript; WR contributed with the Tet3 conditional mouse strain; LP and CJM designed the study, organized, and wrote the manuscript. All authors revised and approved the final manuscript.

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Correspondence to Luísa Pinto or C. Joana Marques.

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Antunes, C., Da Silva, J.D., Guerra-Gomes, S. et al. Tet3 ablation in adult brain neurons increases anxiety-like behavior and regulates cognitive function in mice. Mol Psychiatry 26, 1445–1457 (2021). https://doi.org/10.1038/s41380-020-0695-7

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