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Chronically high stress hormone levels dysregulate sperm long noncoding RNAs and their embryonic microinjection alters development and affective behaviours

Molecular Psychiatry (2023)Cite this article

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Abstract

Previous studies on paternal epigenetic inheritance have shown that sperm RNAs play a role in this type of inheritance. The microinjection of sperm small noncoding RNAs into fertilised mouse oocytes induces reprogramming of the early embryo, which is thought to be responsible for the differences observed in adult phenotype. While sperm long noncoding RNAs (lncRNAs) have also been investigated in a previous study, their microinjection into fertilised oocytes did not yield conclusive results regarding their role in modulating brain development and adult behavioural phenotypes. Therefore, in the current study we sought to investigate this further. We used our previously established paternal corticosterone (stress hormone) model to assess sperm lncRNA expression using CaptureSeq, a sequencing technique that is more sensitive than the ones used in other studies in the field. Paternal corticosterone exposure led to dysregulation of sperm long noncoding RNA expression, which encompassed lncRNAs, circular RNAs and transposable element transcripts. Although they have limited functional annotation, bioinformatic approaches indicated the potential of these lncRNAs in regulating brain development and function. We then separated and isolated the sperm lncRNAs and performed microinjections into fertilised oocytes, to generate embryos with modulated lncRNA populations. We observed that the resulting adult offspring had lower body weight and altered anxiety and affective behavioural responses, demonstrating roles for lncRNAs in modulating development and brain function. This study provides novel insights into the roles of lncRNAs in epigenetic inheritance, including impacts on brain development and behaviours of relevance to affective disorders.

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Fig. 1: Long noncoding RNA capture sequencing data analysis.
Fig. 2: Investigation of other long RNA types, and cis-target prediction.
Fig. 3: Analysis of transposable elements in the promoter regions of up and down regulated lncRNAs.
Fig. 4: Experimental design and offspring body weight.
Fig. 5: Assessment of anxiety- and depressive-like behaviours.
Fig. 6: Assessment of social behaviours.

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Data availability

The sequencing datasets generated and analysed during the current study have been deposited in the European Nucleotide Archive (ENA) repository at EMBL‐EBI under accession number PRJEB66376 (https://www.ebi.ac.uk/ena/browser/view/PRJEB66376). All other relevant data not included in the published article will be made available upon reasonable request.

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Acknowledgements

We thank Brett Purcell and Travis Featherby for their assistance with behavioural data collection, Craig Thomson and Maria Bastias for their support with animal housing, Fiona Waters for performing the microinjections, the staff at the WEHI animal facility, and the Spartan High Performance Computing resource offered by the University of Melbourne. LBH is supported by the Melbourne Research Scholarship. AJH, TWB and TYP were supported by NHMRC Project Grant funding. AJH also received support from a NHMRC Principal Research Fellowship and a DHB Foundation (Equity Trustees) Grant. The Florey Institute of Neuroscience and Mental Health acknowledges the support from the Victorian Government’s Operational Infrastructure Support Grant.

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Author notes

  1. These authors contributed equally: T. Y. Pang, A. J. Hannan.

Authors and Affiliations

  1. Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, VIC, Australia

    L. B. Hoffmann, T. Y. Pang & A. J. Hannan

  2. Florey Department of Neuroscience and Mental Health, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, Australia

    L. B. Hoffmann, B. Li, T. Y. Pang & A. J. Hannan

  3. Cognitive Neuroepigenetics Laboratory, Queensland Brain Institute, The University of Queensland, Brisbane, QLD, Australia

    Q. Zhao, W. Wei, L. J. Leighton & T. W. Bredy

  4. Department of Anatomy and Physiology, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, Australia

    A. J. Hannan

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Contributions

LBH contributed to planning experiments, conducted all experiments and analyses, and wrote the manuscript. BL assisted in conducting animal behavioural studies. QZ and WW performed the CaptureSeq gene expression analysis. LJL conducted the capture sequencing. TWB supervised the CaptureSeq and gave critical feedback to its analysis. TYP envisioned the study, supervised study design, data collection and statistical analyses, and reviewed the manuscript. AJH envisioned and funded the study, contributed to planning experiments, provided critical feedback throughout the experiments, and reviewed and edited the manuscript.

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Correspondence to A. J. Hannan.

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Hoffmann, L.B., Li, B., Zhao, Q. et al. Chronically high stress hormone levels dysregulate sperm long noncoding RNAs and their embryonic microinjection alters development and affective behaviours. Mol Psychiatry (2023). https://doi.org/10.1038/s41380-023-02350-2

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