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Tamoxifen induction of Cre recombinase does not cause long-lasting or sexually divergent responses in the CNS epigenome or transcriptome: implications for the design of aging studies

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Abstract

The systemic delivery of tamoxifen (Tam) to activate inducible CreERT2-loxP transgenic mouse systems is now widely used in neuroscience studies. This critical technological advancement allows temporal control of DNA-cre recombination, avoidance of embryonically lethal phenotypes, and minimization of residual cell labeling encountered in constitutively active drivers. Despite its advantages, the use of Tam has the potential to cause long-lasting, uncharacterized side effects on the transcriptome and epigenome in the CNS, given its mixed estrogen receptor (ER) agonist/antagonist actions. With the welcome focus on including both sexes in biomedical studies and efforts to understand sex differences, Tam administration could also cause sexually divergent responses that would confound studies. To examine these issues, epigenetic and transcriptomic profiles were compared in C57BL/6 J female and male hippocampus, cortex, and retina 1 month after a 5-day Tam treatment typical for cre induction, or vehicle control (sunflower seed oil). Cytosine methylation and hydroxymethylation levels, in both CG and non-CG contexts, were unchanged as determined by oxidative bisulfite sequencing. Long-lasting Tam transcriptomic effects were also not evident/minimal. Furthermore, there is no evidence of sexually divergent responses with Tam administration and Tam did not alter sex differences evident in controls. Combined with recently reported data that Tam alone does not cause long-lasting changes in behavior and neurogenesis, our findings provide confidence that Tam can be used as a cre-recombinase inducer without introducing significant confounds in transcriptomic and epigenomic neuroscience studies, particularly those focused on genomic and transcriptomic aspects of the aging brain.

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Acknowledgements

The authors acknowledge the Laboratory for Molecular Biology and Cytometry Research at OUHSC for the use of the Core Facility which provided Illumina MiSeq Next Generation Sequencing services and the Oklahoma Medical Research Foundation Clinical Genomics Center which provided NextSeq services Computing for this project was performed at the OU Supercomputing Center for Education and Research (OSCER) at the University of Oklahoma (OU).

Support

This work was supported by grants from the National Institutes of Health (NIH) P30AG050911, R56AG059430, R01AG58430, P20GM125528, R01AG0256161, Veterans Affairs I01BX003906, Oklahoma Center for Adult Stem Cell Research (OCASCR) grant through the Oklahoma Tobacco Settlement Endowment Trust, and Presbyterian Health Foundation.

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  1. Ana J. Chucair-Elliott and Sarah R. Ocanas contributed equally to this work.

Authors and Affiliations

  1. Reynolds Oklahoma Center on Aging, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Ana J. Chucair-Elliott, Sarah R. Ocanas, David R. Stanford, Niran Hadad, Benjamin Wronowski, Laura Otalora, Michael B. Stout & Willard M. Freeman

  2. Department of Physiology, University of Oklahoma Health Sciences Center, 975 NE 10th Street, Oklahoma City, OK, 73104, USA

    Ana J. Chucair-Elliott, Sarah R. Ocanas, David R. Stanford, Laura Otalora & Willard M. Freeman

  3. Oklahoma Nathan Shock Center for Aging, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Niran Hadad & Willard M. Freeman

  4. Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Niran Hadad & Willard M. Freeman

  5. Department of Nutritional Sciences, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Michael B. Stout

  6. Oklahoma City Veterans Affairs Medical Center, Oklahoma City, OK, USA

    Willard M. Freeman

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Chucair-Elliott, A.J., Ocanas, S.R., Stanford, D.R. et al. Tamoxifen induction of Cre recombinase does not cause long-lasting or sexually divergent responses in the CNS epigenome or transcriptome: implications for the design of aging studies. GeroScience 41, 691–708 (2019). https://doi.org/10.1007/s11357-019-00090-2

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