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A guide to designing germline-dependent epigenetic inheritance experiments in mammals

Abstract

Recent work has demonstrated that environmental factors experienced by parents can affect their offspring across multiple generations, and that such transgenerational transmission can depend on the germline. Causal evidence for the involvement of germ cells is rare, however, and the underlying molecular mechanisms remain poorly understood. Further, studies often employ varying methods in experimental design and data interpretation. We provide a critical analysis of these issues and suggest possible solutions and guidelines for improving study design and generating reproducible and high-quality data.

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Figure 1: Experimental strategies to causally test germline dependence of inherited phenotypes.
Figure 2: Weaning strategies for rodent studies of germline epigenetic inheritance.

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Acknowledgements

The lab of IMM is funded by the University of Zurich, the ETH Zurich, the Swiss National Science Foundation, the ETHZ Foundation, Roche and private sponsors. J.B. received funding from the Forschungskredit of the University of Zurich (grant no. FK-15-035), the Vontobel Foundation, the Betty and David Koetser Foundation for Brain Research, and the EMDO Foundation. We thank G. van Steenwyk for critical reading of the manuscript and S. Steinbacher for illustrations.

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Integrated supplementary information

Supplementary Figure 1 Male germ cell development and spermatogenesis in mice.

Male germ cell development begins prenatally and continues throughout life in the testes. Epigenetic modifications involved in germline epigenetic inheritance can be studied in mature sperm cells that are stored for release in the cauda epididymis (A). Epigenetic modifications can already be induced and detected during early developmental stages, affecting primordial germ cells (PGCs, B) and/or spermatogonial stem cells (SSCs, C). Environmental factors can also impact Sertoli cells (D) or the epididymal duct (E), thus potentially affecting developing sperm cells upon transit through these structures. To unveil the mechanisms of germline epigenetic inheritance, future studies should aim to identify epigenetic modifications in some of these structures. SPCs = spermatocytes; PL=Pre-leptotene; P=Pachytene

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Bohacek, J., Mansuy, I. A guide to designing germline-dependent epigenetic inheritance experiments in mammals. Nat Methods 14, 243–249 (2017). https://doi.org/10.1038/nmeth.4181

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