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Got Milk? Identifying and Characterizing Lactation Defects in Genetically-Engineered Mouse Models

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Journal of Mammary Gland Biology and Neoplasia Aims and scope Submit manuscript

Abstract

The ability to produce and expel milk is important for the health and survival of all mammals. Nevertheless, our understanding of the molecular events underlying the execution of this process remains incomplete. Whilst impaired mammary gland development and lactational competence remains the subject of focused investigations, defects in these events may also be an unintended consequence of genetic manipulation in rodent models. In this technical report, we outline established and emerging methods to characterize lactation phenotypes in genetically-engineered mouse models. We discuss important considerations of common models, optimized conditions for mating and the importance of litter size and standardization. Methods for quantifying milk production and quality, as well as protocols for wholemount preparation, immunohistochemistry and the preparation of RNA and protein lysates are provided. This review is intended to help guide researchers new to the field of mammary gland biology in the systematic analysis of lactation defects and in the preparation of samples for more focused mechanistic investigations.

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Data Availability

Not applicable. All data generated or analysed during this study are included in this published article.

Abbreviations

CUBIC:

clear unobstructed brain imaging cocktails and computational analysis

d.p.c.:

days post coitum

E:

embryonic day

HIER:

heat-induced epitope retrieval

L:

lactation day

MG:

mammary gland

MMTV:

mouse mammary tumor virus

NBF:

neutral buffered formalin

PND:

post-natal day

TEB:

terminal end bud.

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Acknowledgements

We thank the UQ Biological Resource Facility staff for assistance with animal husbandry. We also thank animal facilities at the National Institutes of Environmental Health Sciences (NIEHS), University of Cambridge and University of Queensland for their advice and assistance in refining these protocols over the years, as well as Dr. Bethan Lloyd-Lewis (methyl green wholemount protocol) and Dr. Henrike Resemann (IHC protocol).

Funding

This work was supported by the National Health and Medical Research Council of Australia (1141008 and 1138214), the National Stem Cell Foundation of Australia and the Mater Foundation (Equity Trustees / AE Hingeley Trust).

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Authors and Affiliations

  1. Mater Research Institute-The University of Queensland, Faculty of Medicine, Woolloongabba, Queensland, 4102, Australia

    Teneale A. Stewart & Felicity M. Davis

  2. Translational Research Institute, Woolloongabba, Queensland, 4102, Australia

    Teneale A. Stewart & Felicity M. Davis

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  1. Teneale A. Stewart
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  2. Felicity M. Davis
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T.S. and F.D. prepared the manuscript.

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Correspondence to Felicity M. Davis.

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Animal Ethics

All animal work discussed and photographed in this manuscript was carried out in accordance with the Australian Code for the Care and Use of Animals for Scientific Purposes and the Queensland Animal care and Protection Act (2001), with local animal ethics approval. Mice were housed in individually-ventilated cages with food and water available ad libitum and a 12 h light cycle.

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Stewart, T.A., Davis, F.M. Got Milk? Identifying and Characterizing Lactation Defects in Genetically-Engineered Mouse Models. J Mammary Gland Biol Neoplasia 25, 255–272 (2020). https://doi.org/10.1007/s10911-020-09467-y

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