Got Milk? Identifying and Characterizing Lactation Defects in Genetically-Engineered Mouse Models
- PDF / 5,857,172 Bytes
- 18 Pages / 595.276 x 790.866 pts Page_size
- 102 Downloads / 160 Views
Got Milk? Identifying and Characterizing Lactation Defects in Genetically-Engineered Mouse Models Teneale A. Stewart 1,2 & Felicity M. Davis 1,2 Received: 21 July 2020 / Accepted: 27 October 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
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. Keywords Mammary gland . Lactation . Secretion . Development . Genetically-engineered mouse model . Mammalian reproduction
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.
* Felicity M. Davis [email protected] 1
Mater Research Institute-The University of Queensland, Faculty of Medicine, Woolloongabba, Queensland 4102, Australia
2
Translational Research Institute, Woolloongabba, Queensland 4102, Australia
Introduction The development of novel genetically-engineered mouse models—fueled by advancements in methods for rapid and efficient genome editing [1]—can yield unexpected but fascinating phenotypes [2]. This may occur, for example, in an unanticipated organ using a global gene deletion strategy. It may also occur in a “tissue-specific” knockout model, e.g., a Cre/lox system [3], where Cre-recombinase expression is under the control of a more widely-expressed gene promoter (e.g., Fig. 1). If genetic deletion occurs in the mammary gland, it may result in an inability of the female parent to adequately feed her offspring. This may be identified by early cannibalization, stunted postnatal growth and/or the eventual loss of the entire litter. Although this phenotype may, at first, be a frustrating and progress-limiting delay to the original research question, it also represents a wonderful opportunity to characterize a fundamental aspect of mammalian biology. It is with this in mind that the cur
Data Loading...
