Methods for Imaging Intracellular Calcium Signals in the Mouse Mammary Epithelium in Two and Three Dimensions

Mathilde Folacci; Silke B Chalmers; Felicity M Davis

doi:10.1007/978-1-0716-4164-4_15

Methods for Imaging Intracellular Calcium Signals in the Mouse Mammary Epithelium in Two and Three Dimensions

Methods Mol Biol. 2025:2861:195-212. doi: 10.1007/978-1-0716-4164-4_15.

Authors

Mathilde Folacci¹, Silke B Chalmers¹, Felicity M Davis^{2

3

4

5

6}

Affiliations

¹ Department of Biomedicine, Aarhus University, Aarhus, Denmark.
² Department of Biomedicine, Aarhus University, Aarhus, Denmark. [email protected].
³ School of Biomedical Sciences, University of New South Wales, Sydney, NSW, Australia. [email protected].
⁴ Aarhus Institute of Advanced Studies, Aarhus University, Aarhus, Denmark. [email protected].
⁵ Danish Research Institute of Translational Neuroscience, Aarhus University, Aarhus, Denmark. [email protected].
⁶ School of Pharmacy, University of Queensland, Brisbane, QLD, Australia. [email protected].

PMID: 39395107
DOI: 10.1007/978-1-0716-4164-4_15

Abstract

The mammary gland has a central role in optimal mammalian development and survival. Contractions of smooth muscle-like basal (or myoepithelial) cells in the functionally mature mammary gland in response to oxytocin are essential for milk ejection and are tightly regulated by intracellular calcium (Ca²⁺). Using mice expressing a genetically encoded Ca²⁺ indicator (GCaMP6f), we present in this chapter a method to visualize at high spatiotemporal resolution changes in intracellular Ca²⁺ in mammary epithelial cells, both in vitro (2D) and ex vivo (3D). The procedure to optimally prepare mammary tissue and primary cells is presented in detail.

Keywords: Calcium imaging; Calcium signaling; Gcamp6f; Genetically encoded ca2+ indicator; Lactation; Mammary epithelial cells; Mammary gland; Oxytocin.

MeSH terms

Animals
Calcium Signaling*
Calcium* / metabolism
Epithelial Cells / metabolism
Epithelium / metabolism
Female
Imaging, Three-Dimensional / methods
Mammary Glands, Animal* / cytology
Mammary Glands, Animal* / metabolism
Mice

Substances

Calcium