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Methods for imaging intracellular calcium signals in the mouse mammary epithelium in 2- and 3-dimensions

Folacci, M.; Chalmers, S. B.; Davis, F. M.

2024-03-01 physiology
10.1101/2024.02.27.582265 bioRxiv
Show 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 visualise 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.

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