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Methods for Imaging Intracellular Calcium Signals in the Mouse Mammary Epithelium in Two and Three Dimensions
Mathilde Folacci1, Silke B Chalmers1, Felicity M Davis2,3,4,5,6
1Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Methods in Molecular Biology (Clifton, N.J.)
|October 12, 2024
Summary
Researchers developed a new method to visualize calcium signaling in mammary gland cells. This technique uses a genetically encoded calcium indicator in mice to observe intracellular calcium changes crucial for milk ejection.
Area of Science:
- Mammalian developmental biology
- Cellular physiology
- Biophysics
Background:
- The mammary gland is vital for mammalian development and survival.
- Milk ejection relies on oxytocin-induced contractions of myoepithelial cells.
- Intracellular calcium (Ca2+) tightly regulates these contractions.
Purpose of the Study:
- To present a method for visualizing intracellular Ca2+ dynamics in mammary epithelial cells.
- To enable high spatiotemporal resolution imaging of Ca2+ changes.
- To detail the preparation of mammary tissue and primary cells for imaging.
Main Methods:
- Utilized genetically encoded Ca2+ indicator (GCaMP6f) in mice.
- Developed protocols for both in vitro (2D) and ex vivo (3D) imaging.
- Optimized mammary tissue and primary cell preparation techniques.
Main Results:
- Successfully visualized intracellular Ca2+ changes in mammary epithelial cells.
- Achieved high spatiotemporal resolution in imaging Ca2+ dynamics.
- Provided detailed procedures for tissue and cell preparation.
Conclusions:
- The presented method allows detailed study of Ca2+ signaling in mammary gland function.
- This technique is valuable for understanding the regulation of milk ejection.
- The detailed protocols facilitate reproducibility and further research in mammary gland physiology.
