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Illuminating cell signaling with genetically encoded FRET biosensors in adult mouse cardiomyocytes.
- Source :
-
Journal of General Physiology . Nov2018, Vol. 150 Issue 11, p1567-1582. 16p. - Publication Year :
- 2018
-
Abstract
- FRET-based biosensor experiments in adult cardiomyocytes are a powerful way of dissecting the spatiotemporal dynamics of the complicated signaling networks that regulate cardiac health and disease. However, although much information has been gleaned from FRET studies on cardiomyocytes from larger species, experiments on adult cardiomyocytes from mice have been difficult at best. Thus the large variety of genetic mouse models cannot be easily used for this type of study. Here we develop cell culture conditions for adult mouse cardiomyocytes that permit robust expression of adenoviral FRET biosensors and reproducible FRET experimentation. We find that addition of 6.25 μM blebbistatin or 20 μM (S)-nitroblebbistatin to a minimal essential medium containing 10 mM HEP ES and 0.2% BSA maintains morphology of cardiomyocytes from physiological, pathological, and transgenic mouse models for up to 50 h after adenoviral infection. This provides a 10-15-h time window to perform reproducible FRET readings using a variety of CFP/YFP sensors between 30 and 50 h postinfection. The culture is applicable to cardiomyocytes isolated from transgenic mouse models as well as models with cardiac diseases. Therefore, this study helps scientists to disentangle complicated signaling networks important in health and disease of cardiomyocytes. [ABSTRACT FROM AUTHOR]
- Subjects :
- *CELL communication
*BIOSENSORS
*HEART cells
*HEART cell culture
*BLEBBISTATIN
Subjects
Details
- Language :
- English
- ISSN :
- 00221295
- Volume :
- 150
- Issue :
- 11
- Database :
- Academic Search Index
- Journal :
- Journal of General Physiology
- Publication Type :
- Academic Journal
- Accession number :
- 132942034
- Full Text :
- https://doi.org/10.1085/jgp.201812119