Deciphering cellular signals in adult mouse sinoatrial node cells
| Autor: | Lu Ren, Jessica L. Caldwell, G. R. Reddy, Phung N. Thai, Manuela Zaccolo, Julie Bossuyt, Crystal M. Ripplinger, Nipavan Chiamvimonvat, Yang Kevin Xiang, Madeline Nieves-Cintrón, Manuel F. Navedo |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
Cell signaling
Cell biology Science heart failure Bioengineering Biology Cardiovascular Article cAMP Ca2+/calmodulin-dependent protein kinase medicine 2.1 Biological and endogenous factors Aetiology Protein kinase A Cellular compartment Multidisciplinary Sinoatrial node cardiac cells biosensors Biology experimental methods medicine.anatomical_structure Förster resonance energy transfer Functional aspects of cell biology Second messenger system FRET Signal transduction signal transduction |
| Zdroj: | iScience, Vol 25, Iss 1, Pp 103693-(2022) iScience iScience, vol 25, iss 1 |
| ISSN: | 2589-0042 |
| Popis: | Summary Sinoatrial node (SAN) cells are the pacemakers of the heart. This study describes a method for culturing and infection of adult mouse SAN cells with FRET-based biosensors that can be exploited to examine signaling events. SAN cells cultured in media with blebbistatin or (S)-nitro-blebbistatin retain their morphology, protein distribution, action potential (AP) waveform, and cAMP dynamics for at least 40 h. SAN cells expressing targeted cAMP sensors show distinct β-adrenergic-mediated cAMP pools. Cyclic GMP, protein kinase A, Ca2+/CaM kinase II, and protein kinase D in SAN cells also show unique dynamics to different stimuli. Heart failure SAN cells show a decrease in cAMP and cGMP levels. In summary, a reliable method for maintaining adult mouse SAN cells in culture is presented, which facilitates studies of signaling networks and regulatory mechanisms during physiological and pathological conditions. Graphical abstract Highlights • Optimized culturing method of adult SAN cells from healthy and disease-state mice • Cultured adult SAN cells can be infected with diverse fluorescence biosensors • Examination of signaling networks, including discrete cAMP pools, in adult SAN cells • SAN cells cAMP and cGMP signals are impaired during heart failure Cell biology; Functional aspects of cell biology; Biology experimental methods |
| Databáze: | OpenAIRE |
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