Year: 2021
Type: article
Abstract: 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, Ca 2+ /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.
Source: iScience
Language: English
Subfield: Cardiology and Cardiovascular Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $3,000
Funders: U.S. Department of Veterans Affairs, American Heart Association, University of California Institute for Mexico and the United States, British Heart Foundation, Consejo Nacional de Ciencia y Tecnología, National Institutes of Health, University of California, Davis, National Heart, Lung, and Blood Institute, NHLBI Division of Intramural Research
Awards: G1066778374, Coupling of vascular CaV1.2 channels in health and disease, G1508866, G1780951393, G1959900319, G2089753221, G2186534006, G2536180224, G3068431579, G3119125940, Novel Mechanisms of Cardiac pH Regulation During Ischemia/Reperfusion, G3708836454, G4165162335, Decoding the enigma of cardiac amplification, G4859167602, G5067448200, A single amino acid in CaV1.2 controls channel activity and arterial tone in hypertension, G5186444726, Topography, regulation and function of PDE- dependent cAMP nanodomains in cardiac myocytes. (Renewal), G618828768, G6947823565, G7057314662, Spatially confined cAMP domains in vascular smooth muscle., Translational Study of Cardiac Dysfunction, G79492955, G8563437507, G8777793247, Training Program in Basic and Translational Cardiovascular Science, cAMP signaling in vascular smooth muscle in health and disease, Multiscale Systems to Elucidate Susceptibility and Cardioprotective Mechanisms in Cardio-oncology, Sympathetic remodeling and ventricular arrhythmia, Functional interactions of cardiac ion channels
OpenAlex ID: W4225517140