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Deciphering cellular signals in adult mouse sinoatrial node cells.

iScience (2022-01-18)
Gopireddy R Reddy, Lu Ren, Phung N Thai, Jessica L Caldwell, Manuela Zaccolo, Julie Bossuyt, Crystal M Ripplinger, Yang K Xiang, Madeline Nieves-Cintrón, Nipavan Chiamvimonvat, Manuel F Navedo
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, 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.

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