Deciphering cellular signals in adult mouse sinoatrial node cells

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

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Article number103693
JournaliScience
Volume25
Issue number1
DOIs
StatePublished - Jan 21 2022

Keywords

  • Biology experimental methods
  • Cell biology
  • Functional aspects of cell biology

ASJC Scopus subject areas

  • General

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