Deciphering Cellular Signals in Adult Mouse Sinoatrial Node Cells

The flexibility of manipulating the mouse genome makes cells from this species an ideal model to study cellular signals that control organ function. Genetically-modified mice provide opportunities decipher how pacemaking sinoatrial node (SAN) are regulated by several second messengers and signaling molecules. However, detailed information on spatiotemporal dynamics these events in adult SAN has been limited due challenge visualizing them live cells. Here, a method is described for culturing immediate infection with genetically-encoded FRET-based biosensors examine events. maintained culture media containing blebbistatin or (S)-nitro-blebbistatin retain their elongated morphology action potential (AP) waveform up 40 hours. condition does not change β-adrenergic-mediated cAMP signal as determined freshly dissociated cultured cardiac-specific reporter mouse. expressing sensors different compartments show distinct pools. Examination cyclic GMP (cGMP), protein kinase A (PKA), Ca2+/CaM II (CaMKII) D (PKD) activity specific FRET also unique responses stimuli Isolation heart failure reveal decrease cGMP signaling. Thus, reliable efficient developed facilitate studies network during physiological pathological conditions.