Molecular basis of Period 1 regulation by adrenergic signaling in the heart

The cardiac circadian clock is responsible for the modulation of different myocardial processes, and its dysregulation has been linked to disease development. How this clock machinery is regulated in the heart remains an open question. Because noradrenaline (NE) can act as a zeitgeber in cardiomyocytes, we tested the hypothesis that adrenergic signaling resets cardiac clock gene expression in vivo. In its anti-phase with Clock and Bmal1, cardiac Per1 abundance increased during the dark phase, concurrent with the rise in heart rate and preceded by an increase in NE levels. Sympathetic denervation altered Bmal1 and Clock amplitude, while Per1 was affected in both amplitude and oscillatory pattern. We next treated mice with a beta-adrenergic receptor (beta-AR) blocker. Strikingly, the beta-AR blockade during the day suppressed the nocturnal increase in Per1 mRNA, without altering Clock or Bmal1. In contrast, activating beta-AR with isoproterenol (ISO) promoted an increase in Per1 expression, demonstrating its responsiveness to adrenergic input. Inhibitors of ERK1/2 and CREB attenuated ISO-induced Per1 expression. Upstream of ERK1/2, PI3K gamma mediated ISO induction of Per1 transcription, while activation of beta 2-AR, but not beta 1-AR induced increases in ERK1/2 phosphorylation and Per1 expression. Consistent with the beta 2-induction of Per1 mRNA, ISO failed to activate ERK1/2 and elevate Per1 in the heart of beta 2-AR(-/-) mice, whereas a beta 2-AR antagonist attenuated the nocturnal rise in Per1 expression. Our study established a link between NE/beta 2-AR signaling and Per1 oscillation via the PI3Ky-ERK1/2-CREB pathway, providing a new framework for understanding the physiological mechanism involved in resetting cardiac clock genes.