Coherency of circadian rhythms in the SCN is governed by the interplay of two coupling factors

Circadian clocks are autonomous oscillators driving daily rhythms in physiology and behavior. In mammals, a network of coupled neurons in the suprachiasmatic nucleus (SCN) is entrained to environmental light-dark cycles and orchestrates the timing of peripheral organs. In each neuron, transcriptional feedbacks generate noisy oscillations. Coupling mediated by neuropeptides such as VIP and AVP lends precision and robustness to circadian rhythms. The detailed coupling mechanisms between SCN neurons are debated. We analyze organotypic SCN slices from neonatal and adult mice in wild-type and multiple knockout conditions. Different degrees of rhythmicity are quantified by pixel-level analysis of bioluminescence data. We use empirical orthogonal functions (EOFs) to characterize spatio-temporal patterns. Simulations of coupled stochastic single cell oscillators can reproduce the diversity of observed patterns. Our combination of data analysis and modeling provides deeper insight into the enormous complexity of the data: (1) Neonatal slices are typically stronger oscillators than adult slices pointing to developmental changes of coupling. (2) Wild-type slices are completely synchronized and exhibit specific spatio-temporal patterns of phases. (3) Some slices of Cry double knockouts obey impaired synchrony that can lead to co–existing rhythms (“splitting”). (4) The loss of VIP-coupling leads to desynchronized rhythms with few residual local clusters. Additional information was extracted from co–culturing slices with rhythmic neonatal wild-type SCNs. These co–culturing experiments were simulated using external forcing terms representing VIP and AVP signaling. The rescue of rhythmicity via co–culturing lead to surprising results, since a cocktail of AVP-antagonists improved synchrony. Our modeling suggests that these counter-intuitive observations are pointing to an antagonistic action of VIP and AVP coupling. Our systematic theoretical and experimental study shows that dual coupling mechanisms can explain the astonishing complexity of spatio-temporal patterns in SCN slices.Author summary: The mammalian circadian clock is orchestrated by a network of coupled neurons. Brain slice preparations allow the analysis of coupling mechanisms mediated by neuropeptides. From bioluminescence recordings, we extract single cell characteristics such as period, amplitude and damping rate. Our data-based stochastic network model involves local coupling between cells and additional external forcing. Available experimental data guide our simulations with two distinct coupling and forcing mechanisms representing the neuropeptides VIP and AVP. We compare our simulations with experiments from neonatal and adult wild-type brain slices and multiple knockouts. Furthermore, we study co–culturing of slices with synchronized neonatal wild-type slices. The extreme complexity of the spatio-temporal patterns is quantified using empirical orthogonal functions (EOFs). The experimental reduction of AVP coupling leads to surprising observations. In double knockouts, inhibition of AVP signaling can improve synchrony, whereas, in triple knockouts, coherency is reduced. Our network modeling shows that these counter-intuitive observations can be explained by an antagonistic action of VIP and AVP signaling. The agreement of experiments and simulations suggests that quite complex spatio-temporal patterns can appear as emergent properties of oscillator networks with dual coupling mechanisms.