Suprachiasmatic Nucleus Network Research Articles

Role of vasoactive intestinal peptide in the light input to the circadian system.

The neuropeptide vasoactive intestinal peptide (VIP) is expressed at high levels in a subset of neurons in the ventral region of the suprachiasmatic nucleus (SCN). While VIP is known to be important for the synchronization of the SCN network, the role of VIP in photic regulation of the circadian system has received less attention. In the present study, we found that the light-evoked increase in electrical activity in vivo was unaltered by the loss of VIP. In the absence of VIP, the ventral SCN still exhibited N-methyl-d-aspartate-evoked responses in a brain slice preparation, although the absolute levels of neural activity before and after treatment were significantly reduced. Next, we used calcium imaging techniques to determine if the loss of VIP altered the calcium influx due to retinohypothalamic tract stimulation. The magnitude of the evoked calcium influx was not reduced in the ventral SCN, but did decline in the dorsal SCN regions. We examined the time course of the photic induction of Period1 in the SCN using insitu hybridization in VIP-mutant mice. We found that the initial induction of Period1 was not reduced by the loss of this signaling peptide. However, the sustained increase in Period1 expression (after 30min) was significantly reduced. Similar results were found by measuring the light induction of cFOS in the SCN. These findings suggest that VIP is critical for longer-term changes within the SCN circuit, but does not play a role in the acute light response.

Cellular Clocks in AVP Neurons of the SCN Are Critical for Interneuronal Coupling Regulating Circadian Behavior Rhythm

The suprachiasmatic nucleus (SCN), the primary circadian pacemaker in mammals, is a network structure composed of multiple types of neurons. Here, we report that mice with a Bmal1 deletion specific to arginine vasopressin (AVP)-producing neurons showed marked lengthening in the free-running period and activity time of behavior rhythms. When exposed to an abrupt 8-hr advance of the light/dark cycle, these mice reentrained faster than control mice did. In these mice, the circadian expression of genes involved in intercellular communications, including Avp, Prokineticin 2, and Rgs16, was drastically reduced in the dorsal SCN, where AVP neurons predominate. In slices, dorsal SCN cells showed attenuated PER2::LUC oscillation with highly variable and lengthened periods. Thus, Bmal1-dependent oscillators of AVP neurons may modulate the coupling of the SCN network, eventually coupling morning and evening behavioral rhythms, by regulating expression of multiple factors important for the network property of these neurons.

Neuromedin S-Producing Neurons Act as Essential Pacemakers in the Suprachiasmatic Nucleus to Couple Clock Neurons and Dictate Circadian Rhythms

Circadian behavior in mammals is orchestrated by neurons within the suprachiasmatic nucleus (SCN), yet the neuronal population necessary for the generation of timekeeping remains unknown. We show that a subset of SCN neurons expressing the neuropeptide neuromedin S (NMS) plays an essential role in the generation of daily rhythms in behavior. We demonstrate that lengthening period within Nms neurons is sufficient to lengthen period of the SCN and behavioral circadian rhythms. Conversely, mice without a functional molecular clock within Nms neurons lack synchronous molecular oscillations and coherent behavioral daily rhythms. Interestingly, we found that mice lacking Nms and its closely related paralog, Nmu, do not lose in vivo circadian rhythms. However, blocking vesicular transmission from Nms neurons with intact cell-autonomous clocks disrupts the timing mechanisms of the SCN, revealing that Nms neurons define a subpopulation of pacemakers that control SCN network synchrony and in vivo circadian rhythms through intercellular synaptic transmission.

Stochastic simulation of the circadian rhythmicity in the SCN neuronal network

The suprachiasmatic nucleus (SCN) of the hypothalamus is the mammalian circadian pacemaker that plays a dominant role in the generation and control of daily physiological and behavioral rhythms. The coupling of the SCN neuronal activity with the solar day–night cycle and the intercellular communication between SCN neurons are the most important factors that ensure the precise 24 h timing of the circadian rhythmicity. In individual cells, however, the timekeeping is not precise because of the inherent randomness in the biochemical reactions. In order to investigate the impact of molecular noise and intercellular connectivity on the circadian network synchronization, we develop a stochastic multicellular model of the SCN. The circadian pacemaker is modeled as a realistic small-world network of neurons and was found to be robust to rather high levels of intrinsic fluctuations. We show that the SCN neuronal network synchronizes well with the photic entraining even at very large levels of stochasticity, whereby intercellular coupling is the required agent that ensures coherent oscillations. Furthermore, we show that the neuronal activity patterns form localized clusters, which are more pronounced in short photoperiods than in long days. The localized synchronization behavior was also found to be affected by the SCN network structure.

Enhanced Phase Resetting in the Synchronized Suprachiasmatic Nucleus Network

The suprachiasmatic nucleus (SCN) adapts to both the external light-dark (LD) cycle and seasonal changes in day length. In short photoperiods, single-cell activity patterns are tightly synchronized (i.e., in phase); in long photoperiods, these patterns are relatively dispersed, causing lower amplitude rhythms. The limit cycle oscillator has been used to describe the SCN's circadian rhythmicity and predicts that following a given perturbation, high-amplitude SCN rhythms will shift less than low-amplitude rhythms. Some studies reported, however, that phase delays are larger when animals are entrained to a short photoperiod. Because phase advances and delays are mediated by partially distinct (i.e., nonoverlapping) biochemical pathways, we investigated the effect of a 4-h phase advance of the LD cycle in mice housed in either short (LD 8:16) or long (LD 16:8) photoperiods. In vitro recordings revealed a significantly larger phase advance in the SCN of mice entrained to short as compared to long photoperiods (4.2 ± 0.3 h v. 1.4 ± 0.9 h, respectively). Surprisingly, in mice with long photoperiods, the behavioral phase shift was larger than the phase shift of the SCN (3.7 ± 0.4 h v. 1.4 ± 0.9 h, respectively). To exclude a confounding influence of running-wheel activity on the magnitude of the shifts of the SCN, we repeated the experiments in the absence of running wheels and found similar shifts in the SCN in vitro in short and long days (3.0 ± 0.5 h v. 0.4 ± 0.9 h, respectively). Interestingly, removal of the running wheel reduced the phase-shifting capacity of mice in long days, leading to similar behavioral shifts in short and long photoperiods (1.0 ± 0.1 h v. 1.0 ± 0.4 h). As the behavioral shifts in the presence of wheels were larger than the shift of the SCN, it is suggested that additional, non-SCN neuronal networks in the brain are involved in regulating the timing of behavioral activity. On the basis of the phase shifts observed in vitro, we conclude that highly synchronized SCN networks with high-amplitude rhythms show a larger phase-shifting capacity than desynchronized networks of low amplitude.

Dynamic Interactions Mediated by Nonredundant Signaling Mechanisms Couple Circadian Clock Neurons

Interactions among suprachiasmatic nucleus (SCN) neurons are required for robust circadian rhythms entrained to local time. To investigate these signaling mechanisms, we developed a functional coupling assay that uniquely captures the dynamic process by which SCN neurons interact. As a population, SCN neurons typically display synchronized rhythms with similar peak times, but will peak 6-12 hr apart after in vivo exposure to long days. Once they are removed from these conditions, SCN neurons resynchronize through a phase-dependent coupling process mediated by both vasoactive intestinal polypeptide (VIP) and GABAA signaling. Notably, GABAA signaling contributes to coupling when the SCN network is in an antiphase configuration, but opposes synchrony under steady-state conditions. Further, VIP acts together with GABAA signaling to couple the network in an antiphase configuration, but promotes synchrony under steady-state conditions by counteracting the actions of GABAA signaling. Thus, SCN neurons interact through nonredundant coupling mechanisms influenced by the state of the network.

Modeling the Seasonal Adaptation of Circadian Clocks by Changes in the Network Structure of the Suprachiasmatic Nucleus

The dynamics of circadian rhythms needs to be adapted to day length changes between summer and winter. It has been observed experimentally, however, that the dynamics of individual neurons of the suprachiasmatic nucleus (SCN) does not change as the seasons change. Rather, the seasonal adaptation of the circadian clock is hypothesized to be a consequence of changes in the intercellular dynamics, which leads to a phase distribution of electrical activity of SCN neurons that is narrower in winter and broader during summer. Yet to understand this complex intercellular dynamics, a more thorough understanding of the impact of the network structure formed by the SCN neurons is needed. To that effect, we propose a mathematical model for the dynamics of the SCN neuronal architecture in which the structure of the network plays a pivotal role. Using our model we show that the fraction of long-range cell-to-cell connections and the seasonal changes in the daily rhythms may be tightly related. In particular, simulations of the proposed mathematical model indicate that the fraction of long-range connections between the cells adjusts the phase distribution and consequently the length of the behavioral activity as follows: dense long-range connections during winter lead to a narrow activity phase, while rare long-range connections during summer lead to a broad activity phase. Our model is also able to account for the experimental observations indicating a larger light-induced phase-shift of the circadian clock during winter, which we show to be a consequence of higher synchronization between neurons. Our model thus provides evidence that the variations in the seasonal dynamics of circadian clocks can in part also be understood and regulated by the plasticity of the SCN network structure.

Central and Peripheral Circadian Clocks in Mammals

The circadian system of mammals is composed of a hierarchy of oscillators that function at the cellular, tissue, and systems levels. A common molecular mechanism underlies the cell-autonomous circadian oscillator throughout the body, yet this clock system is adapted to different functional contexts. In the central suprachiasmatic nucleus (SCN) of the hypothalamus, a coupled population of neuronal circadian oscillators acts as a master pacemaker for the organism to drive rhythms in activity and rest, feeding, body temperature, and hormones. Coupling within the SCN network confers robustness to the SCN pacemaker, which in turn provides stability to the overall temporal architecture of the organism. Throughout the majority of the cells in the body, cell-autonomous circadian clocks are intimately enmeshed within metabolic pathways. Thus, an emerging view for the adaptive significance of circadian clocks is their fundamental role in orchestrating metabolism.

Effect of Network Architecture on Synchronization and Entrainment Properties of the Circadian Oscillations in the Suprachiasmatic Nucleus

In mammals, the suprachiasmatic nucleus (SCN) of the hypothalamus constitutes the central circadian pacemaker. The SCN receives light signals from the retina and controls peripheral circadian clocks (located in the cortex, the pineal gland, the liver, the kidney, the heart, etc.). This hierarchical organization of the circadian system ensures the proper timing of physiological processes. In each SCN neuron, interconnected transcriptional and translational feedback loops enable the circadian expression of the clock genes. Although all the neurons have the same genotype, the oscillations of individual cells are highly heterogeneous in dispersed cell culture: many cells present damped oscillations and the period of the oscillations varies from cell to cell. In addition, the neurotransmitters that ensure the intercellular coupling, and thereby the synchronization of the cellular rhythms, differ between the two main regions of the SCN. In this work, a mathematical model that accounts for this heterogeneous organization of the SCN is presented and used to study the implication of the SCN network topology on synchronization and entrainment properties. The results show that oscillations with larger amplitude can be obtained with scale-free networks, in contrast to random and local connections. Networks with the small-world property such as the scale-free networks used in this work can adapt faster to a delay or advance in the light/dark cycle (jet lag). Interestingly a certain level of cellular heterogeneity is not detrimental to synchronization performances, but on the contrary helps resynchronization after jet lag. When coupling two networks with different topologies that mimic the two regions of the SCN, efficient filtering of pulse-like perturbations in the entrainment pattern is observed. These results suggest that the complex and heterogeneous architecture of the SCN decreases the sensitivity of the network to short entrainment perturbations while, at the same time, improving its adaptation abilities to long term changes.

Quantitative Analysis of Phase Wave of Gene Expression in the Mammalian Central Circadian Clock Network

BackgroundThe suprachiasmatic nucleus (SCN), the master circadian clock, is a heterogeneous oscillator network, yet displays a robust synchronization dynamics. Recent single-cell bioluminescent imaging revealed temporal gradients in circadian clock gene expression in the SCN ex vivo. However, due to technical difficulty in biological approaches to elucidate the entire network structure of the SCN, characteristics of the gradient, which we refer to as phase wave, remain unknown.Methodology/Principal FindingsWe implemented new approaches, i.e., quantitative analysis and model simulation to characterize the phase waves in Per2::Luciferase clock reporter gene expression of the rat SCN slice. Our quantitative study demonstrated not only a high degree of synchronization between the neurons and regular occurrence of the phase wave propagation, but also a significant amount of phase fluctuations contained in the wave. In addition, our simulations based on local coupling model suggest that the intercellular coupling strength estimated by the model simulations is significantly higher than the critical value for generating the phase waves. Model simulations also suggest that heterogeneity of the SCN neurons is one of the main factors causing the phase wave fluctuations. Furthermore, robustness of the SCN network against dynamical noise and variation of the natural frequencies inherent in these neurons was quantitatively assessed.Conclusions/SignificanceTo our knowledge, this is the first quantitative evaluation of the phase wave and further characterization of the SCN neuronal network features generating the wave i.e., intercellular synchrony, phase fluctuation, strong local coupling, heterogeneous periodicity and robustness. Our present study provides an approach, which will lead to a comprehensive understanding of mechanistic and/or biological significance of the phase wave in the central circadian oscillatory system.

How coupling determines the entrainment of circadian clocks

Autonomous circadian clocks drive daily rhythms in physiology and behaviour. A network of coupled neurons, the suprachiasmatic nucleus (SCN), serves as a robust self-sustained circadian pacemaker. Synchronization of this timer to the environmental light-dark cycle is crucial for an organism's fitness. In a recent theoretical and experimental study it was shown that coupling governs the entrainment range of circadian clocks. We apply the theory of coupled oscillators to analyse how diffusive and mean-field coupling affects the entrainment range of interacting cells. Mean-field coupling leads to amplitude expansion of weak oscillators and, as a result, reduces the entrainment range. We also show that coupling determines the rigidity of the synchronized SCN network, i.e. the relaxation rates upon perturbation. %(Floquet exponents). Our simulations and analytical calculations using generic oscillator models help to elucidate how coupling determines the entrainment of the SCN. Our theoretical framework helps to interpret experimental data.

Emergence of Noise-Induced Oscillations in the Central Circadian Pacemaker

Author SummaryThe suprachiasmatic nucleus (SCN) is the master circadian pacemaker in mammals that controls and coordinates physiological processes in a daily manner. The SCN is composed of a network of cells, with each cell acting as an autonomous oscillator. In isolated individual cells, timekeeping is not precise because of the inherent randomness in the biochemical reactions within each cell, involving its core clock components. However, in the SCN network, precise rhythms can emerge because of intercellular coupling. In this article, we study a loss-of-function mutation of BMAL1, a core clock component, which eliminates timekeeping in isolated cells. Surprisingly, in both experiments and mathematical simulations, we find that noisy rhythms emerge from the SCN network even in the presence of this BMAL1 mutation. This random yet coordinated timekeeping has not been observed in previous modeling and experimental work and indicates that a network of cells can utilize noise to help compensate for loss of a physiological function. In normal function, the SCN network mitigates any variability observed in individual cellular rhythms and produces a precise and rhythmic network timekeeping signal. When the individual cells are no longer rhythmic, the coupling pathways within the SCN network can propagate stochastic rhythms that are a reflection of both feed-forward coupling mechanisms and intracellular noise. Thus, in a manner analogous to central pattern generators in neural circuits, rhythmicity can arise as an emergent property of the network in the absence of component pacemaker or oscillator cells.

Neuropeptide‐mediated calcium signaling in the suprachiasmatic nucleus network

Neuroactive peptides and the intracellular calcium concentration ([Ca(2+) ](i) ) play important roles in light-induced modulation of gene expression in the suprachiasmatic nucleus (SCN) neurons that ultimately control behavioral rhythms. Vasoactive intestinal peptide (VIP) and arginine vasopressin (AVP) are expressed rhythmically within populations of SCN neurons. Pituitary adenylate cyclase-activating peptide (PACAP) is released from retinohypothalamic tract (RHT) terminals synapsing on SCN neurons. Nociceptin/orphanin FQ (OFQ) receptors are functionally expressed in the SCN. We examined the role of several neuropeptides on Ca(2+) signaling, simultaneously imaging multiple neurons within the SCN neural network. VIP reduced the [Ca(2+) ](i) in populations of SCN neurons during the day, but had little effect at night. Stimulation of the RHT at frequencies that simulate light input signaling evoked transient [Ca(2+) ](i) elevations that were not altered by VIP. AVP elevated the [Ca(2+) ](i) during both the day and night, PACAP produced variable responses, and OFQ induced a reduction in the [Ca(2+) ](i) similar to VIP. During the day, VIP lowered the [Ca(2+) ](i) to near nighttime levels, while AVP elevated [Ca(2+) ](i) during both the day and night, suggesting that the VIP effects on [Ca(2+) ](i) were dependent, and the AVP effects independent of the action potential firing activity state of the neuron. We hypothesize that VIP and AVP regulate, at least in part, Ca(2+) homeostasis in SCN neurons and may be a major point of regulation for SCN neuronal synchronization.

Jet lag syndrome: circadian organization, pathophysiology, and management strategies

The circadian system regulates the cyclical occurrence of wakefulness and sleep through a series of oscillatory networks that comprise two different theoretical processes. The suprachiasmatic nucleus (SCN) of the hypothalamus contains the master oscillatory network necessary for coordinating these daily rhythms, and in addition to its ability to robustly generate rhythms, it can also synchronize to environmental light cues. During jet lag, abrupt shifts in the environmental light–dark cycle temporarily desynchronize the SCN and downstream oscillatory networks from each other, resulting in increased sleepiness and impaired daytime functioning. Polysomnographic data show that not only does jet lag result in changes of sleep–wake timing, but also in different aspects of sleep architecture. This type of circadian misalignment can further lead to a cluster of symptoms including major metabolic, cardiovascular, psychiatric, and neurological impairments. There are a number of treatment options for jet lag involving bright light exposure, melatonin, and use of hypnotics, but their efficacy greatly depends on their time of use, the length of time in the new time zone, and the specific circadian disturbance involved. The aim of this review is to provide mechanistic links between the fields of sleep and circadian rhythms to understand the biological basis of jet lag and to apply this information to clinical management strategies.

Suprachiasmatic nucleus: cell autonomy and network properties.

The suprachiasmatic nucleus (SCN) is the primary circadian pacemaker in mammals. Individual SCN neurons in dispersed culture can generate independent circadian oscillations of clock gene expression and neuronal firing. However, SCN rhythmicity depends on sufficient membrane depolarization and levels of intracellular calcium and cAMP. In the intact SCN, cellular oscillations are synchronized and reinforced by rhythmic synaptic input from other cells, resulting in a reproducible topographic pattern of distinct phases and amplitudes specified by SCN circuit organization. The SCN network synchronizes its component cellular oscillators, reinforces their oscillations, responds to light input by altering their phase distribution, increases their robustness to genetic perturbations, and enhances their precision. Thus, even though individual SCN neurons can be cell-autonomous circadian oscillators, neuronal network properties are integral to normal function of the SCN.

Reorganization of Suprachiasmatic Nucleus Networks under 24-h LDLD Conditions

The suprachiasmatic nucleus (SCN), locus of the master circadian clock in the brain, is comprised of multioscillator neural networks that are highly plastic in responding to environmental lighting conditions. Under a 24-h light:dark:light:dark (LDLD) cycle, hamsters bifurcate their circadian locomotor activity such that wheel running occurs in each of the 2 daily dark periods with complete inactivity in between. In the present study, we explored the neural underpinning of this behavioral bifurcation. Using calbindin (CalB)- containing cells of the SCN as a regional marker, we characterized PER1 and c-FOS expression in the core and shell SCN subregions. In LD-housed animals, it is known that PER1 and c-FOS in the core and shell region are in phase with each other. In contrast, in behaviorally bifurcated animals housed in LDLD, the core and shell SCN exhibit antiphase rhythms of PER1. Furthermore, cells in the core show high FOS expression in each photophase of the LDLD cycle. The activation of FOS in the core is light driven and disappears rapidly when the photophase is replaced by darkness. The results suggest that bifurcated activity bouts in daytime and nighttime are associated with oscillating groups of cells in the core and shell subregions, respectively, and support the notion that reorganization of SCN networks underlies changes in behavioral responses under different environmental lighting conditions.

GABAergic signaling induces divergent neuronal Ca2+ responses in the suprachiasmatic nucleus network

Intercellular communication between gamma-aminobutyric acid (GABA)ergic suprachiasmatic nucleus (SCN) neurons facilitates light-induced phase changes and synchronization of individual neural oscillators within the SCN network. We used ratiometric Ca(2+) imaging techniques to record changes in the intracellular calcium concentration ([Ca(2+)](i)) to study the role of GABA in interneuronal communication and the response of the SCN neuronal network to optic nerve stimulations that mimic entraining light signals. Stimulation of the retinohypothalamic tract (RHT) evoked divergent Ca(2+) responses in neurons that varied regionally within the SCN with a pattern that correlated with those evoked by pharmacological GABA applications. GABA(A) and GABA(B) receptor agonists and antagonists were used to evaluate components of the GABA-induced changes in [Ca(2+)](i). Application of the GABA(A) receptor antagonist gabazine induced changes in baseline [Ca(2+)](i) in a direction opposite to that evoked by GABA, and similarly altered the RHT stimulation-induced Ca(2+) response. GABA application induced Ca(2+) responses varied in time and region within the SCN network. The NKCC1 cotransporter blocker, bumetanide, and L-type calcium channel blocker, nimodipine, attenuated the GABA-induced rise of [Ca(2+)](i). These results suggest that physiological GABA induces opposing effects on [Ca(2+)](i) based on the chloride equilibrium potential, and may play an important role in neuronal Ca(2+) balance, synchronization and modulation of light input signaling in the SCN network.

Limits of single-cell autonomy in the suprachiasmatic nucleus

The suprachiasmatic nucleus (SCN) is the master circadian pacemaker in the mammalian brain, responsible for coordinating circadian (ca 24 h) rhythms throughout the body. When individual SCN neurons are dispersed in low density culture, they generate independent circadian oscillations in neuronal firing rate. This observation has led to the widely accepted conclusion that single SCN neurons are autonomous circadian oscillators. More recent studies using bioluminescence imaging to monitor rhythms of clock gene expression have indicated that fibroblasts, too, are autonomous circadian oscillators. But does this mean that the fibroblast should “replace the SCN as the in vitro model of choice”? On the contrary, the SCN is much more than a population of independent cellular oscillators. SCN function depends on important network interactions that allow for cell synchrony, enhanced pacemaker precision, reinforcement of rhythm amplitude, and robustness against genetic perturbations. The distribution of phases of cells within the SCN network also permits variously phased rhythmic output signals, as well as the encoding of seasonal changes in photoperiod and other features of environmental lighting history. Thus, unlike fibroblasts, SCN neurons are designed to be “team players,” and SCN network interactions are integral to the normal function of the SCN as a circadian pacemaker.

Expression of clock genes in the suprachiasmatic nucleus: Effect of environmental lighting conditions

The suprachiasmatic nucleus (SCN) is the anatomical substrate for the principal circadian clock coordinating daily rhythms in a vast array of behavioral and physiological responses. Individual SCN neurons are cellular oscillators and are organized into a multi-oscillator network following unique spatiotemporal patterns. The rhythms generated in the SCN are generally entrained to the environmental light dark cycle, which is the most salient cue influencing the network organization of the SCN. The neural network in the SCN is a heterogeneous structure, containing two major compartments identified by applying physiological and functional criteria, namely the retinorecipient core region and the highly rhythmic shell region. Changes in the environmental lighting condition are first detected and processed by the core region, and then conveyed to the rest of the SCN, leading to adaptive responses of the entire network. This review will focus on the studies that explore the responses of the SCN network by examining the expression of clock genes, under various lighting paradigms, such as acute light exposure, lighting schedules or exposure to different light durations. The results will be discussed under the framework of functionally distinct SCN sub regions and oscillator groups. The evidence presented here suggests that the environmental lighting conditions alter the spatiotemporal organization of the cellular oscillators within the SCN, which consequently affect the overt rhythms in behavior and physiology. Thus, information on how the SCN network elements respond to environmental cues is key to understanding the human health problems that stem from circadian rhythm disruption.

Light-Dark Cycle Memory in the Mammalian Suprachiasmatic Nucleus

The mammalian circadian oscillator, or suprachiasmatic nucleus (SCN), contains several thousand clock neurons in its ventrolateral division, many of which are spontaneous oscillators with period lengths that range from 22 to 28 h. In complete darkness, this network synchronizes through the exchange of action potentials that release vasoactive intestinal polypeptide, striking a compromise, free-running period close to 24 h long. We entrained Siberian hamsters to various light-dark cycles and then tracked their activity into constant darkness to show that they retain a memory of the previous light-dark cycle before returning to their own free-running period. Employing Leloup-Goldbeter mammalian clock neurons we model the ventrolateral SCN network and show that light acting weakly upon a strongly rhythmic vasoactive intestinal polypeptide oscillation can explain the observed light-dark cycle memory. In addition, light is known to initiate a mitogen-activated protein kinase signaling cascade that induces transcription of both per and mkp1 phosphatase. We show that the ensuing phosphatase-kinase interaction can account for the dead zone in the mammalian phase response curve and hypothesize that the SCN behaves like a lock-in amplifier to entrain to the light edges of the circadian day.