Suprachiasmatic Nucleus Network Research Articles

Neuron-Astrocyte Interactions and Circadian Timekeeping in Mammals.

Almost every facet of our behavior and physiology varies predictably over the course of day and night, anticipating and adapting us to their associated opportunities and challenges. These rhythms are driven by endogenous biological clocks that, when deprived of environmental cues, can continue to oscillate within a period of approximately 1 day, hence circa-dian. Normally, retinal signals synchronize them to the cycle of light and darkness, but disruption of circadian organization, a common feature of modern lifestyles, carries considerable costs to health. Circadian timekeeping pivots around a cell-autonomous molecular clock, widely expressed across tissues. These cellular timers are in turn synchronized by the principal circadian clock of the brain: the hypothalamic suprachiasmatic nucleus (SCN). Intercellular signals make the SCN network a very powerful pacemaker. Previously, neurons were considered the sole SCN timekeepers, with glial cells playing supportive roles. New discoveries have revealed, however, that astrocytes are active partners in SCN network timekeeping, with their cell-autonomous clock regulating extracellular glutamate and GABA concentrations to control circadian cycles of SCN neuronal activity. Here, we introduce circadian timekeeping at the cellular and SCN network levels before focusing on the contributions of astrocytes and their mutual interaction with neurons in circadian control in the brain.

The Suprachiasmatic Nucleus at 50: Looking Back, Then Looking Forward.

It has been 50 years since the suprachiasmatic nucleus (SCN) was first identified as the central circadian clock and 25 years since the last overview of developments in the field was published in the Journal of Biological Rhythms. Here, we explore new mechanisms and concepts that have emerged in the subsequent 25 years. Since 1997, methodological developments, such as luminescent and fluorescent reporter techniques, have revealed intricate relationships between cellular and network-level mechanisms. In particular, specific neuropeptides such as arginine vasopressin, vasoactive intestinal peptide, and gastrin-releasing peptide have been identified as key players in the synchronization of cellular circadian rhythms within the SCN. The discovery of multiple oscillators governing behavioral and physiological rhythms has significantly advanced our understanding of the circadian clock. The interaction between neurons and glial cells has been found to play a crucial role in regulating these circadian rhythms within the SCN. Furthermore, the properties of the SCN network vary across ontogenetic stages. The application of cell type-specific genetic manipulations has revealed components of the functional input-output system of the SCN and their correlation with physiological functions. This review concludes with the high-risk effort of identifying open questions and challenges that lie ahead.

Expression of the vesicular GABA transporter within neuromedin S+ neurons sustains behavioral circadian rhythms.

The suprachiasmatic nucleus (SCN) of the hypothalamus is the site of a central circadian clock that orchestrates overt rhythms of physiology and behavior. Circadian timekeeping requires intercellular communication among SCN neurons, and multiple signaling pathways contribute to SCN network coupling. Gamma-aminobutyric acid (GABA) is produced by virtually all SCN neurons, and previous work demonstrates that this transmitter regulates coupling in the adult SCN but is not essential for the nucleus to sustain overt circadian rhythms. Here, we show that the deletion of the gene that codes for the GABA vesicular transporter Vgat from neuromedin-S (NMS)+ neurons-a subset of neurons critical for SCN function-causes arrhythmia of locomotor activity and sleep. Further, NMS-Vgat deletion impairs intrinsic clock gene rhythms in SCN explants cultured ex vivo. Although vasoactive intestinal polypeptide (VIP) is critical for SCN function, Vgat deletion from VIP-expressing neurons did not lead to circadian arrhythmia in locomotor activity rhythms. Likewise, adult SCN-specific deletion of Vgat led to mild impairment of behavioral rhythms. Our results suggest that while the removal of GABA release from the adult SCN does not affect the pacemaker's ability to sustain overt circadian rhythms, its removal from a critical subset of neurons within the SCN throughout development removes the nucleus ability to sustain circadian rhythms. Our findings support a model in which SCN GABA release is critical for the developmental establishment of intercellular network properties that define the SCN as a central pacemaker.

The effects of long-range connections on navigation in suprachiasmatic nucleus networks

The effects of long-range connections on navigation in suprachiasmatic nucleus networks

Vasculature of the Suprachiasmatic Nucleus: Pathways for Diffusible Output Signals.

Transplant studies demonstrate unequivocally that the suprachiasmatic nucleus (SCN) produces diffusible signals that can sustain circadian locomotor rhythms. There is a vascular portal pathway between the SCN and the organum vasculosum of the lamina terminalis in mouse brain. Portal pathways enable low concentrations of neurosecretions to reach specialized local targets without dilution in the systemic circulation. To explore the SCN vasculature and the capillary vessels whereby SCN neurosecretions might reach portal vessels, we investigated the blood vessels (BVs) of the core and shell SCN. The arterial supply of the SCN differs among animals, and in some animals, there are differences between the 2 sides. The rostral SCN is supplied by branches from either the superior hypophyseal artery (SHpA) or the anterior cerebral artery or the anterior communicating artery. The caudal SCN is consistently supplied by the SHpA. The rostral SCN is drained by the preoptic vein, while the caudal is drained by the basal vein, with variations in laterality of draining vessels. In addition, several key features of the core and shell SCN regions differ: Median BV diameter is significantly smaller in the shell than the core based on confocal image measurements, and a similar trend occurs in iDISCO-cleared tissue. In the cleared tissue, whole BV length density and surface area density are significantly greater in the shell than the core. Finally, capillary length density is also greater in the shell than the core. The results suggest three hypotheses: First, the distinct arterial and venous systems of the rostral and caudal SCN may contribute to the in vivo variations of metabolic and neural activities observed in SCN networks. Second, the dense capillaries of the SCN shell are well positioned to transport blood-borne signals. Finally, variations in SCN vascular supply and drainage may contribute to inter-animal differences.

Free running period affected by network structures of suprachiasmatic nucleus neurons exposed to constant light

Exposed to the natural light-dark cycle, 24 h rhythms exist in behavioral and physiological processes of living beings. Interestingly, under constant darkness or constant light, living beings can maintain a robust endogenous rhythm with a free running period (FRP) close to 24 h. In mammals, the circadian rhythm is coordinated by a master clock located in the suprachiasmatic nucleus (SCN) of the brain, which is composed of about twenty thousand self-oscillating neurons. These SCN neurons form a heterogenous network to output a robust rhythm. Thus far, the exact network topology of the SCN neurons is unknown. In this article, we examine the effect of the SCN network structure on the FRP when exposed to constant light by a Poincaré model. Four typical network structures are considered, including a nearest-neighbor coupled network, a Newman–Watts small world network, an ErdÖs–Rényi random network and a Barabási–Albert (BA) scale free network. The results show that the FRP is longest in the BA network, because the BA network is characterized by the most heterogeneous structure among these four types of networks. These findings are not affected by the average node degree of the SCN network or the value of relaxation rate of the SCN neuronal oscillators. Our findings contribute to the understanding of how the network structure of the SCN neurons influences the FRP.

Aging affects GABAergic function and calcium homeostasis in the mammalian central clock.

Aging impairs the function of the central circadian clock in mammals, the suprachiasmatic nucleus (SCN), leading to a reduction in the output signal. The weaker timing signal from the SCN results in a decline in rhythm strength in many physiological functions, including sleep-wake patterns. Accumulating evidence suggests that the reduced amplitude of the SCN signal is caused by a decreased synchrony among the SCN neurons. The present study was aimed to investigate the hypothesis that the excitation/inhibition (E/I) balance plays a role in synchronization within the network. Using calcium (Ca2+) imaging, the polarity of Ca2+ transients in response to GABA stimulation in SCN slices of old mice (20-24 months) and young controls was studied. We found that the amount of GABAergic excitation was increased, and that concordantly the E/I balance was higher in SCN slices of old mice when compared to young controls. Moreover, we showed an effect of aging on the baseline intracellular Ca2+ concentration, with higher Ca2+ levels in SCN neurons of old mice, indicating an alteration in Ca2+ homeostasis in the aged SCN. We conclude that the change in GABAergic function, and possibly the Ca2+ homeostasis, in SCN neurons may contribute to the altered synchrony within the aged SCN network.

Somatostatin regulates central clock function and circadian responses to light

Daily and annual changes in light are processed by central clock circuits that control the timing of behavior and physiology. The suprachiasmatic nucleus (SCN) in the anterior hypothalamus processes daily photic inputs and encodes changes in day length (i.e., photoperiod), but the SCN circuits that regulate circadian and photoperiodic responses to light remain unclear. Somatostatin (SST) expression in the hypothalamus is modulated by photoperiod, but the role of SST in SCN responses to light has not been examined. Our results indicate that SST signaling regulates daily rhythms in behavior and SCN function in a manner influenced by sex. First, we use cell-fate mapping to provide evidence that SST in the SCN is regulated by light viade novoSst activation. Next, we demonstrate that Sst -/-mice display enhanced circadian responses to light, with increased behavioral plasticity to photoperiod, jetlag, and constant light conditions. Notably, lack of Sst -/-eliminated sex differences in photic responses due to increased plasticity in males, suggesting that SST interacts with clock circuits that process light differently in each sex. Sst -/-mice also displayed an increase in the number of retinorecipient neurons in the SCN core, which express a type of SST receptor capable of resetting the molecular clock. Last, we show that lack of SST signaling modulates central clock function by influencing SCN photoperiodic encoding, network after-effects, and intercellular synchrony in a sex-specific manner. Collectively, these results provide insight into peptide signaling mechanisms that regulate central clock function and its response to light.

A Mean-Field Firing-Rate Model for the Suprachiasmatic Nucleus

We present a mean-field formalism for modeling firing-rate statistics of brain regions whose neurons exhibit atypical firing patterns and heterogeneous electrophysiological properties. We apply the formalism to the suprachiasmatic nucleus (SCN)—the human circadian pacemaker—whose neurons can intrinsically exhibit depolarized low-amplitude membrane oscillations (DLAMOs), depolarization block (DB), and standard action potential firing at different times of day. Further, gamma-aminobutyric acid reversal potentials and molecular circadian phases of SCN neurons, among other properties, vary across the network and/or slowly over time. Our formalism consists of a system of integro-differential equations describing the time evolution of the mean and standard deviation of synaptic conductances across the network. Electrophysiological properties of SCN neurons are incorporated by computing responses to synaptic conductance inputs of a Hodgkin–Huxley-type SCN neuron model that exhibits DLAMOs and DB. Such responses are then averaged over distributions of relevant quantities and included in the differential equations. Results suggest mechanisms by which physiologically relevant changes to firing activities may arise, highlighting means by which the amplitude of firing rates may shrink, the standard deviation of firing rates may grow, and by which a mid-day dip in firing rates may appear. For instance, results show that a large spread in circadian phases across SCN neurons reduces the size of oscillations in SCN network firing activity across the 24-hour day, identifying a mechanism by which heterogeneities in neuron electrophysiology could influence circadian rhythms.

Circadian rhythmogenesis from neuromodulator- and molecular clock-mediated network mixed feedback*

The suprachiasmatic nucleus (SCN) is a subset of neurons in charge of timekeeping circadian phenomena. As a network, it is capable of maintaining periodic activity with stable phase-locking patterns in spite of heterogeneous nodes. The mechanisms underlying SCN rhythmogenesis are still largely debated. We propose a novel biophysical SCN network model in which circadian rhythmicity emerges from the interaction of neuromodulator-mediated network positive feedback and molecular clock-mediated cellular negative feedback. Because neuromodulator action is much faster than molecular clock dynamics, our model reveals that the same “mixed” (i.e., fast positive, slow negative) feedback governing neuronal excitability also rules circadian oscillations but on several orders of magnitude slower timescales. It also reveals that, when dynamics are sufficiently slow, neuromodulators can create, instead of solely modulate, feedback loops. A mathematical abstraction of the derived SCN network makes novel predictions about the action of various SCN neuromodulators.

Light-induced synchronization of the SCN coupled oscillators and implications for entraining the HPA axis.

The suprachiasmatic nucleus (SCN) synchronizes the physiological rhythms to the external light-dark cycle and tunes the dynamics of circadian rhythms to photoperiod fluctuations. Changes in the neuronal network topologies are suggested to cause adaptation of the SCN in different photoperiods, resulting in the broader phase distribution of neuron activities in long photoperiods (LP) compared to short photoperiods (SP). Regulated by the SCN output, the level of glucocorticoids is elevated in short photoperiod, which is associated with peak disease incidence. The underlying coupling mechanisms of the SCN and the interplay between the SCN and the HPA axis have yet to be fully elucidated. In this work, we propose a mathematical model including a multiple-cellular SCN compartment and the HPA axis to investigate the properties of the circadian timing system under photoperiod changes. Our model predicts that the probability-dependent network is more energy-efficient than the distance-dependent network. Coupling the SCN network by intra-subpopulation and inter-subpopulation forces, we identified the negative correlation between robustness and plasticity of the oscillatory network. The HPA rhythms were predicted to be strongly entrained to the SCN rhythms with a pro-inflammatory high-amplitude glucocorticoid profile under SP. The fast temporal topology switch of the SCN network was predicted to enhance synchronization when the synchronization is not complete. These synchronization and circadian dynamics alterations might govern the seasonal variation of disease incidence and its symptom severity.

Poincaré model shows how heterogeneity in light sensitivity can alter circadian clock function

Exposed to the natural light-dark cycle, living beings show robust 24 h rhythms in physiology and behavior. Interestingly, even in the absence of a light-dark cycle, for example in constant conditions, such as under the constant darkness or the constant light, living beings maintain a robust rhythm of which the endogenous period (named free running period, FRP) is close to 24 h. The endogenous rhythms are regulated by a master clock located in the suprachiasmatic nucleus (SCN) of mammals, where the SCN neurons show heterogeneity in the sensitivity to the light. In this article, we examined how this heterogeneity influences the FRP under constant light. Using a Poincaré model for the SCN network it is shown that the FRP increases with the increase of the degree of heterogeneity in the sensitivity of neuronal subpopulations to light. Moreover, the presence of a critical value where the periods of the subpopulation diverge, presents a mechanism dictating how some animals remain rhythmic under constant light conditions, while others lose their rhythms completely. Our findings help to understand how the neuronal heterogeneity to light sensitivity in the SCN influences the circadian behavior of the animal.

Network rewiring and plasticity promotes synchronization of suprachiasmatic nucleus neurons.

In mammals, circadian rhythms throughout the body are orchestrated by the master clock in the hypothalamic suprachiasmatic nucleus (SCN), where SCN neurons are coupled with neurotransmitters to generate a uniform circadian rhythm. How the SCN circadian rhythm is so robust and flexible is, however, unclear. In this paper, we propose a temporal SCN network model and investigate the effects of dynamical rewiring and flexible coupling due to synaptic plasticity on the synchronization of the neural network in SCN. In networks consisting of simple Poincaré oscillators and complex circadian clocks, we found that dynamical rewiring and coupling plasticity enhance the synchronization in inhomogeneous networks. We verified the effect of enhanced synchronization in different architectures of random, scale-free, and small-world networks. A simple mean-field analysis for synchronization in plastic networks is proposed. Intuitively, the synchronization is greatly enhanced because both the random rewiring and coupling plasticity in the heterogeneous network have effectively increased the coupling strength in the whole network. Our results suggest that a proper network model for the master SCN circadian rhythm needs to take into account the effects of dynamical changes in topology and plasticity in neuron interactions that could help the brain to generate a robust circadian rhythm for the whole body.

Optogenetic stimulation of VIPergic SCN neurons induces photoperiodic-like changes in the mammalian circadian clock.

Circadian clocks play key roles in how organisms respond to and even anticipate seasonal change in day length, or photoperiod. In mammals, photoperiod is encoded by the central circadian pacemaker in the brain, the suprachiasmatic nucleus (SCN). The subpopulation of SCN neurons that secrete the neuropeptide VIP mediates the transmission of light information within the SCN neural network, suggesting a role for these neurons in circadian plasticity in response to light information that has yet to be directly tested. Here, we used in vivo optogenetic stimulation of VIPergic SCN neurons followed by ex vivo PERIOD 2::LUCIFERASE (PER2::LUC) bioluminescent imaging to test whether activation of this SCN neuron subpopulation can induce SCN network changes that are hallmarks of photoperiodic encoding. We found that optogenetic stimulation designed to mimic a long photoperiod indeed altered subsequent SCN entrained phase, increased the phase dispersal of PER2 rhythms within the SCN network, and shortened SCN free-running period-similar to the effects of a true extension of photoperiod. Optogenetic stimulation also induced analogous changes on related aspects of locomotor behaviour in vivo. Thus, selective activation of VIPergic SCN neurons induces photoperiodic network plasticity in the SCN that underpins photoperiodic entrainment of behaviour.

Network Structure of the Master Clock Is Important for Its Primary Function.

A master clock located in the suprachiasmatic nucleus (SCN) regulates the circadian rhythm of physiological and behavioral activities in mammals. The SCN has two main functions in the regulation: an endogenous clock produces the endogenous rhythmic signal in body rhythms, and a calibrator synchronizes the body rhythms to the external light-dark cycle. These two functions have been determined to depend on either the dynamic behaviors of individual neurons or the whole SCN neuronal network. In this review, we first introduce possible network structures for the SCN, as revealed by time series analysis from real experimental data. It was found that the SCN network is heterogeneous and sparse, that is, the average shortest path length is very short, some nodes are hubs with large node degrees but most nodes have small node degrees, and the average node degree of the network is small. Secondly, the effects of the SCN network structure on the SCN function are reviewed based on mathematical models of the SCN network. It was found that robust rhythms with large amplitudes, a high synchronization between SCN neurons and a large entrainment ability exists mainly in small-world and scale-free type networks, but not other types. We conclude that the SCN most probably is an efficient small-world type or scale-free type network, which drives SCN function.

Roles of Neuropeptides, VIP and AVP, in the Mammalian Central Circadian Clock.

In mammals, the central circadian clock is located in the suprachiasmatic nucleus (SCN) of the hypothalamus. Individual SCN cells exhibit intrinsic oscillations, and their circadian period and robustness are different cell by cell in the absence of cellular coupling, indicating that cellular coupling is important for coherent circadian rhythms in the SCN. Several neuropeptides such as arginine vasopressin (AVP) and vasoactive intestinal polypeptide (VIP) are expressed in the SCN, where these neuropeptides function as synchronizers and are important for entrainment to environmental light and for determining the circadian period. These neuropeptides are also related to developmental changes of the circadian system of the SCN. Transcription factors are required for the formation of neuropeptide-related neuronal networks. Although VIP is critical for synchrony of circadian rhythms in the neonatal SCN, it is not required for synchrony in the embryonic SCN. During postnatal development, the clock genes cryptochrome (Cry)1 and Cry2 are involved in the maturation of cellular networks, and AVP is involved in SCN networks. This mini-review focuses on the functional roles of neuropeptides in the SCN based on recent findings in the literature.

GABA from vasopressin neurons regulates the time at which suprachiasmatic nucleus molecular clocks enable circadian behavior

The suprachiasmatic nucleus (SCN), the central circadian pacemaker in mammals, is a network structure composed of multiple types of γ-aminobutyric acid (GABA)-ergic neurons and glial cells. However, the roles of GABA-mediated signaling in the SCN network remain controversial. Here, we report noticeable impairment of the circadian rhythm in mice with a specific deletion of the vesicular GABA transporter in arginine vasopressin (AVP)-producing neurons. These mice showed disturbed diurnal rhythms of GABAA receptor-mediated synaptic transmission in SCN neurons and marked lengthening of the activity time in circadian behavioral rhythms due to the extended interval between morning and evening locomotor activities. Synchrony of molecular circadian oscillations among SCN neurons did not significantly change, whereas the phase relationships between SCN molecular clocks and circadian morning/evening locomotor activities were altered significantly, as revealed by PER2::LUC imaging of SCN explants and in vivo recording of intracellular Ca2+ in SCN AVP neurons. In contrast, daily neuronal activity in SCN neurons in vivo clearly showed a bimodal pattern that correlated with dissociated morning/evening locomotor activities. Therefore, GABAergic transmission from AVP neurons regulates the timing of SCN neuronal firing to temporally restrict circadian behavior to appropriate time windows in SCN molecular clocks.

Neural Mechanisms Underlying the Central Circadian Clock of the Suprachiasmatic Nucleus

Circadian rhythms are oscillations with an approximately 24-h period and appear in most of the physiological events of our body. The suprachiasmatic nucleus (SCN) of the hypothalamus functions as the central circadian clock in mammals and entrains to the environmental light/dark (day/night) cycle. Here, I briefly review the molecular, cellular, and anatomical structures of the SCN, present findings of recent studies on the differential roles of multiple neuropeptides and neuropeptide-expressing neurons in the SCN, and discuss the mechanisms of the SCN network.

Brief light exposure at dawn and dusk can encode day-length in the neuronal network of the mammalian circadian pacemaker.

The central circadian pacemaker in mammals, the suprachiasmatic nucleus (SCN), is important for daily as well as seasonal rhythms. The SCN encodes seasonal changes in day length by adjusting phase distribution among oscillating neurons thereby shaping the output signal used for adaptation of physiology and behavior. It is well-established that brief light exposure at the beginning and end of the day, also referred to as "skeleton" light pulses, are sufficient to evoke the seasonal behavioral phenotype. However, the effect of skeleton light exposure on SCN network reorganization remains unknown. Therefore, we exposed mice to brief morning and evening light pulses that mark the time of dawn and dusk in a short winter- or a long summer day. Single-cell PER2::LUC recordings, electrophysiological recordings of SCN activity, and measurements of GABA response polarity revealed that skeleton light-regimes affected the SCN network to the same degree as full photoperiod. These results indicate the powerful, yet potentially harmful effects of even relatively short light exposures during the evening or night for nocturnal animals.

Seasonal plasticity in GABAA signaling is necessary for restoring phase synchrony in the master circadian clock network.

Annual changes in the environment threaten survival, and numerous biological processes in mammals adjust to this challenge via seasonal encoding by the suprachiasmatic nucleus (SCN). To tune behavior according to day length, SCN neurons display unified rhythms with synchronous phasing when days are short, but will divide into two sub-clusters when days are long. The transition between SCN states is critical for maintaining behavioral responses to seasonal change, but the mechanisms regulating this form of neuroplasticity remain unclear. Here we identify that a switch in chloride transport and GABAA signaling is critical for maintaining state plasticity in the SCN network. Further, we reveal that blocking excitatory GABAA signaling locks the SCN into its long day state. Collectively, these data demonstrate that plasticity in GABAA signaling dictates how clock neurons interact to maintain environmental encoding. Further, this work highlights factors that may influence susceptibility to seasonal disorders in humans.