Photic Entrainment Of Circadian Rhythms Research Articles

Vasopressin casts light on the suprachiasmatic nucleus.

A subpopulation of retinal ganglion cells expresses the neuropeptide vasopressin. These retinal ganglion cells project predominately to our biological clock, the suprachiasmatic nucleus (SCN). Light-induced vasopressin release enhances the responses of SCN neurons to light. It also enhances expression of genes involved in photo-entrainment of biological rhythms. In all animals, the transition between night and day engages a host of physiological and behavioural rhythms. These rhythms depend not on the rods and cones of the retina, but on retinal ganglion cells (RGCs) that detect the ambient light level in the environment. These project to the suprachiasmatic nucleus (SCN) of the hypothalamus to entrain circadian rhythms that are generated within the SCN. The neuropeptide vasopressin has an important role in this entrainment. Many SCN neurons express vasopressin, and it has been assumed that the role of vasopressin in the SCN reflects the activity of these cells. Here we show that vasopressin is also expressed in many retinal cells that project to the SCN. Light-evoked vasopressin release contributes to the responses of SCN neurons to light, and enhances expression of the immediate early gene c-fos in the SCN, which is involved in photic entrainment of circadian rhythms.

Horizontal cells expressing melanopsin x are novel photoreceptors in the avian inner retina

In the vertebrate retina, three types of photoreceptors-visual photoreceptor cones and rods and the intrinsically photosensitive retinal ganglion cells (ipRGCs)-converged through evolution to detect light and regulate image- and nonimage-forming activities such as photic entrainment of circadian rhythms, pupillary light reflexes, etc. ipRGCs express the nonvisual photopigment melanopsin (OPN4), encoded by two genes: the Xenopus (Opn4x) and mammalian (Opn4m) orthologs. In the chicken retina, both OPN4 proteins are found in ipRGCs, and Opn4x is also present in retinal horizontal cells (HCs), which connect with visual photoreceptors. Here we investigate the intrinsic photosensitivity and functioning of HCs from primary cultures of embryonic retinas at day 15 by using calcium fluorescent fluo4 imaging, pharmacological inhibitory treatments, and Opn4x knockdown. Results show that HCs are avian photoreceptors with a retinal-based OPN4X photopigment conferring intrinsic photosensitivity. Light responses in HCs appear to be driven through an ancient type of phototransduction cascade similar to that in rhabdomeric photoreceptors involving a G-protein q, the activation of phospholipase C, calcium mobilization, and the release of the inhibitory neurotransmitter GABA. Based on their intrinsic photosensitivity, HCs may have a key dual function in the retina of vertebrates, potentially regulating nonvisual tasks together with their sister cells, ipRGCs, and with visual photoreceptors, modulating lateral interactions and retinal processing.

Potent social synchronization can override photic entrainment of circadian rhythms.

Circadian rhythms in behaviour and physiology are important for animal health and survival. Studies with individually isolated animals in the laboratory have consistently emphasized the dominant role of light for the entrainment of circadian rhythms to relevant environmental cycles. Although in nature interactions with conspecifics are functionally significant, social signals are typically not considered important time-givers for the animal circadian clock. Our results challenge this view. By studying honeybees in an ecologically relevant context and using a massive data set, we demonstrate that social entrainment can be potent, may act without direct contact with other individuals and does not rely on gating the exposure to light. We show for the first time that social time cues stably entrain the clock, even in animals experiencing conflicting photic and social environmental cycles. These findings add to the growing appreciation for the importance of studying circadian rhythms in ecologically relevant contexts.

Glutamate-Dependent BMAL1 Regulation in Cultured Bergmann Glia Cells.

Glutamate, the major excitatory amino acid, activates a wide variety of signal transduction cascades. This neurotransmitter is involved in photic entrainment of circadian rhythms, which regulate physiological and behavioral functions. The circadian clock in vertebrates is based on a transcription-translation feedback loop in which Brain and muscle aryl hydrocarbon receptor nuclear translocator (ARNT)-like protein 1 (BMAL1) acts as transcriptional activator of others clock genes. This protein is expressed in nearly all suprachiasmatic nucleus neurons, as well as in the granular layer of the cerebellum. In this context, we decided to investigate the role of glutamate in the molecular mechanisms involved in the processes of transcription/translation of BMAL1 protein. To this end, primary cultures of chick cerebellar Bergmann glial cells were stimulated with glutamatergic ligands and we found that BMAL1 levels increased in a dose- and time dependent manner. Additionally, we studied the phosphorylation of serine residues in BMAL1 under glutamate stimulation and we were able to detect an increase in the phosphorylation of this protein. The increased expression of BMAL1 is most probably the result of a stabilization of the protein after it has been phosphorylated by the cyclic AMP-dependent protein kinase and/or the Ca(2+)/diacylglycerol dependent protein kinase. The present results strongly suggest that glutamate participates in regulating BMAL1 in glial cells and that these cells might prove to be important in the control of circadian rhythms in the cerebellum.

Wiring the retinal circuits activated by light during early development

BackgroundLight information is sorted by neuronal circuits to generate image-forming (IF) (interpretation and tracking of visual objects and patterns) and non-image-forming (NIF) tasks. Among the NIF tasks, photic entrainment of circadian rhythms, the pupillary light reflex, and sleep are all associated with physiological responses, mediated mainly by a small group of melanopsin-expressing retinal ganglion cells (mRGCs). Using Xenopus laevis as a model system, and analyzing the c-fos expression induced by light as a surrogate marker of neural activity, we aimed to establish the developmental time at which the cells participating in both systems come on-line in the retina.ResultsWe found that the peripheral retina contains 80% of the two melanopsin-expressing cell types we identified in Xenopus: melanopsin-expressing horizontal cells (mHCs; opn4m+/opn4x+/Prox1+) and mRGCs (2.7% of the total RGCs; opn4m+/opn4x+/Pax6+/Isl1), in a ratio of 6:1. Only mRGCs induced c-fos expression in response to light. Dopaminergic (tyrosine hydroxylase-positive; TH+) amacrine cells (ACs) may be part of the melanopsin-mediated circuit, as shown by preferential c-fos induction by blue light. In the central retina, two cell types in the inner nuclear layer (INL) showed light-mediated induction of c-fos expression [(On-bipolar cells (Otx2+/Isl1+), and a sub-population of ACs (Pax6−/Isl1−)], as well as two RGC sub-populations (Isl1+/Pax6+ and Isl1+/Pax6−). Melanopsin and opsin expression turned on a day before the point at which c-fos expression could first be activated by light (Stage 37/38), in cells of both the classic vision circuit, and those that participate in the retinal component of the NIF circuit. Key to the classic vision circuit is that the component cells engage from the beginning as functional ‘unit circuits’ of two to three cells in the INL for every RGC, with subsequent growth of the vision circuit occurring by the wiring in of more units.ConclusionsWe identified melanopsin-expressing cells and specific cell types in the INL and the RGC layer which induce c-fos expression in response to light, and we determined the developmental time when they become active. We suggest an initial formulation of retinal circuits corresponding to the classic vision pathway and melanopsin-mediated circuits to which they may contribute.

Phase Misalignment between Suprachiasmatic Neuronal Oscillators Impairs Photic Behavioral Phase Shifts But Not Photic Induction of Gene Expression

The ability of the circadian pacemaker within the suprachiasmatic nucleus (SCN) to respond to light stimulation in a phase-specific manner constitutes the basis for photic entrainment of circadian rhythms. The neural basis for this phase specificity is unclear. We asked whether a lack of synchrony between SCN neurons, as reflected in phase misalignment between dorsomedial (dmSCN) and ventrolateral (vlSCN) neuronal oscillators in the rat, would impact the ability of the pacemaker to respond to phase-resetting light pulses. Light pulses delivered at maximal phase misalignment between the vlSCN and dmSCN oscillators increased expression of Per1 mRNA, regardless of the circadian phase of the dmSCN. However, phase shifts of locomotor activity were only observed when the vlSCN and dmSCN were phase aligned at the time of stimulation. Our results fit a model in which a vlSCN oscillator phase gates its own response to light and in turn relays light information to a dmSCN oscillator. This model predicts that the phase misalignment that results from circadian internal desynchronization could preserve the ability of light to induce gene expression within the master circadian clock but impair its ability to induce behavioral phase shifts.

Absence of Long-Wavelength Photic Potentiation of Murine Intrinsically Photosensitive Retinal Ganglion Cell Firing In Vitro

Melanopsin is an opsin-family photopigment required for photosensitivity of the intrinsically photosensitive retinal ganglion cells (ipRGCs), which subserve photic entrainment of circadian rhythms in mammals. The melanopsin photocycle is presently unknown but is independent of the enzymatic photocycle employed by rhodopsin and cone opsins. Recent experiments have demonstrated that red-light exposure potentiates circadian phase-shifting responses to blue-light stimuli, consistent with the hypothesis that melanopsin functions as a bistable photopigment. To further test this hypothesis, we analyzed ipRGC firing activity in response to 480-nm blue light with or without intervening long-wavelength 620-nm red-light stimulation, using in vitro multielectrode array recording of postnatal day 8 to 10 murine retina. Cell-firing responses to 480-nm light were highly reproducible. No significant potentiating or bleaching effect of intervening subthreshold 620-nm light on ipRGC firing to 480-nm light could be discerned. Further physiologic and biochemical analysis of the ipRGC photoreception is required to reconcile the presence of long-wavelength potentiation at the level of the SCN with its absence in light-induced ipRGC firing.

C.05.03 Rhythm hypotheses of mood and sleep disorders

The neuropeptides pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) are implicated in the photic entrainment of circadian rhythms in the suprachiasmatic nuclei (SCN). We now report that mice carrying a null mutation of the VPAC2 receptor for VIP and PACAP (Vipr2−/−) are incapable of sustaining normal circadian rhythms of rest/activity behavior. These mice also fail to exhibit circadian expression of the core clock genes mPer1, mPer2, and mCry1 and the clock-controlled gene arginine vasopressin (AVP) in the SCN. Moreover, the mutants fail to show acute induction of mPer1 and mPer2 by nocturnal illumination. This study highlights the role of intercellular neuropeptidergic signaling in maintenance of circadian function within the SCN.

Immunohistochemical Evidence of a Melanopsin Cone in Human Retina

Melanopsin, expressed in a subset of intrinsically photosensitive ganglion cells that project to the suprachiasmatic nucleus (SCN), is involved in the photic entrainment of circadian rhythms and other non-image-forming functions (pupil light reflex, masking, acute heart rate response, and alertness). Melanopsin has recently been shown to be a "bireactive" photopigment that functions as a photosensory opsin using 11-cis retinaldehyde as a chromophore and has intrinsic photoisomerase activity. Melanopsin is widely distributed in the retina of vertebrates and, depending on the species, is expressed in ganglion, amacrine, horizontal, and photoreceptor cells. The present study was conducted to determine the distribution of this opsin in the human retina. Human donor eyes were obtained from donors and fixed shortly after death. Immunohistochemistry was used to determine melanopsin expression in the retinas of three donors. The possible coexpression of this photopigment with other opsins was studied by double-labeling immunocytochemistry and confocal analysis. In addition to the expected labeling in ganglion cells of the inner retinal layers, an unexpected finding showed melanopsin-immunopositive label in the outer segments of cones that did not coexpress other known opsins. These melanopsin-expressing cones are extremely sparse (5-25 cones/mm2; 0.1%-0.5% of the entire cone population) and are located in the peripheral retina. The presence of melanopsin in human cones suggests image and non-image-forming roles in visual responses at both the cone input and ganglion cell output stages and their involvement in a broad spectrum of irradiance detection functions in the visual system.

Individual variability and photic entrainment of circadian rhythms in golden spiny mice

Golden spiny mice are diurnally active in most of their natural habitat. Their diurnal activity is ascribed to non-photic cues: competitive exclusion from the nocturnal niche, or thermoregulatory considerations. Here we studied the entrainment of golden spiny mice to light. In the laboratory, golden spiny mice were primarily nocturnal and displayed an unusual variety of rhythm patterns, with activity bursts occurring during both activity and rest periods. Spontaneous shifts of activity rhythms between light phases were sometimes recorded. In all cases but one, body temperature shifted in parallel with activity. Under DD conditions, the free running period (tau) of all individuals but one was shorter than 24 h, and in all individuals but the same one it was shorter than tau under LL conditions. In response to a 6 h phase delay, all individuals entrained to the new LD cycle in a relatively uniform way. During phase advance four out of the twelve individuals further delayed their activity and body temperature rhythms, and eight individuals advanced their activity rhythm, but the re-entrainment took them over twice as long as to re-entrain to the phase delay. We suggest that the golden spiny mouse is a nocturnal rodent whose circadian system developed the flexibility to be nocturnal or diurnal according to environmental conditions, or a nocturnal rodent in the process of turning diurnal, and that it has low sensitivity to the immediate masking effect of light on activity.

Light response of the neuronal firing activity in the suprachiasmatic nucleus of mice

To investigate the neural mechanisms underlying the mammalian photic entrainment of circadian rhythms, the response of neuronal extracellular firing activity to retinal light stimulation was investigated in the suprachiasmatic nucleus (SCN) of anesthetized mice during nighttime and daytime. In nighttime, most recorded SCN cells (83%) increased their firing frequency in response to retinal illumination. Some SCN cells (11%) responded by decreasing their firing rate. In daytime, the retinal illumination increased the firing rate in only 26% of the SCN cells, and no response was observed in the remaining cells. The light intensity threshold for the activation of SCN cells at zeitgeber time (ZT) 16 was approximately 3 × 10 11 photons cm −2 s −1 and the maximum response was observed at approximately 1 × 10 14 photons cm −2 s −1. Therefore, photic response in the firing of mouse SCN cells may be phase-dependent and have a higher threshold, which corresponds to properties of the photic entrainment in locomotor activity of mice.

Fos expression in the suprachiasmatic nucleus during photic entrainment of circadian rhythms in retinally damaged rats.

The protein product of the immediate-early gene c-fos is expressed rhythmically in the shell region of the suprachiasmatic nucleus (SCN), the mammalian circadian clock. Recently, we found that exposure to an entraining light pulse caused a suppression of Fos expression in the SCN shell in rats. To study the hypothesis that suppression of Fos in the shell is a correlate of photic entrainment, we used rats that were treated with the retinal neurotoxin monosodium glutamate (MSG) during the neonatal period. In spite of retinal degeneration, MSG-treated rats entrained normally and displayed light-induced suppression of Fos within the SCN shell. These results support the view that light-induced suppression of Fos within the SCN shell is a cellular correlate of photic entrainment.

Light response of the neuronal firing activity in the suprachiasmatic nucleus of mice

Abstract To investigate the neural mechanisms underlying the mammalian photic entrainment of circadian rhythms, the response of neuronal extracellular firing activity to retinal light stimulation was investigated in the suprachiasmatic nucleus (SCN) of anesthetized mice during nighttime and daytime. In nighttime, most recorded SCN cells (83%) increased their firing frequency in response to retinal illumination. Some SCN cells (11%) responded by decreasing their firing rate. In daytime, the retinal illumination increased the firing rate in only 26% of the SCN cells, and no response was observed in the remaining cells. The light intensity threshold for the activation of SCN cells at zeitgeber time (ZT) 16 was approximately 3 × 10 11 photons cm −2 s −1 and the maximum response was observed at approximately 1 × 10 14 photons cm −2 s −1 . Therefore, photic response in the firing of mouse SCN cells may be phase-dependent and have a higher threshold, which corresponds to properties of the photic entrainment in locomotor activity of mice.

Illuminating the circadian clock in monarch butterfly migration.

Migratory monarch butterflies use a time-compensated Sun compass to navigate to their overwintering grounds in Mexico. Here, we report that constant light, which disrupts circadian clock function at both the behavioral and molecular levels in monarchs, also disrupts the time-compensated component of flight navigation. We further show that ultraviolet light is important for flight navigation but is not required for photic entrainment of circadian rhythms. Tracing these distinct light-input pathways into the brain should aid our understanding of the clock-compass mechanisms necessary for successful migration.

Effect of 192 IgG-Saporin on Circadian Activity Rhythms, Expression of P75 Neurotrophin Receptors, Calbindin-D28K, and Light-Induced Fos in the Suprachiasmatic Nucleus in Rats

Photic entrainment of circadian rhythms in mammals is mediated through a direct retinal projection to the core region of the suprachiasmatic nucleus (SCN), the circadian clock. A proportion of this projection contains the low-affinity p75 neurotrophic receptor (p75NTR). Neonatal monosodium glutamate (MSG) treatment, which dramatically reduces p75NTR immunoreactivity in the SCN has no impact on photic entrainment. In order to clarify the contribution of p75NTR fibers in photic entrainment, targeted lesions of the p75NTR-immunoreactive SCN plexus were performed using intracerebroventricular (ICV) or intrahypothalamic injections of the immunotoxin 192 IgG-saporin (SAP) in rats. SAP treatment effectively abolished p75NTR immunoreactivity within the SCN core. ICV SAP treatment produced three different behavioral activity patterns: Animals became arrhythmic, displayed a shorter free-running period, or remained rhythmic following the lesion. Arrhythmic animals had large hypothalamic lesion which encompassed the entire SCN. In rhythmic rats, ICV-SAP significantly reduced immunostaining for calbindin-D28k (CaBP) in the SCN, and rats with shortened free-running periods had the lowest number of CaBP immunoreactive cells. ICV SAP also attenuated light-induced Fos expression in the SCN core. Despite lack of p75NTR and reduced CaBP and Fos expression in the SCN, SAP-treated rhythmic rats displayed normal photic entrainment. Intrahypothalamic SAP treatment reduced CaBP expression in the SCN but had no effect on light-induced Fos expression, free-running rhythms, or photic entrainment. The data show that p75NTR-immunoreactive elements in the SCN are not required for photic entrainment.

The VPAC 2 Receptor Is Essential for Circadian Function in the Mouse Suprachiasmatic Nuclei

The neuropeptides pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) are implicated in the photic entrainment of circadian rhythms in the suprachiasmatic nuclei (SCN). We now report that mice carrying a null mutation of the VPAC 2 receptor for VIP and PACAP ( Vipr2 −/− ) are incapable of sustaining normal circadian rhythms of rest/activity behavior. These mice also fail to exhibit circadian expression of the core clock genes mPer1, mPer2, and mCry1 and the clock-controlled gene arginine vasopressin (AVP) in the SCN. Moreover, the mutants fail to show acute induction of mPer1 and mPer2 by nocturnal illumination. This study highlights the role of intercellular neuropeptidergic signaling in maintenance of circadian function within the SCN.

Light suppresses Fos expression in the shell region of the suprachiasmatic nucleus at dusk and dawn: implications for photic entrainment of circadian rhythms

The transcription factor Fos is implicated in neuronal signaling in the suprachiasmatic nucleus, the mammalian circadian clock ( Ikonomov and Stoynev, 1994; Klein et al., 1991; Kornhauser et al., 1996). Fos is expressed in two different regions within the suprachiasmatic nucleus. In the ventrolateral, retinorecipient, core region ( Leak et al., 1999; Moga and Moore, 1997), Fos is induced by light and expression is closely linked, both temporally and functionally, to clock resetting and entrainment of circadian rhythms ( Aronin et al., 1990; Beaulé and Amir, 1999; Hastings et al., 1995; Kornhauser et al., 1996; Kornhauser et al., 1990; Rea, 1989, 1998; Rusak et al., 1990; Wollnik et al., 1995). In the dorsomedial shell region ( Leak et al., 1999), Fos expression is rhythmic ( Guido et al., 1999a,b; Rusak et al., 1992; Sumova and Illnerova, 1998; Sumova et al., 1998). Expression is high during the subjective day when photic sensitivity of the core is minimal, and low in the subjective night, when photic sensitivity of the core is maximal. Although it has been shown that the pattern of Fos expression in the shell tracks the photoperiod ( Sumova et al., 2000), nothing is known about whether light influences the expression of Fos in the shell region or about the role of Fos expression in the shell in clock resetting and entrainment. In the present study we found that, in rats maintained in constant darkness, brief exposure to light in the early subjective day or night induced Fos in the core, as expected, and acutely suppressed the levels of Fos immunoreactivity in the shell region. Similar changes in Fos expression in the core and shell regions were seen after exposure to a brief entraining light. Light exposure in the mid-subjective day or night differentially affected Fos expression in the core, as previously shown, but had no effect on Fos expression in the shell region. The finding that Fos expression in the shell region of the suprachiasmatic nucleus is suppressed by light at dawn and dusk suggests a critical role for the shell in photic entrainment of circadian rhythms in nocturnal rodents.

Photic regulation of circadian rhythms and the expression of p75 neurotrophin receptor immunoreactivity in the suprachiasmatic nucleus in rats

Neurotrophic factors have been implicated in the mechanism underlying photic regulation of circadian rhythms in mammals. In rats, the most abundant neurotrophin receptor found in the suprachiasmatic nucleus (SCN), the circadian clock, is the low affinity p75 neurotrophin receptor (p75NTR). This receptor is expressed by retinal afferents of the SCN, but nothing is known about its role in photic regulation of circadian rhythms. We show here that neonatal treatment with the retinal neurotoxin, monosodium glutamate (MSG), which has no effect on photic entrainment of circadian rhythms, nearly completely abolished p75NTR immunoreactivity in the SCN in rats. These findings suggest that p75NTR from retinal sources do not play an essential role in the mechanism mediating photic entrainment of circadian rhythms in rats.

On the Use of the Time Axis for Ecological Separation: Diel Rhythms as an Evolutionary Constraint

On the Use of the Time Axis for Ecological Separation: Diel Rhythms as an Evolutionary Constraint

Is light-regulated AP-1 binding in the rat suprachiasmatic nucleus gated by the circadian clock?

In mammals, photic entrainment of circadian rhythms likely involves light- and clock-dependent expression of immediate early genes, including fos-like and jun-like genes, in the rat suprachiasmatic nucleus. Using an electrophoretic mobility shift assay, we evaluated whether the photic regulation of DNA-binding activity and composition of activating protein-1 (AP-1) complexes in the suprachiasmatic nucleus is also dependent on circadian phase. Phase-dependent light inducibility in the expression of fra-2 and c-fos genes and in immunoreactive Fra-2 and c-Fos protein expression was also evaluated, by in situ hybridization and immunocytochemistry. Light’s effects on AP-1 DNA-binding differed both qualitatively and quantitatively according to the circadian phase at which light was applied. This phase dependence accounted for by both compartmentalization of proteins involved in constitutive AP-1 complexes within the nucleus or cytoplasm and control of the extent to which the expression of specific complexes was induced. It was then shown that the mechanisms by which the circadian clock gates the photic induction of AP-1 components differed according to the nature of the protein.