Photic Phase Shifts Research Articles

Daily Rhythms of Liver-Function Indicators in Rabbits

Serum concentrations of urea and cholesterol were used as indicators of liver function in rabbits in an investigation of the responsiveness of the daily rhythm of liver function to phase shifts in the environmental light-dark cycle. Locomotor activity was simultaneously monitored by actigraphy. Serum urea and cholesterol concentrations exhibited robust daily rhythmicity with opposite phases. Both indicators of liver function phase shifted in response to phase shifts of the light-dark cycle. The phase shifts in liver function appeared to be parallel to phase shifts in locomotor activity, but the data were not sufficiently strong to confirm that the liver relies on the secondary effect of photic phase shift on ingestive behavior in order to be phase-shifted by light as previously indicated by gene expression studies in laboratory rodents.

Inhibiting cortisol response accelerates recovery from a photic phase shift.

Jetlag results when a temporary loss of circadian entrainment alters phase relationships among internal rhythms and between an organism and the outside world. After a large shift in the light-dark (LD) cycle, rapid recovery of entrainment minimizes the negative effects of internal circadian disorganization. There is evidence in the existing literature for an activation of the hypothalamic-pituitary-adrenal (HPA) axis after a photic phase shift, and it is possible that the degree of HPA-axis response is a determining factor of reentrainment time. This study utilized a diurnal rodent, Octodon degus, to test the prediction that the alteration of cortisol levels would affect the reentrainment rate of circadian locomotor rhythms. In experiment 1, we examined the effects of decreased cortisol (using metyrapone, an 11beta-hydroxylase inhibitor) on the rate of running-wheel rhythm recovery after a 6-h photic phase advance. Metyrapone treatment significantly shortened the length of time it took animals to entrain to the new LD cycle (11.5% acceleration). In experiment 2, we examined the effects of increased cortisol on the rate of reentrainment after a 6-h photic phase advance. Increasing plasma cortisol levels increased the number of days (8%) animals took to reentrain running-wheel activity rhythms, but this effect did not reach significance. A third experiment replicated the results of experiment 1 and also demonstrated that suppression of HPA activity via dexamethasone injection is capable of accelerating reentrainment rates by approximately 33%. These studies provide support for an interaction between the stress axis and circadian rhythms in determining the rate of recovery from a phase shift of the LD cycle.

Neuropeptide Y attenuates NMDA-induced phase shifts in the SCN of NPY Y1 receptor knockout mice in vitro

Neuropeptide Y (NPY) blocks the effect of light on the mammalian circadian clock during the subjective night. The present study explores the role of the NPY Y1 receptor in this interaction. The effect of NPY when co-applied with NMDA, a glutamate agonist that can mimic the effect of light, was examined in NPY Y1−/− mice (background strain 129SVXBalb/c) using electrophysiology. Cells in the suprachiasmatic nucleus (SCN), the master circadian pacemaker, show a circadian rhythm in spontaneous firing rate that can be recorded in vitro. The results demonstrated that NPY attenuated the phase shifts to NMDA in both the Y1−/− mice and control mice, indicating that the Y1 receptor does not mediate the NPY blockade of photic-like phase shifts. The peak in frequency in the untreated control brain slices from Y1−/− mice was advanced by approximately 1 h as compared to the Y1+/+ mice. The Y1 receptor may contribute to a functional model of circadian rhythms, but apparently is not essential for the effects of NPY on photic phase shifts.

Blockade of the NPY Y5 receptor potentiates circadian responses to light: complementary in vivo and in vitro studies.

Neuropeptide Y (NPY) is delivered to the suprachiasmatic nuclei (SCN) circadian pacemaker via an input from the thalamic intergeniculate leaflet. NPY can inhibit light-induced responses of the circadian system of Syrian hamsters. Here we studied whether an antagonist to NPY receptors can be used to potentiate photic phase shifts late in the subjective night. First we determined by in situ hybridization that both NPY Y1 and Y5 receptor mRNA are expressed in the SCN of Syrian hamsters. Second, similar to our previous findings at Zeitgeber time 14 (ZT 14, where ZT 12 was the time of lights off), we found that NPY applied at ZT 18.5 onto the SCN region of brain slices maintained in vitro could block NMDA-induced phase advances of the spontaneous firing rate rhythm, and this blocking effect was probably mediated by the Y5 receptor, since co-application of Y5 receptor antagonists completely reversed the effect of NPY, while application of a Y1 receptor antagonist had no effect under the same conditions. Third, we found that co-treatment with a Y5 receptor antagonist in vivo (s.c., 10 mg/kg) not only reversed the effect of NPY applied to the SCN in vivo through a cannula but also significantly potentiated the light-induced phase advance in the absence of NPY. This is the first report of a NPY receptor antagonist having such an effect, and indicates that NPY Y5 receptor antagonists could be clinically useful for potentiating circadian system responses to light.

Neuropeptide y, gaba and circadian phase shifts to photic stimuli

Circadian rhythms can be phase shifted by photic and non-photic stimuli. The circadian clock, anatomically defined as the suprachiasmatic nucleus (SCN), can be phase delayed by light during the early subjective night and phase advanced during the late subjective night. Non-photic stimuli reset the clock when presented during the subjective day. A possible pathway for the non-photic resetting of the clock is thought to originate from the intergeniculate leaflet, which conveys information to the SCN through the geniculohypothalamic tract and utilizes among others neuropeptide Y (NPY) and GABA as neurotransmitters. Photic and non-photic stimuli have been shown to interact during the early and late subjective night. Microinjections of NPY or muscimol, a GABAA receptor agonist, into the region of the SCN can attenuate light-induced phase shifts during the early and late subjective night. The precise mechanism for these interactions is unknown.In the current study we investigate the involvement of a GABAergic mechanism in the interaction between NPY and light during the early and late subjective night. Microinjections of NPY significantly attenuated light-induced phase delays and inhibited phase advances (P<0.05). The administration of bicuculline during light exposure, before NPY microinjection did not alter the ability of NPY to attenuate light-induced phase delays and block photic phase advances.These results indicate that NPY attenuates photic phase shifts via a mechanism independent of GABAA receptor activation. Furthermore it is evident that NPY influences circadian clock function via differing cellular pathways over the course of a circadian cycle.

Attenuation of circadian light induced phase advances and delays by neuropeptide y and a neuropeptide y y1/y5 receptor agonist

Circadian rhythms can be synchronised to photic and non-photic stimuli. The circadian clock, anatomically defined as the suprachiasmatic nucleus in mammals, can be phase shifted by light during the night. Non-photic stimuli reset the circadian rhythm during the day. Photic and non-photic stimuli have been shown to interact during the day and night. Precise mechanisms for these complex interactions are unknown. A possible pathway for non-photic resetting of the clock is thought to generate from the intergeniculate leaflet, which conveys information to the suprachiasmatic nucleus (SCN) through the geniculohypothalamic tract and utilises neuropeptide Y (NPY) as its primary neurotransmitter. Interactions between light and NPY were investigated during the early (2 h after activity onset) and late (6 h after activity onset) night in male Syrian hamsters. NPY microinjections into the region of the SCN significantly attenuated light-induced phase delay, during the early subjective night. Phase advances to light were completely inhibited by the administration of NPY during the late night. The precise mechanism by which NPY attenuates or blocks photic phase shifts is unclear, but the NPY Y5 receptor has been implicated in the mediation of this inhibitory effect. The NPY Y1/Y5 receptor agonist, [Leu 31,Pro 34]NPY, was administered via cannula microinjections following light exposure during the early and late night. [Leu 31,Pro 34]NPY significantly attenuated phase delays to light during the early night and blocked phase advances during the late night, in a manner similar to NPY. These results show the ability of NPY to attenuate phase shifts to light during the early night and block light-induced phase advances during the late night. Furthermore, this is the first in vivo study implicating the involvement of the NPY Y1/Y5 receptors in the complex interaction of photic and non-photic stimuli during the night. The alteration of photic phase shifts by NPY may influence photic entrainment within the circadian system.

Running wheel size influences circadian rhythm period and its phase shift in mice.

Running wheels are widely used in studies on biological rhythms. In mice wheel diameters have ranged from 11 cm to 23 cm. We provided mice with running wheels of two different sizes: 15 cm diameter and 11 cm diameter. The amount of running in the 12-h light:12-h dark condition and the endogenous period of wheel running in constant darkness was determined over 40 days. On the 1st day in constant darkness all animals were exposed to a 15-min light pulse at circadian time 13. The animals in the small wheel ran significantly less both in 12 h light: 12 h dark and constant darkness, and showed a longer endogenous period in constant darkness compared to animals in the large wheel. Moreover, after the light pulse at circadian time 13, mice in the small wheel showed a significantly smaller phase delay in running wheel activity than mice in the larger wheels. The data suggest that the magnitude of a photic phase shift depends on the amount and timing of activity the animals display in relation to this stimulus. It can be concluded that technical features of the running wheel can influence the circadian period of wheel running.

Serotonergic Modulation of Photic Entrainment in the Syrian Hamster

In this review, we describe six lines of evidence that reveal a modulatory role for serotonin (5-HT) in the regulation of the response of suprachiasmatic nucleus (SCN) neurons to retinal illumination in the Syrian hamster. Electrical stimulation of the median raphe nucleus, sufficient to elicit the release of 5-HT in the SCN, inhibits light-induced phase shifts of the hamster circadian activity rhythm. Two 5-HT receptors capable of mediating the effects of 5-HT on photic responses, the 5-HT7 receptor and the 5-HT1B receptor, are present in the hamster SCN. Light-induced phase shifts are attenuated by systemic and local administration of two 5-HT receptor agonists, 8-OH-DPAT, and TFMPP, and these agents attenuate photic phase shifts by acting on pharmacologically distinct receptors. Furthermore, both compounds also attenuate light-induced Fos expression and photic suppression of pineal melatonin content, indicating that serotonergic modulation of photic signal transduction in the SCN is not limited to the regulation of circadian phase. Finally, both 8-OH-DPAT and TFMPP inhibit RHT neurotransmission in the hypothalamic slice preparation. Further, TFMPP fails to attenuate responses to exogenous glutamate on retinorecipient SCN neurons, consistent with a presynaptic site of action for the drug. Based on these data, we propose that 5-HT modulates RHT neurotransmission in the SCN through at least two distinct mechanisms: (1) via activation of 5-HT7 receptors probably located on retinorecipient neurons; and (2) via activation of presynaptic 5-HT1B receptors leading to reduced release of glutamate from RHT terminals in the SCN.

Is there a geniculohypothalamic tract in primates? A comparative immunohistochemical study in the circadian system of strepsirhine and haplorhine species

In rodents, the circadian rhythm generated by the hypothalamic suprachiasmatic nucleus (SCN) is modulated by two types of phenomena: photic phase-shifts, mediated by the retinohypothalamic pathway and non-photic phase-shifts mediated by the projection of the intergeniculate leaflet (IGL) to the SCN which contains the neuropeptide Y (NPY). In primates, the retinohypothalamic pathway has been well-demonstrated but very little is known about the geniculohypothalamic tract. This prompted us to study NPY immunoreactivity in both the SCN and the IGL in species representative of the three main primate lineages: prosimians ( Microcebus), New World monkeys ( Callithrix) and Old World monkeys ( Macacca). In species studied, we found a region in the pregeniculate nucleus containing both NPY immunopositive cells and substance P immunopositive fibres that we identified as the IGL. During evolution, this structure has moved from a ventral to a dorsomedial position relative to the adjacent dorsal lateral geniculate nucleus. By contrast, NPY-IP fibres in the SCN are dense in prosimians, but are sparse or absent in other primate species. We suggest that either the geniculohypothalamic projection is absent in higher primates as is the case in humans, or is absent in diurnal mammals, or contains a different peptide, or that NPY immunoreactivity varies according to other parameters.

Serotonin-containing fibers in the suprachiasmatic hypothalamus attenuate light-induced phase delays in mice

Photic and non-photic stimuli phase shift and entrain circadian rhythms through distinct but interacting mechanisms which impinge on the suprachiasmatic nucleus (SCN), the circadian pacemaker. Our understanding of this mechanism is incomplete. Serotonin (5-HT) injected locally at the SCN reduces light-induced glutamate release and decreases the expression of c- fos, a marker of photic transduction. Furthermore, in SCN slices, 5-HT application reduces field potentials after optic nerve stimulation. We therefore predicted that 5-HT-terminal destruction restricted to the SCN would augment phase shifts of circadian rhythms induced by light exposure. To investigate this possibility, we compared photic phase delays and Fos-like immunoreactivity in mice which had previously received bilateral infusions directed at the SCN containing either the selective 5-HT neurotoxin 5,7-dihydroxytryptamine (DHT, n=16) or vehicle (VEH, n=12). Phase delays after a light pulse given during the mid-subjective night (30 lux, 30 min starting at circadian time (CT) 12–20) in DHT-mice were 50% greater than in VEH-mice ( P=0.017). DHT mice ( n=5) had 76% larger Fos responses to a mid-subjective night light pulse than VEH-mice ( n=5) ( P=0.029). We conclude that 5-HT at or near the SCN in mice reduces photic phase shifts and modulates the magnitude of the photic phase response in the mouse.

Neuropeptide Y and glutamate block each other's phase shifts in the suprachiasmatic nucleus in vitro

The suprachiasmatic nuclei contain a circadian clock whose activity can be recorded in vitro for several days. Photic information is conveyed to the nuclei primarily via a direct projection from the retina, the retinohypothalamic tract, utilizing an excitatory amino acid neurotransmitter. Photic phase shifts may be mimicked by application of glutamate in vitro. A second, indirect pathway to the suprachiasmatic nuclei via the geniculohypothalamic tract utilizes neuropeptide Y as a transmitter. Phase shifts to neuropeptide Y in vitro are similar to those seen to non-photic stimuli in vivo. We have used the hypothalamic slice preparation to examine the interactions of photic and non-photic stimuli in the suprachiasmatic nuclei. Coronal hypothalamic slices containing the suprachiasmatic nuclei were prepared from Syrian hamsters and 3 min recordings of the firing rate of individual cells were performed throughout a 12 h period. Control slices receiving either no application or application of artificial cerebrospinal fluid to the suprachiasmatic nucleus showed a consistent daily peak in their rhythms. Glutamate produces phase shifts of the circadian clock in the hamster hypothalamic slice preparation during the subjective night but not during the subjective day. These phase shifts were similar in timing and direction to the photic phase response curve in vivo confirming previous work with the rat slice preparation. Neuropeptide Y produces phase shifts of the circadian clock during the subjective day but not during the subjective night. The phase shifts are similar in timing and direction to the non-photic phase response curve in vivo, confirming previous in vitro work. We then examined the interaction of these neurochemicals with each other at various times during the circadian cycle. We found that both advances and delays to glutamate in the slice are blocked by application of neuropeptide Y. We also found that phase shifts to neuropeptide Y in the slice are blocked by application of glutamate. These results indicate that photic and non-photic associated neurochemicals can block each others phase shifting effects within the suprachiasmatic nucleus in vitro. These experiments demonstrate the ability of photic and non-photic associated neurochemicals to interact at the level of the suprachiasmatic nucleus. It is clear that neuropeptide Y antagonizes the effect of glutamate during the subjective night, and that glutamate antagonizes the effect of neuropeptide Y during the subjective day. Great care must be taken when devising treatments where photic and non-photic signals may interact.

The Ventral Lateral Geniculate Nucleus and the Intergeniculate Leaflet: Interrelated Structures in the Visual and Circadian Systems

HARRINGTON, M.E. The ventral lateral geniculate nucleus and the intergeniculate leaflet: interrelated structures in the visual and circadian systems. NEUROSCI BIOBEHAV REV 21(5) 705–727 1997.—The ventral lateral geniculate nucleus (vLGN) and the intergeniculate leaflet (IGL) are retinorecipient subcortical nuclei. This paper attempts a comprehensive summary of research on these thalamic areas, drawing on anatomical, electrophysiological, and behavioral studies. From the current perspective, the vLGN and IGL appear closely linked, in that they share many neurochemicals, projections, and physiological properties. Neurochemicals commonly reported in the vLGN and IGL are neuropeptide Y, GABA, enkephalin, and nitric oxide synthase (localized in cells) and serotonin, acetylcholine, histamine, dopamine and noradrenalin (localized in fibers). Afferent and efferent connections are also similar, with both areas commonly receiving input from the retina, locus coreuleus, and raphe, having reciprocal connections with superior colliculus, pretectum and hypothalamus, and also showing connections to zona incerta, accessory optic system, pons, the contralateral vLGN/IGL, and other thalamic nuclei. Physiological studies indicate species differences, with spectral-sensitive responses common in some species, and varying populations of motion-sensitive units or units linked to optokinetic stimulation. A high percentage of IGL neurons show light intensity-coding responses. Behavioral studies suggest that the vLGN and IGL play a major role in mediating non-photic phase shifts of circadian rhythms, largely via neuropeptide Y, but may also play a role in photic phase shifts and in photoperiodic responses. The vLGN and IGL may participate in two major functional systems, those controlling visuomotor responses and those controlling circadian rhythms. Future research should be directed toward further integration of these diverse findings. © 1997 Elsevier Science Ltd.

Photoperiod-dependent correlation between light-induced SCN c-fos expression and resetting of circadian phase.

In rodents, brief light pulses that shift the phase of the circadian locomotor rhythm also increase c-fos gene expression in the suprachiasmatic nucleus (SCN), site of an endogenous clock that regulates such rhythmicity. Because the magnitude of photic phase shifts varies when light pulses are applied at different time points over the course of the subjective night, we examined the degree of SCN c-fos gene expression after single 30-min light pulses were delivered at time points that spanned the early and late subjective night in rats maintained in either short (8:16-h light-dark cycle) or long (16:8-h light-dark cycle) photoperiods. The light-induced level of c-fos mRNA and the number of cells expressing immunoreactive c-Fos protein were measured in the SCN by in situ hybridization and immunohistochemistry, respectively, and compared with the magnitude of the corresponding phase shifts of the circadian rhythm of pineal N-acetyltransferase (NAT) activity. We found a robust correlation between c-fos photoinduction and NAT phase shifts, but this relationship was dependent on photoperiod. The degree of c-fos gene expression was strongly correlated with the magnitude of NAT phase advances and delays under the short photoperiod and with phase advances under the long photoperiod, but not with phase delays under the long photoperiod. The data suggest that c-fos gene expression in the SCN may be involved in the photic resetting of the pineal NAT rhythm. Under the long photoperiod, however, the magnitude of phase delays may be limited by the functional state of the circadian pacemaking system.

Circadian Phase Shifting Associated with Routine Cage Maintenance: A Retrospective Analysis

Stimuli that evoke behavioral activation can phase-shift free-running circadian activity rhythms in Syrian hamsters. Activation-induced phase shifting is characterized by a phase-response curve (PRC) that is dissimilar to the PRC for photic phase shifting, and recent studies indicate that complex interactions may occur between photic and non-photic phase shifting. Since animals in the laboratory may be exposed to both photic and behaviorally activating stimulation during routine cage maintenance procedures, we performed a retrospective analysis of possible phase shifts associated with cage cleaning in individually housed hamsters maintained in either constant darkness (DD) or dim red light (RR) during the course of an ongoing study of drug-induced phase shifting. All cage cleanings were conducted under RR and were separated from drug treatments by at least one week. The results indicated that both photic and non-photic phase shifts could be induced by routine cage maintenance procedures, depending on the ...

Circadian rhythm photic phase shifts are not altered by histamine receptor antagonists.

Histamine may play a role in synchronizing endogenous circadian rhythms with exogenous photic cues. Direct application of histamine to the suprachiasmatic nucleus, the site of the mammalian circadian pacemaker, phase shifts the circadian rhythm in neural activity [7]. Intraventricular injections of histamine also phase shift circadian rhythms [14]. The magnitude and direction of the phase shifting effects of histamine depend on circadian phase in a manner similar to light [7,14]. Depletion of brain histamine levels by inhibition of histamine synthesis reduces phase shifts to light [10].

Histamine synthesis inhibition reduces light-induced phase shifts of circadian rhythms

Circadian rhythms are generated by the hypothalamic suprachiasmatic nuclei, a site of dense histaminergic innervation. Histamine can phase shift circadian rhythms in a manner similar to light. In this experiment, we administered α-fluoromethylhistidine (FMH), an inhibitor of histamine synthesis, prior to a light pulse in hamsters housed under constant darkness. Photic phase shifts in wheel-running rhythms were significantly attenuated by FMH pretreatment. These results suggest that histamine may modulate photic input to the circadian clock.

CGMP-dependent protein kinase inhibitor blocks light-induced phase advances of circadian rhythms in vivo

The suprachiasmatic nucleus (SCN) contains the primary mammalian circadian clock. Light synchronizes these circadian rhythms through a mechanism involving the release of excitatory amino acids (EAA) and synthesis of nitric oxide (NO) in the SCN. In the current study, we investigated whether cGMP-mediated activation of cGMP-dependent protein kinase (PKG) is associated with light-induced phase shifts of the circadian oscillator. Local administration of the specific PKG inhibitor, KT-5823, significantly attenuated light-induced advances in the phase of activity rhythms when administered during late subjective night (CT 19). Similar treatment at CT 14 had no significant effect on light-induced phase delays. These results are the first to implicate PKG in the biochemical pathway(s) responsible for photic phase advances, and suggest a divergence in biochemical pathways involved in photic phase shifts.

Serotonergic potentiation of photic phase shifts of the circadian activity rhythm.

Recent evidence suggests that serotonin may function to regulate the sensitivity of the circadian clock to the resetting effect of environmental light. Here we report that systemic administration of NAN-190, a drug that acts at both postsynaptic and somatodendritic serotonin receptors, potentiates light-induced phase shifts by as much as 250%. The effects of the drug are dose-related and are significant at light intensities between 0.2 and 200 lux. It is proposed that drugs with pharmacological properties similar to NAN-190 may prove useful as chronobiologics to adjust the sensitivity of the circadian system to natural 'zeitgebers'.

Enhanced photic phase shifting after treatment with antiserum to neuropeptide Y

Previous work has shown that antiserum to neuropeptide Y blocks phase shifts to certain non-photic stimuli at circadian time 4. To examine the role of NPY in photic phase shifts, antiserum to neuropeptide Y was administered just prior to a light pulse at circadian time 18. Hamsters were implanted with guide cannulae aimed at the suprachiasmatic nucleus, and housed in constant darkness. Animals were injected at circadian time 18 under a dim red safe light with 200 nl of either antiserum to neuropeptide Y or normal serum. They were then either exposed to light (approximately 100 lux) for 15 min or returned to constant darkness. Hamsters shifted more to the light pulse when pretreated with antiserum to neuropeptide Y (mean normal serum + light = 1.18 h; mean antiserum to neuropeptide Y + light = 1.92 h; P < 0.01 on a two-tailed paired t-test). Therefore, antiserum to neuropeptide Y enhanced photic advances at circadian time 18.

Double-Pulse Experiments with Nonphotic and Photic Phase-Shifting Stimuli

Three-hour pulses of novelty-induced wheel running in the early to middle subjective day of golden hamsters produced phase advances of 2-3 hr. This phase shifting could be almost totally abolished by a light pulse following within 3 hr of the exercise pulse. When light pulses occurred about 8 hr after the exercise pulses, the phase-advancing effects of the latter were enhanced. Consideration of the amplitude of the phase response curve (PRC) for light pulses alone, in the test paradigms used here, showed that nonphotic and photic phase shifts did not combine additively. Antagonistic and synergistic interactions between photic and nonphotic shifts may have to be taken into account if it transpires that exercise in people can be used to assist adjustment to new schedules after crossing time zones, or in shiftwork.