Induced Phase Delays Research Articles

Vacuum Birefringence Measurement via All-Optical Interferometric Schemes

All-optical vacuum birefringence experiments will get increasingly closer to feasibility as multi-petawatt laser facilities become operational around the World. Thus, the availability of focused laser intensities in the order of IL ∼ 1022 −1024 W/cm2 are to be expected in a focused spot size ∼ 3 − 5 μm. With these values, vacuum refraction indices in the order of Δn ∼ 10−11 − 10−9 are possible with an induced phase delay on a counterpropagating optical probe beam in the order of Δφ ∼ 10−9 −10−7 radians. We discuss two all-optical interferometric schemes and detail the Mach-Zender interferometric proposal. We consider this interferometric scheme fed by both classical and non-classical input light and with two detection schemes. We outline scenarios that are likely to lead to a feasible experimental implementation.

Hormonal fine-tuning of clock in decidual region of mouse placenta by dopamine, melatonin, insulin, leptin and ghrelin

IntroductionThe maternal part of the rodent placenta harbors a circadian clock which robustly responds to glucocorticoids, however, its sensitivity to other hormones has not been elucidated. In this study, we tested five selected hormones (dopamine, melatonin, insulin, leptin and ghrelin) for their effectiveness to affect the clock in decidual region of mouse placenta in vitro. MethodsWe administered the hormones or corresponding vehicles at various time points over 24 h to organotypic placental explants of mPer2Luc mice containing the decidua basalis (DB) region and monitored their effects on amplitude, period, median expression level (mesor) and phase of PER2-driven bioluminescence rhythms. ResultsDopamine significantly increased the amplitude, robustly dampened the mesor, and during a narrow time interval (corresponding to daytime) induced phase delays of the rhythms. In contrast, melatonin had no effect on amplitude, but induced phase advances of the rhythms at the opposite time window than dopamine (corresponding to nighttime). Leptin and ghrelin, but not insulin, slightly increased amplitudes and moderately modulated phase delays of the clock, suggesting that the DB clock, in contrast to other peripheral clocks, is rather resilient to abrupt changes in levels of feeding- and metabolism-related hormones. DiscussionThe results demonstrate for the first time that dopamine and melatonin exhibit delicate yet specific effects on parameters of the DB clock and may thus potentially contribute to fine-tuning of its phase under in vivo conditions. It also implies that dysregulation of their levels, which accompany various pathologies, may account for malfunction of the clock in DB.

TRESK is a key regulator of nocturnal suprachiasmatic nucleus dynamics and light adaptive responses

The suprachiasmatic nucleus (SCN) is a complex structure dependent upon multiple mechanisms to ensure rhythmic electrical activity that varies between day and night, to determine circadian adaptation and behaviours. SCN neurons are exposed to glutamate from multiple sources including from the retino-hypothalamic tract and from astrocytes. However, the mechanism preventing inappropriate post-synaptic glutamatergic effects is unexplored and unknown. Unexpectedly we discovered that TRESK, a calcium regulated two-pore potassium channel, plays a crucial role in this system. We propose that glutamate activates TRESK through NMDA and AMPA mediated calcium influx and calcineurin activation to then oppose further membrane depolarisation and rising intracellular calcium. Hence, in the absence of TRESK, glutamatergic activity is unregulated leading to membrane depolarisation, increased nocturnal SCN firing, inverted basal calcium levels and impaired sensitivity in light induced phase delays. Our data reveals TRESK plays an essential part in SCN regulatory mechanisms and light induced adaptive behaviours.

Quantitative Phase Imaging Camera With a Weak Diffuser

We introduce the quantitative phase imaging camera with a weak diffuser (QPICWD) as a practical realization of quantitative phase imaging (QPI) on standard microscope platforms. The QPICWD is a compact stand-alone camera which measures sample induced phase delay under low-coherence quasi-monochromatic illumination by examining the deformation of the speckle intensity pattern. By interpreting the speckle deformation with an ensemble average of the geometric flow, we can obtain the high-resolution distortion field by solving the transport of intensity equation (TIE). Since the phase measured by TIE is the generalized phase of the partially coherent image, instead of the phase of the measured object, we analyze the effect of illumination coherence and imaging numerical aperture (NA) on the accuracy of phase retrieval, revealing that the phase of the object can be reliably retrieved when the coherence parameter (the ratio of illumination NA to objective NA) of the Kohler illumination is between 0.3 and 0.5. We present several applications for the new design including nondestructive optical testing of microlens array with nanometric thickness and imaging of fixed and live HeLa cells. Since the designed QPI camera does not require any modification of the widely available bright-field microscope or additional accessories for its use, it is expected to be adopted by the broader biology and medical community.

Compensation of Frequency Drift in Frequency-Sweep Polarization-Modulation Ranging System

In frequency-sweep polarization-modulation ranging, distance is determined by the frequency of modulated waves and the corresponding wavelength multiple when emitted and returned waves are in phase. However, measurement of the frequency and the wavelength multiple is affected by thermally induced phase delay of the polarized wave. In this article we systematically discuss the principle of the ranging method and analyze the influences of thermally induced phase delay. New approaches to measurement are proposed to eliminate the impact on frequency and the wavelength multiple. Theoretical analysis and experimental results proved the efficiency and applicability of the methods.

ER stress activation impairs the expression of circadian clock and clock-controlled genes in NIH3T3 cells via an ATF4-dependent mechanism

Endoplasmic reticulum (ER) stress and circadian clockwork signaling pathways mutually regulate various cellular functions, but the details regarding the cross-talk between these pathways in mammalian cells are unclear. In this study, whether perturbation of ER stress signaling affects the cellular circadian clockwork and transcription of clock-controlled genes was investigated in NIH3T3 mouse fibroblasts. An NIH3T3 cell model stably expressing luciferase (Luc) under the control of the Bmal1 clock gene promoter was established using a lentiviral system. Then, Luc activity was monitored in real-time to detect Bmal1-Luc oscillations. The ER stress activators thapsigargin (Tg) and tunicamycin (Tm) markedly reduced Bmal1-Luc oscillation amplitudes and induced phase delay shifts in NIH3T3 cells. Treatment with Tg/Tm activated ER stress signaling by upregulating GRP78, CHOP, ATF6, and ATF4 and simultaneously significantly decreased BMAL1 protein levels and inhibited the transcription of circadian clock (Bmal1, Per2, Nr1d1, and Dbp) and clock-controlled (Scad1, Fgf7, and Arnt) genes. 4-Phenylbutyric acid, an ER stress inhibitor, alleviated the transcriptional repression of the circadian clock genes and partially restored Bmal1-Luc oscillation amplitudes in Tg- or Tm-treated NIH3T3 cells. More importantly, knock-down of ATF4, but not ATF6, in Tg-treated NIH3T3 cells partially rescued Bmal1-Luc oscillation amplitudes and mRNA expression of the four circadian clock genes. Taken together, our study demonstrates that ER stress activation inhibits the transcription of circadian clock and clock-controlled genes via an ATF4-dependent mechanism.

Functional Significance of the Excitatory Effects of GABA in the Suprachiasmatic Nucleus

Over 90% of neurons within the suprachiasmatic nucleus (SCN) express γ-aminobutyric acid (GABA). Although GABA is primarily an inhibitory neurotransmitter, in vitro studies suggest that the activation of GABAA receptors (GABAAR) elicits excitation in the adult SCN. The ratio of excitatory to inhibitory responses to GABA depends on the balance of chloride influx by Na+-K+-Cl- cotransporter 1 (NKCC1) and chloride efflux by K+-Cl- cotransporters (KCCs). Excitatory responses to GABA can be blocked by inhibition of the inward chloride cotransporter, NKCC1, with the loop diuretic bumetanide. Here we investigated the role of NKCC1 activity in phase shifting the circadian pacemaker in response to photic and nonphotic signals in male Syrian hamsters housed in constant darkness. In the early subjective night (CT 13.5), injection of bumetanide into the SCN reduced light-induced phase delays. However, during the late subjective night (CT 19), bumetanide administration did not alter light-induced phase advances. Injection of bumetanide during the subjective day (CT 6) did not alter the phase of free-running circadian rhythms but attenuated phase advances induced by injection of the GABAAR agonist muscimol into the SCN. These data support the hypothesis that the excitatory effects of endogenously released GABA contribute to the ability of light to induce phase delays, thereby contributing to the most important function of the circadian system, its entrainment with the day-night cycle. Further, the finding that bumetanide inhibits the phase-advancing effects of muscimol during the subjective day supports the hypothesis that the excitatory responses to GABA also contribute to the ability of nonphotic stimuli to phase shift the circadian pacemaker.

Sleep Deprivation and Caffeine Treatment Potentiate Photic Resetting of the Master Circadian Clock in a Diurnal Rodent.

Circadian rhythms in nocturnal and diurnal mammals are primarily synchronized to local time by the light/dark cycle. However, nonphotic factors, such as behavioral arousal and metabolic cues, can also phase shift the master clock in the suprachiasmatic nuclei (SCNs) and/or reduce the synchronizing effects of light in nocturnal rodents. In diurnal rodents, the role of arousal or insufficient sleep in these functions is still poorly understood. In the present study, diurnal Sudanian grass rats, Arvicanthis ansorgei, were aroused at night by sleep deprivation (gentle handling) or caffeine treatment that both prevented sleep. Phase shifts of locomotor activity were analyzed in grass rats transferred from a light/dark cycle to constant darkness and aroused in early night or late night. Early night, but not late night, sleep deprivation induced a significant phase shift. Caffeine on its own induced no phase shifts. Both sleep deprivation and caffeine treatment potentiated light-induced phase delays and phase advances in response to a 30 min light pulse, respectively. Sleep deprivation in early night, but not late night, potentiated light-induced c-Fos expression in the ventral SCN. Caffeine treatment in midnight triggered c-Fos expression in dorsal SCN. Both sleep deprivation and caffeine treatment potentiated light-induced c-Fos expression in calbindin-containing cells of the ventral SCN in early and late night. These findings indicate that, in contrast to nocturnal rodents, behavioral arousal induced either by sleep deprivation or caffeine during the sleeping period potentiates light resetting of the master circadian clock in diurnal rodents, and activation of calbindin-containing suprachiasmatic cells may be involved in this effect.SIGNIFICANCE STATEMENT Arousing stimuli have the ability to regulate circadian rhythms in mammals. Behavioral arousal in the sleeping period phase shifts the master clock in the suprachiasmatic nuclei and/or slows down the photic entrainment in nocturnal animals. How these stimuli act in diurnal species remains to be established. Our study in a diurnal rodent, the Grass rat, indicates that sleep deprivation in the early rest period induces phase delays of circadian locomotor activity rhythm. Contrary to nocturnal rodents, both sleep deprivation and caffeine-induced arousal potentiate the photic entrainment in a diurnal rodent. Such enhanced light-induced circadian responses could be relevant for developing chronotherapeutic strategies.

White-light quantitative phase imaging unit

We introduce the white-light quantitative phase imaging unit (WQPIU) as a practical realization of quantitative phase imaging (QPI) on standard microscope platforms. The WQPIU is a compact stand-alone unit which measures sample induced phase delay under white-light illumination. It does not require any modification of the microscope or additional accessories for its use. The principle of the WQPIU based on lateral shearing interferometry and phase shifting interferometry provides a cost-effective and user-friendly use of QPI. The validity and capacity of the presented method are demonstrated by measuring quantitative phase images of polystyrene beads, human red blood cells, HeLa cells and mouse white blood cells. With speckle-free imaging capability due to the use of white-light illumination, the WQPIU is expected to expand the scope of QPI in biological sciences as a powerful but simple imaging tool.

Sustained activation of GABAA receptors in the suprachiasmatic nucleus mediates light-induced phase delays of the circadian clock: a novel function of ionotropic receptors.

The suprachiasmatic nucleus (SCN) contains a circadian clock that generates endogenous rhythmicity and entrains that rhythmicity with the day-night cycle. The neurochemical events that transduce photic input within the SCN and mediate entrainment by resetting the molecular clock have yet to be defined. Because GABA is contained in nearly all SCN neurons we tested the hypothesis that GABA serves as this signal in studies employing Syrian hamsters (Mesocricetus auratus). Activation of GABAA receptors was found to be necessary and sufficient for light to induce phase delays of the clock. Remarkably, the sustained activation of GABAA receptors for more than three consecutive hours was necessary to phase-delay the clock. The duration of GABAA receptor activation required to induce phase delays would not have been predicted by either the prevalent theory of circadian entrainment or by expectations regarding the duration of ionotropic receptor activation necessary to produce functional responses. Taken together, these data identify a novel neurochemical mechanism essential for phase-delaying the 'master' circadian clock within the SCN as well as identifying an unprecedented action of an amino acid neurotransmitter involving the sustained activation of ionotropic receptors.

Bright and dark plasmon resonances of nanoplasmonic antennas evanescently coupled with a silicon nitride waveguide

In this work we investigate numerically and experimentally the resonance wavelength tuning of different nanoplasmonic antennas excited through the evanescent field of a single mode silicon nitride waveguide and study their interaction with this excitation field. Experimental interaction efficiencies up to 19% are reported and it is shown that the waveguide geometry can be tuned in order to optimize this interaction. Apart from the excitation of bright plasmon modes, an efficient coupling between the evanescent field and a dark plasmonic resonance is experimentally demonstrated and theoretically explained as a result of the propagation induced phase delay.

Selective pharmacological blockade of the 5-HT7 receptor attenuates light and 8-OH-DPAT induced phase shifts of mouse circadian wheel running activity.

Recent reports have illustrated a reciprocal relationship between circadian rhythm disruption and mood disorders. The 5-HT7 receptor may provide a crucial link between the two sides of this equation since the receptor plays a critical role in sleep, depression, and circadian rhythm regulation. To further define the role of the 5-HT7 receptor as a potential pharmacotherapy to correct circadian rhythm disruptions, the current study utilized the selective 5-HT7 antagonist JNJ-18038683 (10 mg/kg) in three different circadian paradigms. While JNJ-18038683 was ineffective at phase shifting the onset of wheel running activity in mice when administered at different circadian time (CT) points across the circadian cycle, pretreatment with JNJ-18038683 blocked non-photic phase advance (CT6) induced by the 5-HT1A/7 receptor agonist 8-OH-DPAT (3 mg/kg). Since light induced phase shifts in mammals are partially mediated via the modulation of the serotonergic system, we determined if JNJ-18038683 altered phase shifts induced by a light pulse at times known to phase delay (CT15) or advance (CT22) wheel running activity in free running mice. Light exposure resulted in a robust shift in the onset of activity in vehicle treated animals at both times tested. Administration of JNJ-18038683 significantly attenuated the light induced phase delay and completely blocked the phase advance. The current study demonstrates that pharmacological blockade of the 5-HT7 receptor by JNJ-18038683 blunts both non-photic and photic phase shifts of circadian wheel running activity in mice. These findings highlight the importance of the 5-HT7 receptor in modulating circadian rhythms. Due to the opposite modulating effects of light resetting between diurnal and nocturnal species, pharmacotherapy targeting the 5-HT7 receptor in conjunction with bright light therapy may prove therapeutically beneficial by correcting the desynchronization of internal rhythms observed in depressed individuals.

Suprachiasmatic Astrocytes Modulate the Circadian Clock in Response to TNF-α

The immune and the circadian systems interact in a bidirectional fashion. The master circadian oscillator, located in the suprachiasmatic nuclei (SCN) of the hypothalamus, responds to peripheral and local immune stimuli, such as proinflammatory cytokines and bacterial endotoxin. Astrocytes exert several immune functions in the CNS, and there is growing evidence that points toward a role of these cells in the regulation of circadian rhythms. The aim of this work was to assess the response of SCN astrocytes to immune stimuli, particularly to the proinflammatory cytokine TNF-α. TNF-α applied to cultures of SCN astrocytes from Per2(luc) knockin mice altered both the phase and amplitude of PER2 expression rhythms, in a phase-dependent manner. Furthermore, conditioned media from SCN astrocyte cultures transiently challenged with TNF-α induced an increase in Per1 expression in NIH 3T3 cells, which was blocked by TNF-α antagonism. In addition, these conditioned media could induce phase shifts in SCN PER2 rhythms and, when administered intracerebroventricularly, induced phase delays in behavioral circadian rhythms and SCN activation in control mice, but not in TNFR-1 mutants. In summary, our results show that TNF-α modulates the molecular clock of SCN astrocytes in vitro, and also that, in response to this molecule, SCN astrocytes can modulate clock gene expression in other cells and tissues, and induce phase shifts in a circadian behavioral output in vivo. These findings suggest a role for astroglial cells in the alteration of circadian timing by immune activation.

Role of Proinflammatory Cytokines on Lipopolysaccharide-Induced Phase Shifts in Locomotor Activity Circadian Rhythm

We previously reported that early night peripheral bacterial lipopolysaccharide (LPS) injection produces phase delays in the circadian rhythm of locomotor activity in mice. We now assess the effects of proinflammatory cytokines on circadian physiology, including their role in LPS-induced phase shifts. First, we investigated whether differential systemic induction of classic proinflammatory cytokines could explain the time-specific behavioral effects of peripheral LPS. Induction levels for plasma interleukin (IL)-1α, IL-1β, IL-6, or tumor necrosis factor (TNF)-α did not differ between animals receiving a LPS challenge in the early day or early night. We next tested the in vivo effects of central proinflammatory cytokines on circadian physiology. We found that intracerebroventricular (i.c.v.) delivery of TNF-α or interleukin IL-1β induced phase delays on wheel-running activity rhythms. Furthermore, we analyzed if these cytokines mediate the LPS-induced phase shifts and found that i.c.v. administration of soluble TNF-α receptor (but not an IL-1β antagonistic) prior to LPS stimulation inhibited the phase delays. Our work suggests that the suprachiasmatic nucleus (SCN) responds to central proinflammatory cytokines in vivo, producing phase shifts in locomotor activity rhythms. Moreover, we show that the LPS-induced phase delays are mediated through the action of TNF-α at the central level, and that systemic induction of proinflammatory cytokines might be necessary, but not sufficient, for this behavioral outcome. (Author correspondence: dgolombek@unq.edu.ar)

Human responses to bright light of different durations

Light exposure in the early night induces phase delays of the circadian rhythm in melatonin in humans. Previous studies have investigated the effect of timing, intensity, wavelength, history and pattern of light stimuli on the human circadian timing system. We present results from a study of the duration–response relationship to phase-delaying bright light. Thirty-nine young healthy participants (16 female; 22.18±3.62 years) completed a 9-day inpatient study. Following three baseline days, participants underwent an initial circadian phase assessment procedure in dim light (<3 lux), and were then randomized for exposure to a bright light pulse (∼10,000 lux) of 0.2 h, 1.0 h, 2.5 h or 4.0 h duration during a 4.5 h controlled-posture episode centred in a 16 h wake episode. After another 8 h sleep episode, participants completed a second circadian phase assessment. Phase shifts were calculated from the difference in the clock time of the dim light melatonin onset (DLMO) between the initial and final phase assessments. Exposure to varying durations of bright light reset the circadian pacemaker in a dose-dependent, non-linear manner. Per minute of exposure, the 0.2 h duration was over 5 times more effective at phase delaying the circadian pacemaker (1.07±0.36 h) as compared with the 4.0 h duration (2.65±0.24 h). Acute melatonin suppression and subjective sleepiness also had a dose-dependent response to light exposure duration. These results provide strong evidence for a non-linear resetting response of the human circadian pacemaker to light duration.

Anti‐Phase Expression of the BK Channel (Kcnma1) Alters Circadian Locomotor Activity in Mice

Circadian rhythmicity, driven by the suprachiasmatic nucleus (SCN) of the hypothalamus, is essential to the survival or fitness of all organisms. The BK Ca2+ and voltage‐activated K+ channel is expressed in a circadian manner in the SCN, peaking at night, and genetic deletion of the BK channel (Kcnma1−/−) disrupts circadian behavioral rhythms. To determine whether the nighttime‐phased expression pattern was essential for normal circadian rhythmicity, we analyzed wheel running activity of transgenic mice that expressed the BK channel opposite to its normal phase in the SCN (Per1:BKR207Q ). We predicted this alteration of BK phase by expression of one or two copies (1C, 2C) of the Per1:BKR207Q transgene, or one copy of Per1:BKR207Q with one deleted endogenous copy of Kcnma1 (1C; BK het) would disrupt circadian wheel rhythms. In contrast, 1C, 2C, and 1C; BK het mice all had relatively normal baseline circadian rhythms. However, 2C mice had a longer active interval compared to WT, suggesting that nighttime‐phased expression of the BK channel is important for restricting locomotor activity. Additionally, 2C mice had a larger light‐pulse induced phase delays, suggesting these mice were more sensitive to phase‐shifting stimuli than WT. A better understanding of the neural encoding of circadian rhythms may impact the treatment of sleep and other disorders of circadian rhythmicity.This work was funded by an APS summer research fellowship to (B.N.W.) and NSF (IOS‐0956237) and NHLBI (R01HL102758) to A.L.M.

Light-Induced Changes of the Circadian Clock of Humans: Increasing Duration is More Effective than Increasing Light Intensity

To evaluate the effect of increasing the intensity and/or duration of exposure on light-induced changes in the timing of the circadian clock of humans. Multifactorial randomized controlled trial, between and within subject design General Clinical Research Center (GCRC) of an academic medical center 56 healthy young subjects (20-40 years of age) Research subjects were admitted for 2 independent stays of 4 nights/3 days for treatment with bright or dim-light (randomized order) at a time known to induce phase delays in circadian timing. The intensity and duration of the bright light were determined by random assignment to one of 9 treatment conditions (duration of 1, 2, or 3 hours at 2000, 4000, or 8000 lux). Treatment-induced changes in the dim light melatonin onset (DLMO) and dim light melatonin offset (DLMOff) were measured from blood samples collected every 20-30 min throughout baseline and post-treatment nights. Comparison by multi-factor analysis of variance (ANOVA) of light-induced changes in the time of the circadian melatonin rhythm for the 9 conditions revealed that changing the duration of the light exposure from 1 to 3 h increased the magnitude of light-induced delays. In contrast, increasing from moderate (2,000 lux) to high (8,000 lux) intensity light did not alter the magnitude of phase delays of the circadian melatonin rhythm. Results from the present study suggest that for phototherapy of circadian rhythm sleep disorders in humans, a longer period of moderate intensity light may be more effective than a shorter exposure period of high intensity light.

An NMDA antagonist inhibits light but not GRP-induced phase shifts when administered after the phase-shifting stimulus

Brief light pulses or microinjection of gastrin-releasing peptide (GRP) into the third ventricle or near the suprachiasmatic nucleus (SCN) induce phase shifts of circadian rhythms during the subjective night. It has previously been reported that these effects are strongly influenced by the activation of N-methyl- d-aspartate (NMDA) receptors and the availability of glutamate. We hypothesized that the photic signaling pathway in the SCN was dependent on glutamate neurotransmission even after the completion of a photic stimulus. Adult male Syrian hamsters equipped with a surgically implanted guide cannula aimed at the SCN region were housed in constant darkness until stable free-running rhythms of wheel-running activity were apparent. Light pulses administered in the early night induced phase delays of circadian rhythms which were attenuated by the co-administration of (±)-2-amino-5-phosphonopentanoic acid (AP5), an NMDA antagonist. Microinjection of AP5 also inhibited light-induced shifts, to a lesser extent, immediately after and 15 min after, but not 30 min after the light pulse. A second experiment was designed to test whether AP5 would be able to attenuate GRP-induced shifts 15 min following microinjection of GRP. Phase shifts of animals that received microinjection of AP5 15 min after the administration of GRP were not different from those that received microinjection of GRP and vehicle. These data suggest that glutamate signaling remains necessary for a full photic response in the SCN even after the termination of the photic signal, but that this dependency ends once GRP-dependent signaling is complete.

Paying the circadian toll: The circadian response to LPS injection is dependent on the Toll-like receptor 4

Systemic low doses of the endotoxin lipopolysaccharide (LPS) administered at CT15 (circadian time 12 corresponds to locomotor activity onset) induce phase delays of locomotor activity rhythms in mice. To evaluate if this effect was mediated by the Toll-like receptor 4 (TLR4), our present aim was to characterize the circadian behavior and LPS-induced circadian response of TLR4 (LPS receptor)-deficient mice (in C57bl/10 and C3H backgrounds). In mutants, we observed a free-running period and a light-induced phase delay similar to the one observed in their corresponding wild-type (WT) littermates. The LPS-induced phase delay, wheel running inhibition and c-Fos/Per-1 immunoreactivity in the paraventricular nuclei observed in WT mice was absent or significantly decreased in the TLR4-deficient mice. In conclusion, we show that LPS-induced circadian responses are mediated by TLR4.

EFFECTS OF LIGHT ON THE CIRCADIAN ACTIVITY RHYTHM OF DJUNGARIAN HAMSTERS (PHODOPUS SUNGORUS) WITH DELAYED ACTIVITY ONSET

A number of Djungarian hamsters (Phodopus sungorus) of our institute show activity patterns that seem incompatible with proper adjustment to a periodic environment. The activity onset of those animals is continuously delayed, whereas the activity offset is stably coupled to “lights-on”, leading to compression of activity time. A series of experiments was conducted to evaluate the possible causes of the deteriorated ability of DAO (delayed activity onset) hamsters to synchronize. Thus, we investigated the properties of the endogenous circadian rhythm plus parametric and non-parametric light effects on hamsters of DAO and Wild type (WT) phenotypes. Free-running rhythms were studied in constant darkness (DD) or constant light (LL) of different intensities (1, 10, 100 lux). To investigate photic phase responses, hamsters were kept in DD and exposed to light pulses (100 lux, 15 min), at circadian time (CT) CT14 and CT22. Differences were verified statistically by ANOVA. Light intensity exerted significant effect on the free-running period (τ). In DD, τ was significantly longer in DAO than WT hamsters. With increasing light intensity, τ lengthened in both phenotypes, though not at a similar rate. In 10 and 100 lux LL, however, τ did not differ between the two phenotypes. The robustness of the circadian activity rhythm was highest in DD and decreased in LL. No differences between phenotypes were noted. The percentage of arrhythmic animals was low in DD, but remarkably high in LL, and always higher in WT hamsters. The total amount of activity/day was highest in DD; DAO hamsters were less active than WT hamsters under each lighting condition. Light pulses induced phase delays when applied at CT14 and phase advances at CT22, with advances being stronger than delays. Also at CT14, the response of the activity onset was stronger than the activity offset. The opposite was observed at CT22. At CT14, the phase response did not differ between the phenotypes. However, at CT22 the phase advance was significantly weaker in DAO than WT hamsters despite their longer τ. The results provide further evidence that the distinct activity pattern of DAO hamsters is due to an altered interaction between the circadian clock and photic zeitgeber. (Author correspondence: weinert@zoologie.uni-halle.de)