Effects of prolonged exposure to darkness on circadian photic responsiveness in the mouse

Circadian rhythms in mammals are generated by an endogenous pacemaker but are modulated by environmental cycles, principally the alternation of light and darkness. Although much is known about nonparametric effects of light on the circadian system, little is known about other effects of photic stimulation. In the present study, which consists of a series of five experiments in mice, various manipulations of photic stimulation were used to dissect the mechanisms responsible for a variation in the magnitude of light-induced phase-shifts that results from prolonged exposure to darkness. The results confirmed previous observations that prolonged exposure to darkness causes an increase in the magnitude of phase shifts (both phase advances and phase delays) evoked by discrete light pulses. The results also indicated that the increase in responsiveness results from the lack of exposure to light per se and not from collateral effects of exposure to constant darkness such as the lack of previous entrainment. The lack of exposure to light causes the circadian system to undergo a process of dark adaptation similar to dark adaptation in the visual system but with a much slower temporal course. The results suggest that circadian dark adaptation may take place at the retinal level, but it is not clear whether it involves a change in the sensitivity or maximal responsiveness of the system.     

Effects of Prolonged Exposure to Darkness on Circadian Photic Responsiveness in the Mouse

Circadian rhythms in mammals are generated by an endogenous pacemaker but are modulated by environmental cycles, principally the alternation of light and darkness. Although much is known about nonparametric effects of light on the circadian system, little is known about other effects of photic stimulation. In the present study, which consists of a series of five experiments in mice, various manipulations of photic stimulation were used to dissect the mechanisms responsible for a variation in the magnitude of light-induced phase-shifts that results from prolonged exposure to darkness. The results confirmed previous observations that prolonged exposure to darkness causes an increase in the magnitude of phase shifts (both phase advances and phase delays) evoked by discrete light pulses. The results also indicated that the increase in responsiveness results from the lack of exposure to light per se and not from collateral effects of exposure to constant darkness such as the lack of previous entrainment. The lack of exposure to light causes the circadian system to undergo a process of dark adaptation similar to dark adaptation in the visual system but with a much slower temporal course. The results suggest that circadian dark adaptation may take place at the retinal level, but it is not clear whether it involves a change in the sensitivity or maximal responsiveness of the system.     

The effects of pinealectomy and blinding on the circadian locomotor activity rhythm in the Japanese newt,Cynops pyrrhogaster

We examined the effects of pinealectomy and blinding (bilateral ocular enucleation) on the circadian locomotor activity rhythm in the Japanese newt, Cynops pyrrhogaster. The pinealectomized newts were entrained to a light-dark cycle of 12 h light and 12 h darkness. After transfer to constant darkness they showed residual rhythmicity for at least several days which was gradually disrupted in prolonged constant darkness. Blinded newts were also entrained to a 12 h light/12 h dark cycle. In subsequent constant darkness they showed free-running rhythms of locomotor activity. However, the freerunning periods noticeably increased compared with those observed in the previous period of constant darkness before blinding. In blinded newts entrained to the light/dark cycle the activity rhythms were gradually disrupted after pinealectomy even in the presence of the light/dark cycle. These results suggest that both the pineal and the eyes are involved in the newt's circadian system, and also suggest that the pineal of the newt acts as an extraretinal photoreceptor which mediates the entrainment of the locomotor activity rhythm.Abbreviations circadian period - DD constant darkness - LD cycle, light-dark cycle - LD 12:12 light-dark cycle of 12 h light and 12 h darkness     

Circadian Intraocular Pressure Rhythms in Athletic Horses under Different Lighting Regime

The present study was undertaken to investigate the existence of intraocular pressure (IOP) rhythms in athletic thoroughbred horses maintained under a 24 h cycle of light and darkness (LD) or under constant light (LL) or constant dark (DD) conditions. We identified an IOP circadian rhythm that is entrained to the 24 h LD cycle. IOP was low during the dark phase and high during the light phase, with a peak at the end of the light phase (ZT10). The circadian rhythm of IOP persisted in DD (with a peak at CT9.5), demonstrating an endogenous component in IOP rhythm. As previously shown in other mammalian species, horse IOP circadian rhythmicity was abolished in LL. Because tonometry is performed in horses for the diagnosis of ophthalmologic diseases, such as glaucoma or anterior uveitis, the daily variation in IOP must be taken into account in clinical practice to properly time tests and to interpret clinical findings.     

Effect of melatonin agonists and antagonists on horizontal cell spinule formation and dopamine release in a fish retina

The crucian carp retina was used to study the effects of the melatonin antagonist p697 (N-pentanoyl 2-benzyltryptamine) and the melatonin agonists [+]- and [-]-AMMTC (N-acetyl-4-aminomethyl-6-methoxy-9-methyl-1,2,3,4-tetrahydrocarbazol e) on horizontal cell spinule formation, an indicator of the state of retinal adaptation. DH97 was capable of both counteracting dark-adaptive spinule degradation and inducing light-adaptive spinule formation at the beginning of the dark phase. Addition of dopamine receptor blockers opposed the action of DH97 on spinules, with SCH 23930, a D1 dopamine receptor antagonist, being more effective than the D2 receptor antagonist sulpiride. DH97 induced a twofold increase in dopamine release. We conclude that melatonin acts as a dark signal within the teleost retina by inhibiting the dopaminergic system. In accordance with this, both enantiomers of AMMTC prevented light-induced spinule formation, and reduced dopamine release to below dark-adaptive baseline levels. We suggest that the suppression of spinule formation by AMMTC may be due to either a direct inhibitory interaction between the melatonin agonist and horizontal cell dopamine D1 receptors, or an inhibitory effect on the activity of the dopamine-releasing interplexiform cells.     

Endogenous patterns of photomechanical movements in teleosts and their relation to activity rhythms

Summary The retinal rods, cones and epithelial pigment of most lower vertebrates display rhythmic photomechanical (retinomotor) migrations in response to changes in ambient lighting conditions. This study examines the extent of these migrations in the absence of the daily changes in illumination (constant darkness and constant light) in three species of teleosts. Salmo trutta, a crepuscularly active fish, showed two peaks of light adaptation occurring around dawn and dusk when kept in constant darkness. Tinca tinca, a nocturnal species, also showed an endogenous rhythm during extended periods of darkness, but, unlike Salmo trutta, it was light-adapted throughout what would normally have been day. At the maximal extent of migration under conditions of continual darkness, the pigment migrated 59% as much as it did during a normal light/dark cycle. Nannacara anomala, a tropical diurnally active species, showed a similar but more pronounced rhythm than Tinea tinea for all 3 days of experimental darkness, behaving essentially identically to fish exposed to a light/dark cycle. Nannacara anomala also displayed a weak rhythm when kept in constant light.It is concluded from these and previous results that the pattern of endogenous photomechanical movement depends both on the activity pattern of a species and on the constancy of the lighting conditions to which it has been exposed during its lifetime.     

Melatonin accelerates reentrainment of the circadian rhythm of its own production after an 8-h advance of the light-dark cycle

Summary The rhythm in melatonin production in the rat is driven by a circadian rhythm in the pineal N-acetyltransferase (NAT) activity. Rats adapted to an artificial lighting regime of 12 h of light and 12 h of darkness per day were exposed to an 8-h advance of the light-dark regime accomplished by the shortening of one dark period; the effect of melatonin, triazolam and fluoxetine, together with 5-hydroxytryptophan, on the reentrainment of the NAT rhythm was studied.In control rats, the NAT rhythm was abolished during the first 3 cycles following the advance shift. It reappeared during the 4th cycle; however, the phase relationship between the evening rise in activity and the morning decline was still compressed.Melatonin accelerated the NAT rhythm reentrainment. In rats treated chronically with melatonin at the new dark onset, the rhythm had already reappeared during the 3rd cycle, in the middle of the advanced night, and during the 4th cycle, the phase relationship between the evening onset and the morning decline of the NAT activity was the same as before the advance shift. In rats treated chronically with melatonin at the old dark onset or in those treated with melatonin 8 h, 5 h and 2 h after the new dark onset during the 1st, 2nd and 3rd cycle, respectively, following the advance shift, the NAT rhythm reappeared during the 3rd cycle as well but in the last third of the advanced night only.Neither triazolam nor fluoxetine together with 5-hydroxytryptophan administered around the new dark onset facilitated NAT rhythm reentrainment after the 8-h advance of the light-dark cycle.Abbreviations NAT N-acetyltransferase - LD cycle light-dark cycle - CT circadian time - LD xy light dark cycle comprising x h of light and y h of darkness     

Ecdysteroids influence the circadian system timing ecdysis in the insectRhodnius prolixus (Hemiptera)

Summary 20-hydroxyecdysone (20HE) injections induced transient delays in the time of ecdysis inRhodnius prolixus reared in L/D cycles. Sustained phase delays in the ecdysis rhythm were revealed by transfer to constant dark during the scotophase following 20HE injection. The magnitude of the phase delays depended on the time in the L/D cycle at which 20HE was injected with major delays occurring at times when the endogenous titre is declining. Therefore the increases and decreases in the endogenous titre which are themselves timed in a circadian fashion may be involved in phase setting the ecdysis rhythm to the environmental cycle. Populations maintained in LL which are arrhythmic with respect to both ecdysteroid titres and ecdysis, can be induced to display gated ecdysis by injection of either 20HE or antiserum to ecdysteroids. Multiple injections of 20HE or antiserum are capable of inducing an ecdysis rhythm whose period (22.3 h) and gate location are very similar to that produced by altering the environmental cycle. Therefore manipulations of the endogenous titre of ecdysteroids can mimic the effects of L/D cycles on the timing of ecdysis. Ecdysis inRhodnius may therefore be timed at least partially as a result of circadian timing of the ecdysteroid titre.Abbreviations AZT Arbitrary Zeitgeber Time - DD constant darkness - LL constant light - L/D 24 h light dark cycle - 12L/12D 12 h of light 12 h of dark - 20HE 20-hydroxyecdysone     

Presence of circadian rhythms in the locomotor activity of a cave-dwelling millipede Glyphiulus cavernicolus sulu (Cambalidae, Spirostreptida)

The locomotor activity of the millipede Glyphiulus cavernicolus (Spirostreptida), which occupies the deeper recesses of a cave, was monitored in light-dark (LD) cycles (12h light and 12h darkness), constant darkness (DD), and constant light (LL) conditions. These millipedes live inside the cave and are apparently never exposed to any periodic factors of the environment such as light-dark, temperature, and humidity cycles. The activity of a considerable fraction of these millipedes was found to show circadian rhythm, which entrained to a 12:12 LD cycle with maximum activity during the dark phase of the LD cycle. Under constant darkness (DD), 56.5% of the millipedes (n = 23) showed circadian rhythms, with average free-running period of 25.7h ± 3.3h (mean ± SD, range 22.3h to 35.0h). The remaining 43.5% of the millipedes, however, did not show any clear-cut rhythm. Under DD conditions following an exposure to LD cycles, 66.7% (n = 9) showed faint circadian rhythm, with average free-running period of 24.0h ± 0.8h (mean ± SD, range 22.9h to 25.2h). Under constant light (LL) conditions, only 2 millipedes of 11 showed free-running rhythms, with average period length of 33.3h ± 1.3h. The results suggest that these cave-dwelling millipedes still possess the capacity to measure time and respond to light and dark situations. (Chronobiology International, 17(6), 757-765, 2000)     

Feeding rhythms in constant light and constant darkness: the role of the eyes and the effect of melatonin infusion

Exposure to constant light abolishes circadian behavioral rhythms of locomotion and feeding as well as circulating melatonin rhythms in pigeons (Columba livia). To determine if feeding rhythmicity could be maintained in pigeons exposed to constant light, periodic infusions (10h/day) of melatonin were administered to pinealectomized and bilaterally retinectomized/pinealectomized pigeons under conditions of both constant darkness and constant light. The infusions were sufficient to entrain rhythmicity in pinealectomized pigeons in constant darkness and to restore and maintain rhythmicity in bilaterally retinectomized/pinealectomized pigeons in constant darkness. On subsequent exposure to constant light, rhythmicity remained phase locked to the melatonin infusions in bilaterally retinectomized/pinealectomized pigeons but was abolished in sighted pinealectomized birds. These results suggest that while endogenous melatonin rhythms are both necessary and sufficient to maintain behavioral rhythms in DD, their effect can be overridden by constant light but only if perceived by the eyes. Thus, constant light may abolish behavioral rhythmicity in intact pigeons (and perhaps in other species) by a mechanism other than suppression of endogenous melatonin rhythmicity. Such a mechanism might involve direct stimulation of locomotor or feeding activity by retinally perceived (but not by extra-retinally perceived) light, or alternatively by suppression of a hypothalamic oscillator that receives its major light input from the retinae.Abbreviations PX pinealectomized - EX bilaterally enucleated - LD light:dark cycle - LL constant light - DD constant darkness - DD_b constant darkness before exposure to constant light - DD_a constant darkness after exposure to constant light     

Stem extension rate in light-grown plants. Evidence for an endogenous circadian rhythm in Chenopodium rubrum

Stem extension in light-grown plants of Chenopodium rubrum L. ecotype selection 184 (50°10'N; 150°35'W) was recorded continuously for periods up to one week at constant temperature. Stem extension rate measurements were made with linear voltage-displacement transducer devices. At the beginning of experiments, the 3rd intenode above the cotyledons was about 5 mm long. Stem extension rate exhibited a rhythmic behaviour in continuous white light (20 W m⁻²), and in continuous darkness with a period of approximately 23 h. In continuous darkness, the amplitude of the rhythm damped out very quickly after 24 h and a second peak was just measurable. The mean value of the stem extension rate was dependent on the light fluence before the experiments. This overt rhythm, which could be observed at the individual plant or even internode level, exhibited the characteristics of an endogenous circadian rhythm. There was no correlation of the peak time to local time. The peak time was determined by the time of transfer from dark to light for dark periods equal to or longer than 8 h, and the phase was shifted by the time of transfer from light to dark at the proper phase of a pre-existing rhythm.     

Diurnal Fluctuations in Ethylene Formation in Chenopodium rubrum

Ethylene formation was studied in 5- to 6-d-old Chenopodium rubrum seedlings under the following light regimes: continuous light (CL), continuous darkness (CD), and alternating light/darkness (12 h of each). No significant regular oscillations in ethylene formation were found in either the CL or CD groups. In the light/dark regime, pronounced diurnal fluctuations in ethylene formation were observed. Activity of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase was transiently increased on transfer from light to dark and vice versa. In CL, ACC oxidase activity did not change significantly, whereas in CD, it decreased continuously after the initial increase. The in vivo levels of ACC and N-malonyl-ACC (MACC) were constant for the first few hours of darkness, then decreased dramatically, but increased again in the light. In constant darkness, the level of ACC displayed endogenous rhythm, with minimum values at h 12 and 44, and a maximum value at h 32 to 36. The level of MACC in both shoots and roots decreased in the CD group until h 12, and then remained constant until h 30 before decreasing continuously. We conclude that the photoperiodic regime affects both ACC and MACC levels, as well as the conversion of ACC to ethylene. Correlation of the described changes in ethylene formation to photoperiodic flower induction is discussed.     

Influence of experimental illumination and seasonal variation on crossbreeding mating in the snail Biomphalaria glabrata

The crossbreeding activities of the Schistosoma mansoni vector snail Biomphalaria glabrata were counted in a laboratory aquarium throughout the year under two regimes of 12h light: 12h dark from 7 A. M. to 10 P. M. Mating increased significantly in Autumn and Winter and just missed a significant inverse correlation with temperature and a direct one with locomotion. Other similar experiments were carried out to compare mating under various illumination conditions in complete daily cycle measurements. Mating counts decreased under the regimes which submitted snail to a total exposure of 12h light and 12h dark during a daily cycle in the following sequence: 12h light:12h dark alternating hourly with light gradient, 12h light:12h dark, 1h light:1h dark and 12h dark:12h light. Under two constant illuminations, the mating scored less than under the previous conditions, except under 12h dark:12h light. Under darkness the mating count was lower than under light conditions. There was no way to differentiate the night and day rhythms of mating on different days in each regime, except for mating under 12h light:12h dark alternating with light gradient, constant dark and 12h dark:12h light conditions. Mating increased in certain light and temperature conditions, in which the intensities should have an optimum value.     

PRESENCE OF CIRCADIAN RHYTHMS IN THE LOCOMOTOR ACTIVITY OF A CAVE-DWELLING MILLIPEDE GLYPHIULUS CAVERNICOLUS SULU (CAMBALIDAE,SPIROSTREPTIDA)

The locomotor activity of the millipede Glyphiulus cavernicolus (Spirostreptida), which occupies the deeper recesses of a cave, was monitored in light-dark (LD) cycles (12h light and 12h darkness), constant darkness (DD), and constant light (LL) conditions. These millipedes live inside the cave and are apparently never exposed to any periodic factors of the environment such as light-dark, temperature, and humidity cycles. The activity of a considerable fraction of these millipedes was found to show circadian rhythm, which entrained to a 12:12 LD cycle with maximum activity during the dark phase of the LD cycle. Under constant darkness (DD), 56.5% of the millipedes (n = 23) showed circadian rhythms, with average free-running period of 25.7h ± 3.3h (mean ± SD, range 22.3h to 35.0h). The remaining 43.5% of the millipedes, however, did not show any clear-cut rhythm. Under DD conditions following an exposure to LD cycles, 66.7% (n = 9) showed faint circadian rhythm, with average free-running period of 24.0h ± 0.8h (mean ± SD, range 22.9h to 25.2h). Under constant light (LL) conditions, only 2 millipedes of 11 showed free-running rhythms, with average period length of 33.3h ± 1.3h. The results suggest that these cave-dwelling millipedes still possess the capacity to measure time and respond to light and dark situations. (Chronobiology International, 17(6), 757–765, 2000)     

Circadian control of spontaneous flight activity in the blowfly, Lucilia cuprina

ABSTRACT. Under laboratory light: dark cycles, the flight activity of adult Lucilia cuprina (Wied.) was low during darkness and uniformity high during light. This pattern persisted as a rhythm both in constant darkness and in constant light of intensity up to 1lx, with a period of approximately 22 h in each. Light pulses of 15 min at l00lx applied to the free-running rhythm in constant darkness generated phase shifts of up to 60°, 12-h light pulses of the same intensity generated maximal (180°) phase shifts. The phase response curves had shapes similar to those of a number of other insect rhythms. When exposed to light periods (70 lx) of greater than 12 h followed by constant darkness, the rhythm reinitiated at the light-dark transition from a constant phase equivalent to that at the time of the light-dark transition in the LD 12:12 cycle.     

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.     

ACTIVITY RHYTHMS OF WILD AND LABORATORY GOLDEN HAMSTERS (MESOCRICETUS AURATUS) UNDER ENTRAINED AND FREE-RUNNING CONDITIONS

The golden hamster (Mesocricetus auratus) is one of the most frequently used laboratory animals, particularly in chronobiological studies. One reason is its very robust and predictable rhythms, although the question arises whether this is an inbreeding effect or rather is typical for the species. We compared the daily (circadian) activity rhythms of wild and laboratory golden hamsters. The laboratory hamsters were derived from our own outbred stock (Zoh:GOHA). The wild hamsters included animals captured in Syria and their descendants (F1). Experiments were performed under entrained (light: dark [LD] 14h:10h) and under free-running (constant darkness, DD) conditions. Locomotor activity was recorded using passive infrared detectors. Under entrained conditions, the animals had access to a running wheel for a certain time to induce additional activity. After 3 weeks in constant darkness, a light pulse (15 min, 100 lux) was applied at circadian time 14 (CT14). Both laboratory and wild hamsters showed well-pronounced and very similar activity rhythms. Under entrained conditions, all hamsters manifested about 80% of their total 24h activity during the dark portion of the LD cycle. The robustness of the daily rhythms was also similar. However, interindividual variability was higher in wild hamsters for both measures. All animals used the running wheels almost exclusively during the dark portion of the LD cycle, although the wild hamsters were three times more active. The period length, measured in constant darkness, was significantly shorter in wild (23.93h ± 0.10h) than in laboratory hamsters (24.06 ± 0.07h). The light-induced phase changes were not different (about 1.5h). In summary, these results indicate that the laboratory hamster is not much different from the wild type. (Chronobiology International, 18(6), 921–932, 2001)     

长时间暗适应和弱背景光对鲤鱼视网膜视锥水...

Using intact, immobilized carp preparations, changes in light responsiveness of cone horizontal cells and ultrastructures of their terminals in cone pedicles (HCTs) were correlatively examined in prolonged (greater than 2 h) darkness and after the presentation of a dim background light. Following background illumination cone horizontal cells exhibit high light responsiveness and HCTs give rise to a lot of long, fingerlike or ball-like extensions, called slender or round spinules. When the retina is left in the dark for more than 2 h, the light responsiveness of these cells is depressed, which is accompanied by a dramatic decrease of spinules. Thus light responsiveness of the cone horizontal cells seems to be well correlated with the number of spinules. The results suggest that spinules may play an important role in regulating light responsiveness of cone horizontal cells following background illumination by altering the efficacy of signal transfer across the synapses between cone photoreceptors and cone horizontal cells.     

ACTIVITY RHYTHMS OF WILD AND LABORATORY GOLDEN HAMSTERS (MESOCRICETUS AURATUS) UNDER ENTRAINED AND FREE-RUNNING CONDITIONS

The golden hamster (Mesocricetus auratus) is one of the most frequently used laboratory animals, particularly in chronobiological studies. One reason is its very robust and predictable rhythms, although the question arises whether this is an inbreeding effect or rather is typical for the species. We compared the daily (circadian) activity rhythms of wild and laboratory golden hamsters. The laboratory hamsters were derived from our own outbred stock (Zoh:GOHA). The wild hamsters included animals captured in Syria and their descendants (F1). Experiments were performed under entrained (light: dark [LD] 14h:10h) and under free-running (constant darkness, DD) conditions. Locomotor activity was recorded using passive infrared detectors. Under entrained conditions, the animals had access to a running wheel for a certain time to induce additional activity. After 3 weeks in constant darkness, a light pulse (15 min, 100 lux) was applied at circadian time 14 (CT14). Both laboratory and wild hamsters showed well-pronounced and very similar activity rhythms. Under entrained conditions, all hamsters manifested about 80% of their total 24h activity during the dark portion of the LD cycle. The robustness of the daily rhythms was also similar. However, interindividual variability was higher in wild hamsters for both measures. All animals used the running wheels almost exclusively during the dark portion of the LD cycle, although the wild hamsters were three times more active. The period length, measured in constant darkness, was significantly shorter in wild (23.93h ± 0.10h) than in laboratory hamsters (24.06 ± 0.07h). The light-induced phase changes were not different (about 1.5h). In summary, these results indicate that the laboratory hamster is not much different from the wild type. (Chronobiology International, 18(6), 921-932, 2001)