About the Existence of Circadian Activity in Cave Crayfish

Evidence of a circadian clock mechanism was found in the cave crayfish Procambarus cavernicola. Analysis of motor activity recorded in this species during 12 consecutive days in either free running (constant darkness, DD or constant light, LL) or entrainment conditions (12 h of light alternated with 12 h of darkness, 12 : 12 LD) showed a well recognized circadian rhythm. In this rhythm however, the absence of synchronization by periodical external signals was notorious. The comparison between the motor circadian rhythm in cave crayfish and epigeous crayfish Procambarus clarkii (these last studied during juvenile and adult stages), evidenced strong similitude between the motor circadian rhythm of cave crayfish and juvenile epigeous crayfish.     

Effect of Skeleton Photoperiods upon the Circadian Locomotor Activity Rhythm During Development in Crayfish Procambarus clarkii

This study was carried out to investigate the non-parametric effect of white light on the activity rhythm of the postembryonic stages of crayfish and to elucidate the characteristics and development of its entrainment mechanisms. Forty-five postembryonical instars of crayfish Procambarus clarkii, separated into three groups according to age and moult, were investigated. They were individually monitored with a motor activity recording system in constant photic conditions. For the three groups of animals, three trials with different skeleton photoperiod (SP) imitating LD 8:16, LD 12:12 and LD 20:4 were conducted, each followed by exposure to a period of constant darkness (DD). The locomotor activity rhythm and its entrainment ability were evaluated. All ages showed a circadian rhythm with a capability to synchronize to the different SP increasing with age. The three groups of crayfish analysed were generally synchronized by the dusk pulse, clustering the onset of activity close to the light pulses considered as dusk. The results presented in this study suggest the involvement of extraretinal pathways in the non-parametric entrainment of crayfish.     

Effect of Skeleton Photoperiods upon the Circadian Locomotor Activity Rhythm During Development in Crayfish Procambarus clarkii

This study was carried out to investigate the non-parametric effect of white light on the activity rhythm of the postembryonic stages of crayfish and to elucidate the characteristics and development of its entrainment mechanisms. Forty-five postembryonical instars of crayfish Procambarus clarkii, separated into three groups according to age and moult, were investigated. They were individually monitored with a motor activity recording system in constant photic conditions. For the three groups of animals, three trials with different skeleton photoperiod (SP) imitating LD 8:16, LD 12:12 and LD 20:4 were conducted, each followed by exposure to a period of constant darkness (DD). The locomotor activity rhythm and its entrainment ability were evaluated. All ages showed a circadian rhythm with a capability to synchronize to the different SP increasing with age. The three groups of crayfish analysed were generally synchronized by the dusk pulse, clustering the onset of activity close to the light pulses considered as dusk. The results presented in this study suggest the involvement of extraretinal pathways in the non-parametric entrainment of crayfish.     

Circadian rhythm in melatonin release as a mechanism to reinforce the temporal organization of the circadian system in crayfish

Melatonin (MEL) is a conserved molecule with respect to its synthesis pathway and functions. In crayfish, MEL content in eyestalks (Ey) increases at night under the photoperiod, and this indoleamine synchronizes the circadian rhythm of electroretinogram amplitude, which is expressed by retinas and controlled by the cerebroid ganglion (CG). The aim of this study was to determine whether MEL content in eyestalks and CG or circulating MEL in hemolymph (He) follows a circadian rhythm under a free-running condition; in addition, it was tested whether MEL might directly influence the spontaneous electrical activity of the CG. Crayfish were maintained under constant darkness and temperature, a condition suitable for studying the intrinsic properties of circadian systems. MEL was quantified in samples obtained from He, Ey, and CG by means of an enzyme-linked immunosorbent assay, and the effect of exogenous MEL on CG spontaneous activity was evaluated by electrophysiological recording. Variation of MEL content in He, Ey, and CG followed a circadian rhythm that peaked at the same circadian time (CT). In addition, a single dose of MEL injected into the crayfish at different CTs reduced the level of spontaneous electrical activity in the CG. Results suggest that the circadian increase in MEL content directly affects the CG, reducing its spontaneous electrical activity, and that MEL might act as a periodical signal to reinforce the organization of the circadian system in crayfish.     

Pigment dispersing hormone modulates spontaneous electrical activity of the cerebroid ganglion and synchronizes electroretinogram circadian rhythm in crayfish Procambarus clarkii

In crayfish, one very well-studied circadian rhythm is that of electroretinogram (ERG) amplitude. The cerebroid ganglion has been considered a plausible site for the circadian pacemaker of this rhythm and for the retinular photoreceptors, as the corresponding effectors. The pigment dispersing hormone (PDH) appears to synchronize ERG rhythm, but its characterization as a synchronizer cue remains incomplete. The main purposes of this work were a) to determine whether PDH acts on the cerebroid ganglion, and b) to complete its characterization as a non-photic synchronizer. Here we show that PDH increases the number of the spontaneous potentials of the cerebroid ganglion, reaching 149.92±6.42% of the activity recorded in the controls, and that daily application of PDH for 15 consecutive days adjusts the ERG circadian rhythm period to 24.0±0.2h and the end of the activity period of the rhythm coincides with the injection of the hormone. In this work, we hypothesized that in crayfish, PDH transmits the "day" signal to the ERG circadian system and acts upon both the presumptive circadian pacemaker and the corresponding effectors to reinforce the synchronization of the system.     

Pigment dispersing hormone generates a circadian response to light in the crayfish, Procambarus clarkii

Photoreceptor cells have been identified as important structures in the organization of the circadian system responsible for the generation and expression of the electroretinogram (ERG) circadian rhythm. They are the structures where the circadian periodicity is expressed (effectors) and which transform information from external light signals to be conducted to the pacemaker in order to induce adjustments of the rhythm (synchronizers). After isolation, eyestalks perfused in a pigment dispersing hormone (PDH) solution, show significant changes in receptor potential (RP) amplitude and duration. Exogenous PDH injected into intact crayfish induces a migration of retinal shielding pigments to a light-adapted state. A single dose of PDH produces advances or delays in the circadian rhythm of response to light of visual photoreceptors. All these effects depend on the circadian phase of PDH application. Consequently, the determination of the action of exogenous PDH on photoreceptor cells proved to be very helpful in understanding some mechanisms underlying the circadian organization of crayfish.     

Pigment dispersing hormone modulates spontaneous electrical activity of the cerebroid ganglion and synchronizes electroretinogram circadian rhythm in crayfish Procambarus clarkii

In crayfish, one very well-studied circadian rhythm is that of electroretinogram (ERG) amplitude. The cerebroid ganglion has been considered a plausible site for the circadian pacemaker of this rhythm and for the retinular photoreceptors, as the corresponding effectors. The pigment dispersing hormone (PDH) appears to synchronize ERG rhythm, but its characterization as a synchronizer cue remains incomplete. The main purposes of this work were a) to determine whether PDH acts on the cerebroid ganglion, and b) to complete its characterization as a non-photic synchronizer. Here we show that PDH increases the number of the spontaneous potentials of the cerebroid ganglion, reaching 149.92 ± 6.42% of the activity recorded in the controls, and that daily application of PDH for 15 consecutive days adjusts the ERG circadian rhythm period to 24.0 ± 0.2 h and the end of the activity period of the rhythm coincides with the injection of the hormone. In this work, we hypothesized that in crayfish, PDH transmits the “day” signal to the ERG circadian system and acts upon both the presumptive circadian pacemaker and the corresponding effectors to reinforce the synchronization of the system.     

Shortened seasonal photoperiodic cycles accelerate aging of the diurnal and circadian locomotor activity rhythms in a primate

The gray mouse lemur (Microcebus murinus), a prosimian primate, exhibits seasonal rhythms strictly controlled by photoperiodic variations. Previous studies indicated that longevity can be altered by long-term acceleration of seasonal rhythms, providing a model for assessing various aspects of aging. To assess the effect of aging and accelerated aging on the circadian system of this primate, we compared the circadian rhythm of the locomotor activity in adult mouse lemurs (2-4.5 years, n = 9), aged mouse lemurs (5-9 years, n = 10), and adult mouse lemurs that had been exposed from birth to a shortened seasonal photoperiodic cycle (2-4.5 years, n = 7). Compared to adult animals, aged mouse lemurs showed a significant increase in intradaily variability and an advanced activity onset. Aging was characterized by a decrease in amplitude, with both a decrease in nocturnal activity and an increase in daytime activity. When maintained in constant dim red light, aged animals exhibited a shortening of the free-running period (22.8 +/- 0.1 h) compared to adult animals (23.5 +/- 0.1 h). A 3- to 5-year exposure to an accelerated seasonal photoperiodic rhythm ("annual" duration of 5 months) in accelerated mouse lemurs produced disturbances of the locomotor activity rhythm that resembled those of aged mouse lemurs, whether animals were studied in entrained or in free-running conditions. The present study demonstrated a weakened and fragmented locomotor activity rhythm during normal aging in this primate. Increasing the number of expressed seasonal cycles accelerated aging of parameters related to circadian rhythmicity in adult animals.     

Photoperiodic induction of ovarian maturation in crayfish Procambarus clarkii is mediated by extraretinal photoreception

The current study was carried out to investigate whether the photoperiodic induction of ovarian maturation in crayfish is based on a photosensitive rhythm related to extraretinal photoreceptors. To test this, two batches of 61 juvenile crayfish Procambarus clarkii consisting of [1] intact organisms and [2] animals lacking retina and lamina were exposed to 24h light-dark cycles of different photoperiodic schedules based on a night-break protocol for 3 months. Both batches of crayfish showed the greatest ovarian maturation (size, color, degree and size of oocytes) when the light pulse interrupted the scotophase at 21:00 and 05:00, showing a bimodal photoinducible rhythm. Results of the current study indicate that crayfish ovarian maturation depends on a photoinducible rhythm with two possible states that is related to the circadian clock of crayfish. This phenomenon is mediated by extraretinal photoreceptors. Results are interpreted in the light of models of external coincidence. (Chronobiology International, 18(3), 423-434, 2001)     

Study of circadian activity in the crayfish <Emphasis Type="Italic">Pontastacus Leptodactylus</Emphasis> during their multimonth maintenance in the river water flow

By the method of non-invasive on-line recording and processing of photoplethysmograms of testaceous invertebrates, the circadian rhythm of cardioactivity was studied in crayfish Pontastacus leptodactylus by recording for several month of the heart rate (HR) and stress-index (characteristics of variational pulsometry). The crayfish were kept in the natural running water in regime of the natural illumination alternation (the first group) or at constant artificial illumination of low intensity (the second group). The circadian rhythm was more frequent and more pronounced in crayfish of the first group. The criteria were established to determine the appearance and stabilization of the nocturnal, active rhythm phase: an increase of HR by more than 30% as compared with the daytime rest period and duration of such increase for at least 2.5 h. The stress-index has been shown to be a reliable parameter of the beginning of the nocturnal phase of cardioactivity, while preservation of the typical circadian rhythm can be considered as a bioindicator in the biomonitoring systems of the quality of surface waters.     

Effect of Light Intensity on Circadian Activity in Developing Japanese Quail

The effects of constant light on the expression of the circadian rhythm of feeding activity in Japanese quail, and in particular on the clarity of the rhythm were investigated. We used 46 4-week-old birds (35 females and 11 males) issued from two lines selected for a more (line R: 25 females and 10 males) or less (line A: 10 females and 1 male) clear circadian rhythm of feeding activity. The birds, were placed successively under three light schedules: constant darkness (DD), constant dim green light (LLdim) and constant bright light (LLbright). Schedules were changed every 2 weeks. Feeding activity was recorded continuously, analysed by autocorrelation and spectral analysis, and the ratio of the correlation coefficients and the area of the spectrum peak were used as indexes to quantify the clarity of the circadian rhythm. During the experiment, some birds showed gonadal development. Therefore, we analysed separately birds showing either a high or low degree of sexual development at the end of the experiment. In DD, 35 birds showed a circadian feeding rhythm with a mean period of 22.5 ± 0.1 h, whereas 11 birds showed an arrhythmic activity. In LLdim, 27 birds were rhythmic (22 birds R and 5 birds A), and in LLbright, only 3 birds showed a rhythmic circadian activity. For the R-line birds (for females and males), the rhythm clarity decreased in LLdim compared to DD, except for the not developed females. For the A-line birds (for females), the rhythm clarity of the immature birds increased in LLdim and that of the developed birds remained stable. In LLbright, circadian activity became arrhythmic. In LLdim, the active phases of 12 birds showed two main peaks, with mean periods of 22.7 h and 25.1 h, respectively. Therefore, constant light appeared to have an inhibitory effect on the expression of the circadian rhythm. We postulate that two hierarchically coupled oscillators could control circadian feeding activity, and arrhythmia in LLbright could be the results of internal desynchronization of the pacemakers.     

Effect of Light Intensity on Circadian Activity in Developing Japanese Quail

The effects of constant light on the expression of the circadian rhythm of feeding activity in Japanese quail, and in particular on the clarity of the rhythm were investigated. We used 46 4-week-old birds (35 females and 11 males) issued from two lines selected for a more (line R: 25 females and 10 males) or less (line A: 10 females and 1 male) clear circadian rhythm of feeding activity. The birds, were placed successively under three light schedules: constant darkness (DD), constant dim green light (LLdim) and constant bright light (LLbright). Schedules were changed every 2 weeks. Feeding activity was recorded continuously, analysed by autocorrelation and spectral analysis, and the ratio of the correlation coefficients and the area of the spectrum peak were used as indexes to quantify the clarity of the circadian rhythm. During the experiment, some birds showed gonadal development. Therefore, we analysed separately birds showing either a high or low degree of sexual development at the end of the experiment. In DD, 35 birds showed a circadian feeding rhythm with a mean period of 22.5 ± 0.1 h, whereas 11 birds showed an arrhythmic activity. In LLdim, 27 birds were rhythmic (22 birds R and 5 birds A), and in LLbright, only 3 birds showed a rhythmic circadian activity. For the R-line birds (for females and males), the rhythm clarity decreased in LLdim compared to DD, except for the not developed females. For the A-line birds (for females), the rhythm clarity of the immature birds increased in LLdim and that of the developed birds remained stable. In LLbright, circadian activity became arrhythmic. In LLdim, the active phases of 12 birds showed two main peaks, with mean periods of 22.7 h and 25.1 h, respectively. Therefore, constant light appeared to have an inhibitory effect on the expression of the circadian rhythm. We postulate that two hierarchically coupled oscillators could control circadian feeding activity, and arrhythmia in LLbright could be the results of internal desynchronization of the pacemakers.     

Residual circadian rhythmicity after bilateral lamina-medulla removal or optic stalk transection in the cricket, Gryllus bimaculatus

Bilateral optic stalk severance or lamina-medulla region removal were carried out in 47 adult male crickets Gryllus bimaculatus DeGeer. Effects of the operations on circadian locomotor activity were investigated under 12 h light: 12 h dark and at a constant temperature of 26°C. In the pre-operative days, 39 of the animals showed a typical nocturnal activity rhythm (normal rhythm), but the remaining 8 exhibited an atypical rhythm which is diurnal rather than nocturnal (abnormal rhythm). The operations eventually caused an arrhythmicity in all animals, suggesting that the crucial part of the central nervous system controlling the cricket circadian activity is located in the lamina-medulla region. However, in some of the post-operative crickets, the rhythm did not immediately disappear but persisted for a while: the diurnal increase of activity was observed up to 2 weeks in all 8 abnormal- and 4 normal-rhythm animals. In addition, 8 out of 39 normal-rhythm animals showed a single well-defined post-operative peak which occurred approximately in phase with the nocturnal peak prior to surgery. These results are discussed in relation to a possibility of involvement of the oscillatory structure outside the optic lobes.     

Erg circadian rhythm in the course of ontogeny in crayfish

• 1.1. The objective of the present work was to study the ontogeny of the ERG circadian rhythm in crayfish.
• 2.2. Long-term recordings of ERG and shielding retinal pigments position measured from the instar, the second instar, the third instar and the adult crayfish were obtained.
• 3.3. In the youngest animals (1–8 days old) an ultradian rhythm (15min-4hr periods) in the ERG amplitude was detected.
• 4.4. Older animals showed a progressive increment in the period length before they exhibited a circadian pattern. This last appeared, the first time, in 30-day-old animals and showed noticeable differences in the adult crayfish. At the same time, the crayfish began to show photomotor reflex. Later on (140-day-old crayfish) the circadian rhythm attained its final parameters.
• 5.5. The SD was used as a measure of lability in periods. The 4 hr ultradian rhythm and the 22.4 hr circadian rhythm showed the lowest SD indicating that they are the most precise period values.
• 6.6. Our results support the idea that the ERG circadian rhythm results from the coupling among high frequency (ultradian) oscillators, particularly those of 4 hr periods and that the coupling depends on the action of neurosecretions released from the sinus gland.

     

Circadian rhythms of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), insulin-like growth factor-1 (IGF-1), insulin-like growth factor binding protein-3 (IGFBP-3), cortisol, and melatonin in women with breast cancer

Background: Circadian rhythms in plasma concentrations of many hormones and cytokines determine their effects on target cells. Methods: Circadian variations were studied in cortisol, melatonin, cytokines (basic fibroblast growth factor [bFGF], EGF, insulin-like growth factor-1 [IGF-1]), and a cytokine receptor (insulin-like growth factor binding protein-3 [IGFBP-3]) in the plasma of 28 patients with metastatic breast cancer. All patients followed a diurnal activity pattern. Blood was drawn at 3h intervals during waking hours and once during the night, at 03:00. The plasma levels obtained by enzyme-linked immunoassay (ELISA) or radioimmunoassay (RIA) were evaluated by population mean cosinor (using local midnight as the phase reference and by one-way analysis of variance (ANOVA). Results: Cortisol and melatonin showed a high-amplitude circadian rhythm and a superimposed 12h frequency. bFGF showed a circadian rhythm with an acrophase around 13:00 with a peak-to-trough interval (double amplitude) of 18.2% and a superimposed 12h frequency. EGF showed a circadian rhythm with an acrophase around 14:20, a peak-to-trough interval of 25.8%, and a superimposed 12h frequency. IGF-1 showed a high value in the morning, which is statistically different t test) from the low value at 10:00, but a regular circadian or ultradian rhythm was not recognizable as a group phenomenon. IGFBP-3 showed a low-amplitude (peak-to-trough difference 8.4%) circadian rhythm with the acrophase around 11:00 and low values during the night. Conclusions: (1) Circadian periodicity is maintained in hospitalized patients with metastatic breast cancer. (2) Ultradian (12h) variations were superimposed on the circadian rhythms of the hormones and several of the cytokines measured. (3) Studies of hormones and cytokines in cancer patients have to take their biologic rhythms into consideration. (4) The circadian periodicity of tumor growth stimulating or restraining factors raises questions about circadian and/ or ultradian variations in the pathophysiology of breast cancer. (Chronobiology International, 18(4), 709-727)     

In situ studies on heart rate and locomotor activity in the freshwater crayfish, Astacus astacus (L.) in relation to natural fluctuations in temperature and light intensity

1. By the aid of a computer technique, the in situ variations in heart rate and locomotor activity were recorded continuously for 7 days in sixteen individuals of the crayfish, Astacus astacus (Linnaeus 1758), in association with natural changes in water temperature and light intensity.
2. All sixteen crayfish investigated were found to express circadian rhythmicity in heart rate with a periodicity of ≈ 24 h. For 88% of the animals investigated, rhythmicity was significant ( P < 0.05). Ninety‐three per cent of the crayfish showed circadian rhythmicity in locomotor activity with a 24 ± 2 h periodicity and, for 67% of the animals, this rhythmicity was significant ( P < 0.05). In general, the expression of circadian rhythms was associated with nocturnal behaviour, heart rates and locomotor activity levels being higher at night than by day.
3. A positive correlation was observed between heart rate and locomotor activity. Temperature and light intensity exerted positive and negative influences on heart rate in A. astacus , respectively.
4. In the first experiment, 84% of the variation in heart rate could be explained by the changes in locomotor activity, temperature and light intensity. In the second experiment 35% of the variation in the heart rate was explained using these parameters.     

Daily and circadian rhythms in the activity of mixed function oxidases system in rats of different age

Abstract

An age‐associated alterations in daily and circadian changes of cytochrome P‐450‐linked monooxygenases system activity were studied using 1‐, 6‐ and 12‐months old male Wistar rats, in winter and in spring season. Cytochrome P‐450 and NADPH‐cytochrome P‐450 reductase showed 12h rhythm in all investigated age groups of animals. Cytochrome b₅ and NADH‐cytochrome b₅ reductase were characterized by a 12h daily rhythm in 1‐month old rats, but in older ones 24h circadian rhythm was found. There was not significant changes of the rhythm pattern in the activity of investigated MFO system ingredients in rats of different age, between spring and winter.     

Cerebroid Ganglion is the Presumptive Pacemaker of the Circadian Rhythm of Electrical Response to Light in the Crayfish

One of the most widely studied circadian rhythms in invertebrates is that of light responsiveness whose underlying mechanisms seem to involve different groups of oscillators which act as pacemakers. Although, in crayfish, there are clear circadian rhythms in the electroretinogram (ERG) amplitude, the precise location of the pacemaker system driving this rhythm is uncertain. Some data suggest that the circadian pacemaker could be located in a group of neurosecretory cells of the supraesophageal ganglion (the cerebroid ganglion or brain) and that the sinus gland plays a determinant role in the generation and expression of this rhythm through periodic release of pigment-dispersing hormone (PDH). The aim of this work is to examine the role of the brain in the expression of the ERG circadian activity. The hypothesis we test is that the electrical activity at the brain level has a circadian behavior in the firing pattern of spontaneous multiunit activity (MUA) and in visual evoked potentials (VEPs). The results indicate that there are robust circadian rhythms in both MUA, recorded from several regions of the brain, and in the averaged VEPs recorded from the protocerebrum area. These rhythms are 180° out of phase to one another. The rhythm of VEPs showed a main peak at midnight which was in close phase relationship with the ERG amplitude rhythm.     

Spontaneous and light-evoked discharge of the isolated abdominal nerve cord of crayfish in vitro reveals circadian oscillations.

We examined the well-known spontaneous discharge (SD) and light-evoked (PD) discharge of the crayfish caudal photoreceptor for the possible existence of a daily rhythm in spike frequency. To do this, we isolated the abdominal nerve cord in vitro and studied its discharge frequency in constant darkness. Single cosinor analysis revealed significant SD and PD circadian rhythms (P < .05) with periods tau = 24.4h and 24.2h, respectively. These oscillations correspond to an endogenous circadian discharge of the caudal photoreceptor that is enhanced by light. The importance of this rhythm in the adaptive behavior of crayfish is discussed.     

Circadian and seasonal responses in Indian weaver bird: subjective interpretation of day and night depends upon both light intensity and contrast between illuminations

This study investigated whether changes in illumination modify perception of day and night conditions in a diurnal species, the Indian weaver bird. Birds were initially subjected to a 12-h light:12-h dark regime (12L:12D; L=20 lux, D =0.5 lux). After every 2 wks, the combinations of light illumination in L and D phases were changed as follows: 20:2 lux, 20:5 lux, 20:10 lux, 20:20 lux, 20:100 lux, and 20:200 lux. Finally, birds were released into dim constant light (0.5 lux) for 2 wks to determine the phase and period of the circadian activity rhythm. They were also laparotomized at periodic intervals to examine the effects of the light regimes on the seasonal testicular cycle. All individuals showed a consistently similar response. As evident by the activity pattern under these light regimes, both in total activity during contrasting light phases and during the 2?h in the beginning and end of first light phase, birds interpreted the period of higher light intensity as day, and the period of lower intensity as the night. During the period of similar light intensity, i.e., under LL, birds free-ran with a circadian period ( ~ 24 h). In bright LL (20 lux), the activity rhythm was less distinct, but periodogram analysis revealed the circadian period for the group as 24.46 (+/-) 0.41 h (mean???SE). However, in dim LL at the end of the experiment, all birds exhibited a circadian pattern with average period of 25.52 (+/-) 0.70 h. All birds also showed testicular growth and regression during the 16-wks study. It is suggested that weaver birds interpret day and night subjectively based on both the light intensity and contrast between illuminations during two phases over the 24 h.