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.     

Melatonin modulates the ERG circadian rhythm in crayfish

One of the most important functions modulated by melatonin is the synchronization of circadian rhythms. In crayfish (Procambarus clarkii), we have obtained evidence that the amplitude of the electrical response to light of the retinal photoreceptors the receptor potential, is modified by the action of melatonin and that the magnitude of this action depends on the circadian time of melatonin application. In contrast, the electroretinogram (ERG) circadian rhythm can be synchronized by either single or periodic melatonin application. In this work we hypothesized that, in crayfish, melatonin acts on effectors and on pacemaker of ERG circadian rhythm as a non-photic synchronizer. Melatonin could be a hormone that sends a signal of darkness to the ERG circadian system.     

Reciprocal neural influences upon the period length of the ERG circadian rhythm in the crayfish

Abstract

The relationship between left and right circadian oscillations in the ERG of the crayfishProcambarus bouvieri have been studied in intact animals and in animals with cerebral ganglion damage. Except in split‐brain preparations, the ERG oscillations of both sides were always in phase. Surgical bisection of the cerebral ganglion, also results in a shortening of the period length; this suggests that ERG circadian pacemakers normally receive a bilateral input of light stimulation. The neural connections between the eyestalks at the protocerebrum seem to have the most important role in the synchronization of the ERG oscillations of both sides.     

Synchronization by light of the circadian rhythm of motor activity in the crayfish

Abstract

We have studied the pattern for resetting the circadian rhythm in the spontaneous motor activity of the crayfish. Spontaneous motor activity was recorded continously at a constant temperature and under free running conditions in complete darkness. The effect of single light pulses applied at different circadian times, on the circadian rhythm of motor activity was measured in both transient stage and steady state. The results led us to construct a phase‐transition curve and phase‐response curve which were analyzed to obtain information about the oscillators which underlie the circadian rhythm of motor activity.     

Expression of genes encoding Ca exporting proteins in freshwater crayfish Procambarus clarkii during cold exposure

1.

This study examined expression of two primary transmembrane Ca²⁺ export proteins (plasma membrane Ca²⁺ ATPase, (PMCA); Na⁺/Ca²⁺ exchanger, sodium/calcium exchanger (NCX)) in epithelial (antennal gland, kidney) and non-epithelial (axial abdominal muscle) tissues of the freshwater crayfish Procambarus clarkii following exposure (28 days) to 4 °C (compared with control 23 °C).     

Circadian clock and prothoracicotropic hormone secretion in relation to the larval-larval ecdysis rhythm of the saturniid Samia cynthia ricini

Observations on the timing of ecdysis and neck ligation experiments on larvae of Samia cynthia ricini under various light-dark conditions show that an endogenous circadian clock controls the timing of larval ecdysis and prothoracicotropic hormone (PTTH) secretion preceding it. The clock, upon reaching a specific phase point, causes the brain to secrete PTTH provided that the brain has acquired the secretory competence. This time may vary, in relation to a previous ecdysis, according to the light-dark conditions by which the clock phase is specifically determined, but is fixed relative to a subsequent ecdysis. Thus, in the case of the ecdysis to the 5th instar, PTTH is secreted [15+nτ] hr (τ: free-running period, slightly less than 24 hr) after the clock has started when the rhythm is free-running, and in the second and third nights of the 4th instar under a photoperiod of 12 hr light and 12 hr dark. Full secretion of ecdysone occurs 6 hr after PTTH secretion and ecdysis ensues 34 hr thereafter to complete the ultimate sequence of ecdysis.     

Development of the circadian clock in the German cockroach, Blattella germanica

The cell distribution and immunoreactivity (ir) against period (PER), pigment dispersing factor (PDF) and corazonin (CRZ), were compared between adults and nymphs in the central nervous system of the German cockroach. Although PER-ir cells in the optic lobes (OL) were expressed in the nymphs from the first instar, the links between major clock cells became more elaborated after second/third instar. A circadian rhythm of locomotion was initiated at the fourth/fifth instar. The results suggest that the clock was running from hatching, but the control network needed more time to develop. In addition, the putative downstream regulators, PDF-ir and CRZ-ir, are co-localized in various regions of the brain, indicating potential output routes of the circadian clock. CRZ-ir cells with typical morphology of neurosecretory cells in the dorsolateral protocerebrum send out three neural fibers to reach the ipsilateral corpora cardiaca (CC), the antennal lobe and two hemispheres of the protocerebrum. Based on co-localization with some PER-ir/PDF-ir cells, the CRZ-ir cells have the potential to serve as a bridge between circadian neural signals and endocrine regulation. Based on PDF's role in the regulation of locomotion, our results support the finding that the locomotor circadian rhythm is possibly controlled by a hormonal route.     

Cyclic melatonin synchronizes the circadian rhythm of feeding activity in Japanese quail,Coturnix c. japonica

In Japanese quail, we can observe the circadian rhythm of feeding activity in constant conditions, especially in birds from selected lines. In order to try to test the importance of melatonin as hormonal output for the circadian system, we gave a 24-h period cycle of exogenous melatonin to some of these birds when they were free running. We used castrated males firstly in order to cancel the known effect of steroids on circadian organisation. Secondly, as castrated birds generally expressed a very short periodicity, it allowed us to check induced synchronisation more easily. We maintained ten castrated males in constant dim light. We divided the experiment into five successive phases. The birds received a 24-h period cycle of melatonin (M phase) or of control solution with only the alcoholic solvent (C phase) as a drink. Before and after each one of these two phases, we gave water continually to drink (W1, W2 and W3 phases). Thus, the successive phases were W1-M-W2-C-W3. We measured intake of liquids and plasma melatonin concentrations to check melatonin ingestion. We automatically recorded individual feeding activity by infrared detectors, and analysed this by spectral analysis. At the beginning of the experiment, eight birds showed a rhythmic feeding activity, with a mean period of 22.9 +/- 0.2 h, and the two others an arrhythmic circadian activity. During the 24-h period cycle of exogenous melatonin, for the rhythmic birds, the circadian period became approximately 24 h (23.9 +/- 0.2 h), the inactive phase corresponding to the period of melatonin availability. During the W2 and C phases, the circadian period was similar to that expressed during the W1 phase. Moreover, when birds only drink water, we found a significant positive relationship between the clarity of the circadian rhythm and the ratio, between the melatonin level of the inactive phase and that of the active phase. These facts support the hypothesis of the role of this hormone in the regulation of the circadian system, at least for feeding activity, in quail.     

Progressive retinal ganglion cell loss in primary open-angle glaucoma is associated with temperature circadian rhythm phase delay and compromised sleep

Advanced primary open-angle glaucoma (POAG) is characterized by progressive retinal ganglion cell complex (RGCC) damage that may cause subsequent disruption of the circadian rhythms. Therefore, we evaluated circadian body temperature (BT) rhythm and sleep characteristics of 115 individuals (38 men and 77 women) diagnosed with POAG. GLV (global loss volume; %), a measure of RGCC damage, was estimated by high-definition optical coherence tomography, and RGC functional ability was assessed by pattern electroretinogram amplitude (PERGA). Depending on dynamics of POAG progression criteria, two groups were formed that were distinctively different in GLV: Stable POAG group (S-POAG; GLV = 5.95 ± 1.84, n = 65) and Progressive POAG group (P-POAG; GLV = 24.27 ± 5.09, n = 50). S-POAG and P-POAG groups were not different in mean age (67.61 ± 7.56 versus 69.98 ± 8.15) or body mass index (24.66 ± 3.03 versus 24.77 ± 2.90). All subjects performed 21 around-the-clock BT self-measurements during a 72-h period and kept activity/sleep diaries. Results showed pronounced disruption of circadian physiology in POAG and its progression with increasing severity of the disease. The daily mean of BT was unusually low, compared to age-matched controls. Moreover, our results revealed distinctive features of BT circadian rhythm alterations in POAG development and POAG progression. S-POAG is associated with lowered BT circadian rhythm robustness and inter-daily phase stability compared to controls. In the P-POAG group, the mean phase of the circadian BT rhythm was delayed by about 5 h and phases were highly scattered among individual patients, which led to reduced group mean amplitude. Circadian amplitudes of individuals were not different between the groups. Altogether, these results suggest that the body clock still works in POAG patients, but its entrainment to the 24-h environment is compromised. Probably because of the internal desynchronization, bedtime is delayed, and sleep duration is accordingly shortened by about 55 min in P-POAG compared to S-POAG patients. In the entire POAG cohort (both groups), later sleep phase and shorter mean sleep duration correlate with the delayed BT phase (r = 0.215; p = 0.021 and r = 0.322; p = 0.0004, respectively). An RGCC GLV of 15% apparently constitutes a threshold above which a delay of the circadian BT rhythm and a shortening of sleep duration occur.     

Effectiveness of dam construction to contain the invasive crayfish Procambarus clarkii in a Mediterranean mountain stream

Invasive crayfish are a worldwide problem for aquatic ecosystems. So far, no control method has proven successful. Field data from invaded streams have suggested that physical structures may hamper the spread rate of invasive crayfish into new areas. However, given the difficulties for carrying out dam construction in natural habitats, data about their effectiveness in incipient invading populations are practically nonexistent. This paper evaluates the effectiveness of small dams to contain the advance of an incipient population of the invasive Procambarus clarkii in a mountain stream harboring the European southernmost population of the endangered Austropotamobius pallipes. The management strategy involved the construction of 3 artificial small (<3 m high) dams that were complemented with specific design solutions to increase their theoretical efficacy. Catch Per Unit Effort (CPUE) obtained during monitoring surveys carried out for 4 years were used as an indicator of containment effectiveness. The dams successfully prevented Procambarus clarkii from extending its range into the area occupied by A. pallipes. In addition, regardless of whether the construction of new dams isolated the leading edge of the invading population between two dams, the CPUE declined to below detectable limits. Therefore, dams are suggested to effectively stop, or at least hinder, the advance of invasive crayfish. Long-term monitoring is recommended to verify the on-going success of the dams in preventing upstream invasion.     

Eclosion hormone activity in developing embryos of the silkworm, Bombyx mori

The ultimate timing of hatching in the silkworm, Bombyx mori, is controlled by a circadian oscillator. The presence of eclosion hormone in developing embryos of the silkworm is demonstrated. Eclosion hormone activity first becomes detectable in embryos which have developed almost to the stage of the differentiation of the neuroendocrine system. Hormonal activity increases sharply to a maximum level 1 day before hatching and falls by about a half in the newly hatched larvae. Eclosion hormone was partially purified from the pharate first-instar larvae and approx, a 2100-fold purification was achieved. The molecular weight of the embryo eclosion hormone is estimated to be 7000 ～ 9000 Daltons by gel-filtration on Sephadex G-50 (superfine). The role of eclosion hormone on hatching behaviour of the silkworm, Bombyx mori, is discussed.     

Characteristics of the circadian activity rhythm in common marmosets (Callithrix j. jacchus)

The circadian activity rhythm of the common marmoset, Callithrix j. jacchus was investigated by long-term recording of the locomotor activity of 15 individuals (5 males, 10 females) from 1.5 to 8 years old, both under constant illumination and under LD 12:12. The mean period of the spontaneous circadian rhythm was 23.2 ± 0.3 h. Neither sex-specific differences nor a systematic influence of light intensity on the spontaneous period were observed, but the period was dependent on the duration of the trial and on the age of the individual. Due to the short spontaneous period, in LD 12:12 there was a distinct advance of the activity phase with respect to the light time and a masking of the true onset of activity by the inhibitory direct effect of low light intensity during the dark time. After an 8 h delay shift of the LD 12:12, re-entrainment of the circadian activity rhythm required an average of 6.8 ± 0.7 days; the average re-entrainment time after an 8 h phase advance of the LD cycle was 8.6 ± 1.3 day. This directional effect is ascribed to characteristics of the phase-response curve. No ultradian components were observed, either in the LD-entrained or the free-running circadian activity rhythm.     

Lack of a circadian rhythm in the ability of the gametocytes of Plasmodium falciparum to infect Anopheles gambiae

Batches of Anopheles gambiae were fed once during the day or once or twice during the night on 12 carriers naturally infected with gametocytes of P. falciparum. No overall difference was noted in the oocyst numbers in batches of mosquitoes fed during the night as compared to the daytime.     

AFLP-PCR and RAPD-PCR evidences of the transmission of the pathogen Aphanomyces astaci (Oomycetes) to wild populations of European crayfish from the invasive crayfish species, Procambarus clarkii

Aphanomyces astaci (Oomycetes) is responsible for the crayfish plague disease. This species is endemic of North America and five genotypes have been described using RAPD-PCR. The red swamp crayfish, Procambarus clarkii, is one of the most widely spread North American species and invasive in the world. However, no outbreaks on its specific genotype, i.e., genotype D, have ever been described in nature. We investigated three major series of crayfish plague outbreaks in indigenous crayfish populations of Austropotamobius pallipes, located in the areas of influence of P. clarkii. All samples collected tested positive for A. astaci using a rnDNA ITS-PCR test. We also performed an AFLP-PCR analysis on 19 isolates, and found that all isolates belong to genotype D. These isolates exhibited similar properties, i.e., adaptation to warm temperatures. We demonstrate, for the first time, the transmission of A. astaci genotype D to indigenous European populations of crayfish, and confirm that the properties of adaptation to warm water temperatures seem to be a specific character of genotype D. The results of this work emphasize once more the need of controlling invasive species and its trade, since they can carry harmful pathogens with specific adaptations or increased virulence in new environments.     

Effects of weight, temperature and behaviour on the circadian rhythm of salivary cortisol in growing pigs

In farm animals, salivary cortisol has become a widely used parameter for measuring stress responses. However, only few studies have dealt with basal levels of concentration of cortisol in pigs and its circadian rhythm. The aim of this study was to examine the effects of ambient temperature and thermoregulatory behaviour on the circadian rhythm of salivary cortisol levels in fattening pigs. Subjects were 30 fattening pigs of different weight (60 to 100 kg), kept in six groups in an uninsulated building in pens with partly slatted floors. Saliva samples were taken every 2 h over periods of 24 h at different ambient temperatures at two times in winter and four times in summer. Thermoregulatory behaviour was recorded in the same 24-h time periods. The effect of time of day, body weight, ambient temperature and behaviour on the cortisol level was analysed using a mixed-effects model. Two peaks of cortisol levels per day were found. This circadian pattern became more pronounced with increasing weight and on days where thermoregulatory behaviour was shown. Mean cortisol levels per day were affected by weight but not by thermoregulatory behaviour. From our data, we conclude that long-term variations in cortisol concentration may be influenced by increasing age and weight more than by the respective experimental situation. In assessing animal welfare, it seems more reliable to consider the circadian pattern of cortisol concentration instead of only one value per day.     

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.     

Temporal organization of endocrine events in relation to the circadian clock during larval-pupal development in Samia cynthia ricini

The temporal organization of secretion of the prothoracicotropic hormone (PTTH) and ecdysone during larval-pupal development of Samia cynthia ricini was studied by ligations, with particular attention to the circadian control of the timing of hormone release. PTTH and ecdysone are required first for the induction of prodromes of pupation and again later for pupal-cuticle formation. PTTH release in the first step occurs during the second or third photophase after the last-larval ecdysis under a photoperiod of 12 hr light and 12 hr darkness and is thought to be under the control of a circadian clock. Ecdysone release follows 1.5 days later, i.e. during the scotophase that precedes the gut purge. In the second secretory step, PTTH is released 2 days after purging the gut, and ecdysone release follows 6 hr later. The PTTH release at this time occurs at a fixed time after the gut purge irrespective of light conditions, accounting for light insensitivity of the timing for pupal ecdysis. Possible mechanisms relating to the inconsistent association of a circadian clock with PTTH release, and those underlying the determination of timing of the gut purge are discussed.