Amphetamine-induced locomotor behavior of mice is influenced by DSIP

The delta sleep-inducing peptide (DSIP) has been shown to induce effects other than only delta sleep. One of these effects was the paradoxical thermoregulatory and locomotor response of rats to amphetamine after DSIP administration. In the present investigation we found similar effects of DSIP on the locomotor activity in mice. However, two different doses of DSIP (30 and 120 nmol/kg) and 3 doses of amphetamine (4, 10, and 15 mg/kg) produced a complex pattern of effects in mice tested at 22 degrees C. In general, DSIP-treated mice showed lower locomotor activity after amphetamine than controls, but under two conditions, both using 15 mg/kg amphetamine, DSIP produced higher scores; this occurred in the first two hours after amphetamine for the 30 nmol/kg DSIP group and in the third hour for mice given 120 nmol/kg DSIP. The results indicate that the effects of DSIP on locomotor behavior were dependent on the dosage of the peptide and the time of measurement as well as the level of amphetamine stimulation.     

The delta sleep-inducing peptide as a factor enhancing the content of substance P in the hypothalamus and the resistance of rats to emotional stress]

The influence of the delta-sleep inducing peptide (DSIP, 60 and 120 nmol/kg, intraperitoneally) on the content of substance P (SP) in rats hypothalamus was studied on males of August line. DSIP administration significantly increased the mean SP content in the hypothalamus and also its content in animals, stable and predisposed to emotional stress. Daily DSIP administration before putting the rats in conditions of stress increased the SP content in the hypothalamus decreased at the emotional stress. Preliminary single DSIP administration to the animals subjected to stress also increased the SP content. Single DSIP administration in a dose of 60 nmol/kg sharply reduced classical stress manifestations, such as hypertrophy of adrenals and thymus involution.     

The effect of delta sleep-inducing peptide on the convulsive activity in corasol kindling

The influence of intraperitoneal delta-sleep inducing peptide (DSIP) injection (100 micrograms/kg) on the epileptic activity was investigated in the experiments on Wistar rats and (CBA X C57B1/6)F1 mice. The model of chronically developing epileptic activity--the model of pharmacological kindling--was created by daily repeated corasole injections in subconvulsive doses (30 mg/kg). It has been shown that DSIP injection delayed the manifestation of generalized seizures during kindling, led to the suppression of seizure activity and reduced the mortality rate of animals that developed kindled seizures. The antiepileptic effect of DSIP was observed throughout the period of 5 minutes to 24 hours after the injection. Naloxone (2.5 mg/kg) did not change the antiepileptic effect of DSIP.     

Circadian rhythm of cardiac bradyarrhythmia episodes in rats

It was investigated whether incidence of S-A block and A-V block (type I and II) observed in male Wistar rats shows a circadian periodicity. Under the 14/10 light-dark illumination schedule, circadian periodicity was demonstrated in the occurrence of bradyarrhythmias as well as slow wave sleep, paradoxical sleep locomotive activity and heart beats/hour. This circadian variation of bradyarrhythmia shows two major peaks. The highest peak coincides with the time zone immediately after the start of illumination, and the second peak coincides with the acrophase of the circadian variation in paradoxical sleep. The 25-h period of circadian rhythm of bradyarrhythmias was disclosed even under constant illumination. This circadian variation shows the disappearance of the early peak of two peaks observed in the light-dark schedule. These results show the existence of an endogenous circadian rhythm in bradyarrhythmias as well as in sleep-wakefulness and locomotive activity. This knowledge about the circadian rhythm of bradyarrhythmia must be incorporated into the effective treatment of arrhythmias.     

Locomotor Rhythms in the Fiddler Crab,Uca subcylindrica

The locomotor activities of individual specimens of Uca subcylindrica (Stimpson) collected from semi-arid, supratidal habitats in south Texas and northeastern Mexico were studied in the laboratory using periodogram analysis. When crabs were placed under constant darkness (DD) or constant illumination (LL), free-running circadian rhythms were observed in the activity recordings. The locomotor activity of strongly rhythmic crabs in LL has an average period length of 24.4 h. Crabs held in DD express motor rhythms with periods of approximately 24.0 h. In LL the most common wave form for activity is unimodal, while under DD it is bimodal. Recordings under natural illumination (NL) revealed that both period length and the time of maximum activity (phasing) varied through the year. During winter months, the crabs are primarily diurnal with peaks in activity occurring between 0900 and 2100 h and possess a circadian rhythm with a 23.9 h period. During summer, crabs were nocturnal with maximal activity between 1300 and 0600 and a circadian period closer to 24.0 h. In these experiments, the rhythmic locomotor activities of U. subcylindrica are best described as “circadian”. This is unusual for a genus known for its expression of circatidal and circalunidian rhythms.     

Effects of Illumination on the Circadian Motor Rhythm of the Chelipeds of Crayfish

The effects of different illumination conditions on the main parameters of the circadian motor rhythms of the two chelipeds of the crayfish, Procambarus digueti , were compared. Under either constant darkness (DD) or constant light (LL) the phase relationship between the two circadian rhythms was more stable than under entrained conditions (LD cycles). These results suggest that the oscillators responsible for these rhythms differ in their sensitivity to light. The role of paired organs in the internal temporal order of the crayfish is discussed.     

Effects of Illumination on the Circadian Motor Rhythm of the Chelipeds of Crayfish

The effects of different illumination conditions on the main parameters of the circadian motor rhythms of the two chelipeds of the crayfish, Procambarus digueti, were compared. Under either constant darkness (DD) or constant light (LL) the phase relationship between the two circadian rhythms was more stable than under entrained conditions (LD cycles). These results suggest that the oscillators responsible for these rhythms differ in their sensitivity to light. The role of paired organs in the internal temporal order of the crayfish is discussed.     

Appearance of parkinsonian syndrome after administration of delta sleep-inducing peptide into the rat substantia nigra

Bilateral microinjection of delta-sleep-inducing peptide (DSIP) (10.0 nmol) into the substantia nigra provoked hypokinesia and rigidity in rats observed during 4.0 hours. Injection of DSIP in dose of 5.0 nmol into the substantia nigra or into the nuclei caudati in dose of 10.0 nmol did not induce such symptoms. The enhanced slow-wave activity was recorded in caudate nuclei during hypokinesia and rigidity which demonstrated the formation of the generator of pathologically enhanced excitation (GPEE). The systemically cyclodol administration resulted in abolishment of rigidity and increase in locomotor activity. The conclusion is that bilateral intranigral DSIP injection caused acute parkinson syndrome in rats due to the formation of cholinergic GPEE in caudate nuclei. The hyperactive caudate nuclei act as the pathologic determinant which induces the parkinson syndrome.     

Action of the "delta-sleep peptide" on monoamine oxidase and acetylcholinesterase activity in subcellular fractions from different rabbit brain formations in vivo

Administration of delta-sleep-inducing peptide (DSIP) in vivo in a dose of 30 microgram/kg bw brings about MAO-A (substrate-serotonin) activation in synaptosome subfractions and cellular mitochondria from the brain structures (motor cortex, nucleus caudatus, thalamus). Activity of MAO-B (substrate-p-nitrophenylethylamine) and acetylcholinesterase was inhibited negligibly and specifically in subcellular fractions of the test brain structures. The results suggest that DSIP effects the regulatory or modulation function in the synapse. As one of the elements of sleep mechanisms this peptide induces a number of processes, particularly in serotonin metabolism.     

DSIP affects adrenergic stimulation of rat pineal N-acetyltransferase in vivo and in vitro

The natural occurrence, sleep, and extra-sleep effects of delta sleep-inducing peptide (DSIP) have been shown by different laboratories. However, neither an in vitro assay system nor a probable mechanism of action of the peptide have been conclusively demonstrated so far. The recent finding that DSIP influences the nocturnal rise of N-acetyltransferase (NAT) activity in rat pineal led us to investigate a possible effect on pharmacologically induced NAT activity in vivo and in vitro. Stimulation of the enzyme with adrenergic drugs such as isoproterenol and phenylephrine was reduced by DSIP at doses of 150 and 300 μg/kg injected subcutaneously. In vitro, 6, 150 and 300 nM DSIP attenuated isoproterenol stimulation of the enzyme in cultured pineals, whereas 150 nM DSIP effectively reduced stimulation induced by a combination of the two drugs. The peptide alone did not influence NAT activity in vitro, but produced a slight stimulation in vivo. To our knowledge, these results represent the first report of a direct interaction of DSIP with adrenergic transmission. The in vitro system could prove useful for establishing possible mechanism(s) of action of the ‘sleep peptide.’     

Phosphorylation of Delta Sleep-Inducing Peptide (DSIP) by casein kinase II in vitro

A phosphorylated analogue of DSIP at Ser⁷ has been shown to exist endogenously by immunochemical studies. An enzyme which could phosphorylate DSIP has not yet been identified. In the present study, we examined DSIP as a substrate for in vitro phosphorylation by casein kinase II. DSIP was phosphorylated by the enzyme with apparent K_m and V_max values of 20 mM and 90.9 nmol/min/mg protein, respectively. Both ATP and GTP were utilized as phosphoryl donors. Phosphorylation of DSIP was inhibited by heparin and enhanced by spermine. These results demonstrate that DSIP can serve as a possible substrate for casein kinase II in vitro.     

Pinealectomy and LL Abolished Circadian Perching Rhythms but Did Not Alter Circannual Reproductive or Fattening Rhythms in Finches

In the subtropical finch, spotted munia (Lonchura punctulata) circannual rhythms (of gonads, fattening, feeding) have been demonstrated in an information-free environment of continuous illumination (LL), rendering it an ideal model for research on the physiology of the circannual clock. In an attempt to understand the involvement, if any, of the circadian system in the genesis of circannual rhythms, we studied the effect of pinealectomy (LL 15 lux) and strong continuous illumination (LL 300 lux), both known to abolish circadian rhythms, on the circadian perch-hopping rhythm and on the circannual rhythm of reproduction and fattening in the same birds. While both pinealectomy and LL 300 lux treatments abolished the circadian rhythm of motor activity, they had no effect on the circannual rhythms of gonadal size and fattening. If the endogenous circadian rhythm in perch-hopping can be taken to reflect the circadian clock mechanism associated with gonadal functioning, present results suggest that circannual rhythm of reproduction in spotted munia is independent of circadian events.     

Circadian Rhythm of Blood Ethanol Clearance Rates in Rats: Response to Reversal of the L/D Regimen and to Continuous Darkness and Continuous Illumination

Phase relationships of the circadian rhythms of blood ethanol clearance (metabolic) rates and body temperature were studied in rats successively exposed to 4 illumination regimens: LD (light from 0800-2000 hr), DL (light from 2000-0800 hr), constant darkness (DD) and, lastly, constant light (LL). After a 4-wk standardization to each regimen, body temperatures were taken at 9 × 4-hr intervals to establish baseline circadian profiles. One week later, groups (N = 8) received 1.5 g/kg ethanol (i.p.) at 6 equally spaced timepoints during a 24-hr span, when temperatures were again measured. Ethanol clearance rates were estimated from decreasing blood ethanol levels sampled every 20 min from 60-200 min after dosing, and the resultant elimination curves were subjected to cosinor analysis. These studies show for the first time that the high amplitude circadian rhythm in ethanol metabolism persists under constant conditions of illumination (DD and LL), demonstrating that it may well be a truly internal circadian rhythm and not a response to exogenous cues of the light/dark cycle. During both LD and DL, maximal and minimal ethanol clearance rates fell near the end of the dark and light phases, respectively, and followed circadian peak and trough control temperatures by approximately 6 hr. A fixed internal phase relationship between the core body temperature and the circadian rhythm in ethanol metabolism is demonstrated, thus establishing the rhythm in body temperature as a suitable and convenient internal marker rhythm for studies of the metabolism of low-to-moderate ethanol doses. These studies demonstrate that the phase relationships of blood ethanol clearance rate and body temperature can be manipulated by the illumination regimen selected, an observation of both basic and practical importance.     

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.     

Circadian rhythms of body temperature and motor activity in rodents their relationships with the light-dark cycle

In rodents, the alternation of light and dark is the main synchronizer of circadian rhythms. The entrainment abilities of the LD cycle could be estimated by experimental modifications of the photoperiod and by following the subsequent temporal distribution of a circadian rhythm. The rate of reentrainment of a rhythm is determined by the nature of the studied variable, by the direction (advance or delay) and the magnitude (or value) of the phase shift. In rodents, core body temperature and motor activity are known to be well synchronized with each other under L:D 12:12 and under constant conditions (LL or DD). There are clear evidences that the circadian pattern of motor activity is generated by two oscillators, one from dusk signal and the other from dawn signal. Whether the circadian rhythms of body temperature and motor activity are generated by a common circadian mechanism or controlled by separate ones still remains unknown. The purpose of this review is to summarize the results obtained on the circadian rhythms of body temperature and motor activity throughout the daily cycle in order to clarify the relationships between these two functions.     

Modification of the hypothermic circadian cycles induced by DSIP and melatonin in pinealectomized and hypophysectomized rats

Both melatonin and DSIP (a nine amino acid peptide) effects have been previously shown to be (a) circadian rhythm related and (b) involved in inducing hypothermic effects in rats. In this study we report the hypothermia effects by each of these drugs alone and in combination when studied in normal (unoperated), pinealectomized, and hypophysectomized rats at various time points of the corresponding circadian cycle. A clear differential effect of drugs × time × preparation was found. While both DSIP and melatonin hypothermic effects were both circadian cycle dependent in intact rats the rhythmicity of melatonin hypothermic effect in pinealectomized rats, and DSIP hypothermic effect in hypophysectomized rats was missing. Although several hypotheses have been offered to account for the physiological mechanism(s) that govern the effects of the drugs, it is not yet possible to reliably relate the findings to existing neurochemical theory.     

Pinealectomy and LL Abolished Circadian Perching Rhythms but Did Not Alter Circannual Reproductive or Fattening Rhythms in Finches

In the subtropical finch, spotted munia (Lonchura punctulata) circannual rhythms (of gonads, fattening, feeding) have been demonstrated in an information-free environment of continuous illumination (LL), rendering it an ideal model for research on the physiology of the circannual clock. In an attempt to understand the involvement, if any, of the circadian system in the genesis of circannual rhythms, we studied the effect of pinealectomy (LL 15 lux) and strong continuous illumination (LL 300 lux), both known to abolish circadian rhythms, on the circadian perch-hopping rhythm and on the circannual rhythm of reproduction and fattening in the same birds. While both pinealectomy and LL 300 lux treatments abolished the circadian rhythm of motor activity, they had no effect on the circannual rhythms of gonadal size and fattening. If the endogenous circadian rhythm in perch-hopping can be taken to reflect the circadian clock mechanism associated with gonadal functioning, present results suggest that circannual rhythm of reproduction in spotted munia is independent of circadian events.     

Aschoff's rule in retinally degenerate mice

Both retinally degenerate and wildtype mice lengthened the period of their free-running circadian rhythms and reduced the amount of wheel running when exposed to increasing levels of constant illumination, in accordance with Aschoff's rule. Decreased locomotor activity may contribute toward lengthening of period in bright light. However, the known effects of activity on free-running period are small compared to those obtained by changing illumination. This suggests that Aschoff's rule in mice is not dependent on changes in nonphotic input, but results from a direct effect of light on the circadian system. The sparing of Aschoff's rule in retinally degenerate mice is further evidence that circadian photoreception depends on mechanisms other than rods and cones.     

Circadian rhythms of finches under steadily changing light intensity: Are self-sustaining circadian rhythms self-excitatory?

Summary Finches (Chloris chloris, Fringilla montifringilla) showed clear freerunning circadian rhythms when exposed to constant dim light. Increasing the light intensity by doubling it each day made them become arrhythmic at a certain threshold intensity of illumination, showing continuous locomotor activity. When the light intensity was decreased steadily at the reversed rate, the finches became rhythmic again. 7 out of 8 finches had a clear start in their rhythms, from one day to the next, at light intensities about 4 times higher than the point where they had become arrhythmic. The last finch started its freerunning circadian rhythm gradually, a few days after the light intensity had reached a constant dim illumination (0.2 lux).The results of all birds are taken as proof of the self-excitatory capacity of the circadian system. This means, it characterizes the dynamics of the system that the clock mechanism is continuously in operation, and not only after a passive reaction to external stimuli exceeds any threshold. Simultaneously, the results of all but one bird allow the evaluation of the contribution of proportional and differential effects of light in the control of circadian rhythmicity. A relative change in light intensity by 100% in the course of one day is nearly equivalent to a change of 100% in the absolute intensity of illumination.     

Locomotor Rhythms in the Fiddler Crab, Uca subcylindrica

The locomotor activities of individual specimens of Uca subcylindrica (Stimpson) collected from semi-arid, supratidal habitats in south Texas and northeastern Mexico were studied in the laboratory using periodogram analysis. When crabs were placed under constant darkness (DD) or constant illumination (LL), free-running circadian rhythms were observed in the activity recordings. The locomotor activity of strongly rhythmic crabs in LL has an average period length of 24.4 h. Crabs held in DD express motor rhythms with periods of approximately 24.0 h. In LL the most common wave form for activity is unimodal, while under DD it is bimodal. Recordings under natural illumination (NL) revealed that both period length and the time of maximum activity (phasing) varied through the year. During winter months, the crabs are primarily diurnal with peaks in activity occurring between 0900 and 2100 h and possess a circadian rhythm with a 23.9 h period. During summer, crabs were nocturnal with maximal activity between 1300 and 0600 and a circadian period closer to 24.0 h. In these experiments, the rhythmic locomotor activities of U. subcylindrica are best described as “circadian”. This is unusual for a genus known for its expression of circatidal and circalunidian rhythms.