Delta sleep-inducing peptide (DSIP): a still unresolved riddle

Delta sleep-inducing peptide (DSIP) was isolated from rabbit cerebral venous blood by Schoenenberger-Monnier group from Basel in 1977 and initially regarded as a candidate sleep-promoting factor. However, the link between DSIP and sleep has never been further characterized, in part because of the lack of isolation of the DSIP gene, protein and possible related receptor. Thus the hypothesis regarding DSIP as a sleep factor is extremely poorly documented and still weak. Although DSIP itself presented a focus of study for a number of researchers, its natural occurrence and biological activity still remains obscure. DSIP structure is different from any other known representative of the various peptide families. In this mini-review we hypothesize the existence of a DSIP-like peptide(s) that is responsible (at least partly) for DSIP-like immunoreactivity and DSIP biological activity. This assumption is based on: (i) a highly specific distribution of DSIP-like immunoreactivity in the neurosecretory hypothalamic nuclei of various vertebrate species that are not particularly relevant for sleep regulation, as revealed by the histochemical studies of the Geneva group (Charnay et al.); (ii) a large spectrum of DSIP biological activity revealed by biochemical and physiological studies in vitro; (iii) significant slow-wave sleep (SWS) promoting activity of certain artificial DSIP structural analogues (but not DSIP itself!) in rabbits and rats revealed by our early studies; and (iv) significant SWS-promoting activity of a naturally occurring dermorphin-decapeptide that is structurally similar to DSIP (in five of the nine positions) and the sleep-suppressing effect of its optical isomer, as revealed in rabbits. Potential future studies are outlined, including natural synthesis and release of this DSIP-like peptide and its role in neuroendocrine regulation.     

Aminopeptidase in human CSF which degrades delta-sleep inducing peptide (DSIP)

This study describes the purification and characterization of an aminopeptidase from human cerebrospinal fluid capable of degrading delta-sleep-inducing-peptide (DSIP). The enzyme has an apparent molecular weight of approximately 80,000 dalton. It is sensitive towards amastatin, bestatin and EDTA and is optimally active at neutral pH. The recovered enzyme was also found to degrade other neuropeptides, e.g., the enkephalins.     

Delta sleep-inducing peptide (DSIP)-like immunoreactivity in gut: coexistence with known peptide hormones

Delta sleep-inducing peptide-like immunoreactivity (DSIP-LI) has previously been demonstrated in brain neurons and in endocrine cells of the pituitary and the adrenal medulla. By means of three different antisera against synthetic DSIP we now describe the occurrence and distribution of DSIP-LI in several gut endocrine cells. The human gut was the richest source, where DSIP-LI was located in gastrin/CCK, secretin and PYY/glicentin cells. The rat and pig gut harbour a moderate number of immunoreactive cells in the antral mucosa but in the intestines DSIP-LI-containing cells were very few. By radioimmunoassay, the concentration of DSIP-LI was determined in extracts of various gut regions from man, pig and rat. The highest concentrations were found in all human specimens compared with corresponding samples in the pig and rat. In all three species, high-performance liquid chromatography revealed a single peak of DSIP-like material with approximately the same retention time as DSIP 3-9. Taken together, the present results provide evidence for the presence of DSIP-LI in gut endocrine cells in man, pig and rat; the human gut seems to be the richest source of DSIP-like peptides.     

Delta sleep-inducing peptide (DSIP) stimulates growth hormone (GH) release in the rat by hypothalamic and pituitary actions

To evaluate possible effects of delta sleep-inducing peptide on GH release, the peptide was micro-injected into conscious animals with third ventricular cannulae and blood samples were drawn from indwelling external jugular vein cannulae. Ovariectomized animals were used in order to eliminate gonadal steroid feedback. In the initial experiment, intraventricular injection of 5 micrograms of the peptide induced an elevation of GH which became significant by 30 min and persisted for the 120 min duration of the experiment after the injection. Diluent-injected animals showed a slight initial drop in GH and then no increase. The increase in plasma GH induced by the peptide was dose-related with a minimal effective dose of 0.1 microgram and a linear log-dose increase to a dose of 10 micrograms. This effect is presumably mediated hypothalamically via a dopaminergic mechanism since it could be blocked by pre-treatment of the animals with pimozide, a dopamine receptor blocker. Dispersed overnight, cultured pituitary cells from ovariectomized rats exhibited a dose-related increase in GH release in static incubations with DSIP. A response occurred with the lowest dose tested (10(-12) M) which increased to a maximum at 10(-10) M DSIP. The responses then declined at higher doses such that they were no longer significant at doses of 10(-7) and 10(-5) M. The increase even at the most effective dose was approximately 50% above the basal values. The results are consistent with the hypothesis that DSIP may be involved in GH release via a dopaminergic mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)     

Delta sleep-inducing peptide (DSIP) stimulates LH release in steroid-primed ovariectomized rats

Delta sleep inducing peptide (DSIP) has been shown to increase sleep in various animals and it is found in various parts of the brain including the hypothalamus. While intraventricular administration of DSIP (2 or 10 micrograms) failed to affect LH release in ovariectomized rats, in two separate experiments DSIP (2 or 10; 15 or 30 micrograms) promptly stimulated LH release in ovariectomized estrogen, progesterone-primed rats. However, DSIP (10(-8) or 10(-6)M) had no effect on either basal or luteinizing hormone-releasing hormone-induced in vitro LH release from the hemipituitaries of ovarian steroid-primed rats. These findings are in accord with the hypothesis that DSIP or DSIP-like peptide(s) may activate the hypothalamic neural circuitry responsible for stimulation of LH release reported to occur during sleep.     

Effect of mitochondria and microsomal fractions on glycolytic activity of rat brain and liver tissues under hypoxia

It is found that acute hypoxia inhibits the glycolytic activity of postmitochondrial fraction in the liver, activates in the brain, but has no effect on glycolysis under conditions of a preliminary administration of diethylaminoethylamide of parachlorophenoxyacetic acid--antihypoxic preparation. In the processes of two- and four-week interrupted training of adaptation to hypoxia the activity of the liver glycolytic system rises. Suspensions of the mitochondric and microsomal fraction added in definite ratios to the postmitochondrial fraction of the brain and liver intensify its glycolytic activity both in control and hypoxic animals. The activating effect of mitochondria is higher as compared with the control when glycolysis is decreased; when glycolysis is increased the phenomenon is not observed. A mechanism of the found changes in glycolysis and the validity of the tissue glycolysis estimation from the activity of the postmitochondrial fraction are discussed.     

Delta sleep-inducing peptide (DSIP)-like material is absorbed by the gastrointestinal tract of the neonatal rat

Entry of delta sleep-inducing peptide (DSIP) into the circulation from the gastrointestinal (GI) tract was studied in unweaned rat pups. The pups were fed an analog of DSIP (N-Tyr-DSIP) or 125I-N-Tyr-DSIP and blood samples collected. Significant increases in plasma DSIP-like immunoreactivity occurred after the feeding of 100 ωg/animal of N-Tyr-DSIP but not after vehicle (normal saline) or 1 ωg/animal. Column chromatography showed this immunoreactivity to coelute with intact DSIP and des-Trp¹- DSIP. A small but statistically significant increase of immunoreactivity occurred in the plasma of pups whose nursing mothers were injected with N-Tyr-DSIP but not in those whose mothers were injected with saline. Radioactivity appeared in both the brain and blood of 1–2 and 10 day old rat pups fed 125I-N-Tyr-DSIP. Although only a small amount of the radioactivity in plasma co-eluted with intact 1251I-N-Tyr-DSIP on column chromatography, almost all of the radioactivity in brain did, suggesting that the radioactivity in the brain represented crossing of the blood-brain-barrier by the peptide and not just contamination by blood. The results cannot be explained by either regurgitation of intestinal contents, or by stimulation of endogenous peptide. They show that a DSIP peptide administered orally can be absorbed through the GI tract into the systemic circulation.     

Activity-dependent neuroprotective protein (ADNP)-derived peptide (NAP) ameliorates hypobaric hypoxia induced oxidative stress in rat brain

Hypobaric hypoxia is a socio-economic problem affecting cognitive, memory and behavior functions. Severe oxidative stress caused by hypobaric hypoxia adversely affects brain areas like cortex, hippocampus, basal ganglia, and cerebellum. In the present study, we have investigated the antioxidant and memory protection efficacy of the synthetic NAP peptide (NAPVSIPQ) during long-term chronic hypobaric hypoxia (7, 14, 21 and 28 days, 25,000 ft) in rats. Intranasal supplementation of NAP peptide (2 μg/Kg body weight) improved antioxidant status of brain evaluated by biochemical assays for free radical estimation, lipid peroxidation, GSH and GSSG level. Analysis of expression levels of SOD revealed that NAP significantly activated antioxidant genes as compared to hypoxia exposed rats. We have also observed a significant increased expression of Nrf2, the master regulator of antioxidant defense system and its downstream targets such as HO-1, GST and SOD1 by NAP supplementation, suggesting activation of Nrf2-mediated antioxidant defense response. In corroboration, our results also demonstrate that NAP supplementation improved the memory function assessed with radial arm maze. These cumulative results suggest the therapeutic potential of NAP peptide for ameliorating hypobaric hypoxia-induced oxidative stress.     

Transport of malate and citrate into rat brain mitochondria under hypoxia and anesthesia

Hypoxia and anesthesia inhibited penetration of malate and citrate into the brain mitochondria by 60% and 40%, respectively. Anesthetized animals exposed to low oxygen tension showed similar changes as those subjected to hypoxia without anesthesia. Recovery from anesthesia was rapid and the rates of citrate and malate uptake returned to the control values in approximately 60 min. In the case of hypoxia, days were required to restore the control values of citrate and malate uptake. Free fatty acids had no effect on the entry of malate and citrate into the mitochondria. However, changes in the levels of protein—SH groups were observed which may be responsible for the impaired transport of citrate and malate under hypoxic condition.     

Delta sleep-inducing peptide (DSIP)-like material exists in peripheral organs of rats in large dissociable forms

The presence of delta sleep-inducing peptide (DSIP) in brain has been shown by radioimmunoassay (RIA) and by immunocytochemistry. We now describe the occurrence of DSIP-like material in the peripheral organs of the rat as measured by RIA. Tissue from 12 areas was extracted with water, and the amounts of immunoreactive material found to be between 86 pg/mg tissue (muscle) and 849 pg/mg (stomach). Recoveries of about 80% of added DSIP were achieved at tissue concentrations of 1 mg/ml or less. This percentage was reduced in liver at higher concentrations. The percentage of small peptide adsorbed by charcoal was greatly increased at lower tissue concentrations in all organs. This effect was significant and linear. Chromatography on columns of Sephadex G-15 and G-25 showed immunoreactive material mostly larger than DSIP. Digestion with trypsin, however, produced small immunoreactive peptides with only a minimal reduction in total immunoreactivity. Thus, DSIP-like material is widespread in peripheral tissues and appears to exist mainly in a large form, probably bound to protein, that can be reduced in size by tryptic digestion and can be dissociated at lower concentrations of tissue to yield small immunoreactive peptides.     

Delta-sleep inducing peptide (DSIP) and its fragments as the modulators of neural transmission in the central nervous system

The present study is a continuation of our previous experiments on DSIP activity which have revealed that nonapeptide DSIP inhibits hippocampal electrical activity of the 4-7 c/s frequency band. The aim of the present study was to find which of the known DSIP fragments is responsible for its activity, i.e. to find the active site of the molecule. The experiments were carried out with the entire DSIP molecule and its three different fragments. The method of threshold continuous arousal pattern (TCAP) monitoring was used as the indicator of DSIP activity. It was found that the entire DSIP molecule increased TCAP, while its 1-5 fragment decreased it 1-4 and 5-9 fragments had no noticeable effect.     

Effect of a short-term fasting and altitude hypoxia on the cytochrome oxidase activity of rat brain mitochondria

The effect of short-term fasting and thirst, prolonged fasting and hypoxic hypoxia upon the activity of cytochrome oxidase was studied in mitochondrial fractions obtained from the brain and the liver. The investigation was carried out in two groups of rats, 5 and 60 days old. a) The activity of cytochrome oxidase in mitochondria isolated from the brain cortex, subcortical regions and the medulla oblongata rises, while the changes in liver mitochondrial fractions are reverse. b) A significant increase of mitochondrial cytochrome oxidase was found in 5-day-old rats after both types of fasting and hypoxia in all regions of the brain, as well as in the liver. c) The cytochrome oxidase activity in brain and liver mitochondria of 60-day-old rats was not affected appreciably after 24 h nutritional deprivation, with the exception of a significant rise of activity in the medulla oblongata. Prolonged fasting and hypoxia again markedly increased the activity of this enzyme in all regions of the brain and in the liver.     

Rapid eye movement (rem) sleep deprivation: effect on acid mucopolysaccharides in rat brain.

The effect of rapid eye movement (REM) sleep deprivation on the total content and proportion of different mucopolysaccharides (AMPS) containing uronic acid in rat brain was studied. REM sleep deprivation was induced by the water tank methods. Five experimental groups of animals were used: control, stressed, REM sleep deprived, post-stress sleeping and post-deprivation sleeping rats. No changes of AMPS were observed in any of the experimental groups when the whole brain was analysed. A significant increase of AMPS was found in the cerebral hemispheres of stressed and REM deprived rats. A significant decrease of AMPS was observed in the cerebellum and brain stem. A further increase of AMPS was found in the cerebral hemispheres after the rebound of REM sleep following its deprivation, and after the recovery sleep following the stress. A significant increase of AMPS was found in the brain stem of rats allowed to recuperate after REM deprivation or stress as compared with the stressed and REM deprived animals. Recovery sleep induced a significant increase of AMPS in the cerebellum in previously stressed rats, while previously REM deprived rats exhibited a further decrease of AMPS from control values. The possible functional meaning of these results is discussed in relation to the role of REM sleep in protein synthesis and learning and memory processes. Intriguing, well-controlled positive findings and the fact that no experimental design is known where stress is minimal while REM deprivation is 100 per cent, justify and encourage continued efforts in studying the biochemical state of the brain during sleep and/or its alterations.     

Level of delta sleep-inducing peptide (DSIP) in the brain of rats with different alcoholic motivation

Radioimmunoassay was used to measure the content of delta-sleep-inducing peptide (DSIP) in random-bred albino rats divided into groups according to the duration of ethanol anesthesia and the levels of 15% ethanol consumption under free-choice conditions. The concentration of the neuropeptide was assayed in intact brain, in the cortex of large hemispheres, medulla oblongata, thalamus and striatum. The short-sleeping rats manifested a statistically significant lowering of the DSIP content in intact brain homogenates, in the cortex of large hemispheres and striatum. On the contrary, thirty minutes after a single intraperitoneal injection of ethanol in a dose of 1 g/kg the DSIP content in the medulla oblongata, thalamus and striatum was found to be increased. The raising of the ethanol dose up to 2.5 and 4.5 g/kg was followed by a less significant increase in the neuropeptide content. Prolonged chronic alcoholization under free-choice conditions led after 12 months to the reduced DSIP content in the medulla oblongata, thalamus and striatum. The importance of DSIP for the pathogenesis of experimental alcoholism using rats with different levels of alcoholic motivation is discussed.     

Delta-sleep-inducing peptide (DSIP) inhibited CRF-induced ACTH secretion from rat anterior pituitary gland in vitro

Delta-sleep-inducing peptide (DSIP, 10(-9) - 10(-7) M) significantly inhibited the CRF-induced ACTH release from rat anterior pituitary quarters in vitro. 10(-8) M DSIP showed the most prominent inhibition. DSIP (10(-8) M) also inhibited the CRF-activated cAMP levels in anterior pituitary tissue. DSIP did not influence basal ACTH or cAMP levels. Prostaglandin E2 (PGE2)-release from anterior pituitary quarters was not changed by DSIP. From these results, we conclude that DSIP inhibits CRF-induced ACTH release at the pituitary level through the inhibition of the cAMP system in corticotrophs. The involvement of PGE2 in this phenomenon is unlikely.