Measuring activity-rest rhythms under different acclimation periods in a marine fish using automatic deep learning-based video tracking.

Most organisms synchronize to an approximately 24-hour (circadian) rhythm. This study introduces a novel deep learning-powered video tracking method to assess the stability, fragmentation, robustness and synchronization of activity rhythms in Xyrichtys novacula. Experimental X. novacula were distributed into three groups and monitored for synchronization to a 14/10 hours of light/dark to assess acclimation to laboratory conditions. Group GP7 acclimated for 1 week and was tested from days 7 to 14, GP14 acclimated for 14 days and was tested from days 14 to 21 and GP21 acclimated for 21 days and was tested from days 21 to 28. Telemetry data from individuals in the wild depicted their natural behavior. Wild fish displayed a robust and minimally fragmented rhythm, entrained to the natural photoperiod. Under laboratory conditions, differences in activity levels were observed between light and dark phases. However, no differences were observed in activity rhythm metrics among laboratory groups related to acclimation period. Notably, longer acclimation (GP14 and GP21) led to a larger proportion of individuals displaying rhythm synchronization with the imposed photoperiod. Our work introduces a novel approach for monitoring biological rhythms in laboratory conditions, employing a specifically engineered video tracking system based on deep learning, adaptable for other species.

Activity-Rest Circadian Rhythm of the Pearly Razorfish in Its Natural Habitat, before and during Its Mating.

Recent technological advances in marine biotelemetry have demonstrated that marine fish species perform activity-rest rhythms that have relevant ecological and evolutionary consequences. The main objective of the present report is to study the circadian rhythm of activity-rest of the pearly razorfish, Xyrichtys novacula in its own habitat, before and during the reproduction season using a novel biotelemetry system. This fish species is a small-bodied marine species that inhabits most shallow soft habitats of temperate areas and has a high interest for commercial and recreational fisheries. The activity of free-living fish was monitored by means of high-resolution acoustic tracking of the motor activity of the fish in one-minute intervals. The obtained data allowed the definition of the circadian rhythm of activity-rest in terms of classical non-parametric values: interdaily stability (IS), intradaily variability (IV), relative amplitude (RA), average activity during the most-active period of consecutive 10 h (M10), and average activity during the least-active period of consecutive 5 h (L5). We observed a well-marked rhythm, with little fragmentation and good synchrony with the environmental cycle of light-darkness, regardless of sex and the period studied. However, the rhythm was found to be slightly more desynchronized and fragmented during reproduction because of variations in the photoperiod. In addition, we found that the activity of the males was much higher than that of the females (p < 0.001), probably due to the peculiar behavior of the males in defending the harems they lead. Finally, the time at which activity began in males was slightly earlier than it was in females (p < 0.001), presumably due to the same fact, as differences in activity or for the individual heterogeneity of this species in the time of awakening are considered to be an independent axis of the fish's personality. Our work is novel, as it is one of the first studies of activity-rest rhythm using classical circadian-related descriptors in free-living marine fish using locomotory data facilitated by novel technological approaches.

Mammalian NREM and REM sleep: Why, when and how.

This report proposes that fish use the spinal-rhombencephalic regions of their brain to support their activities while awake. Instead, the brainstem-diencephalic regions support the wakefulness in amphibians and reptiles. Lastly, mammals developed the telencephalic cortex to attain the highest degree of wakefulness, the cortical wakefulness. However, a paralyzed form of spinal-rhombencephalic wakefulness remains in mammals in the form of REMS, whose phasic signs are highly efficient in promoting maternal care to mammalian litter. Therefore, the phasic REMS is highly adaptive. However, their importance is low for singletons, in which it is a neutral trait, devoid of adaptive value for adults, and is mal-adaptive for marine mammals. Therefore, they lost it. The spinal-rhombencephalic and cortical wakeful states disregard the homeostasis: animals only attend their most immediate needs: foraging defense and reproduction. However, these activities generate allostatic loads that must be recovered during NREMS, that is a paralyzed form of the amphibian-reptilian subcortical wakefulness. Regarding the regulation of tonic REMS, it depends on a hypothalamic switch. Instead, the phasic REMS depends on an independent proportional pontine control.

The Birth of the Mammalian Sleep.

Mammals evolved from small-sized reptiles that developed endothermic metabolism. This allowed filling the nocturnal niche. They traded-off visual acuity for sensitivity but became defenseless against the dangerous daylight. To avoid such danger, they rested with closed eyes in lightproof burrows during light-time. This was the birth of the mammalian sleep, the main finding of this report. Improved audition and olfaction counterweighed the visual impairments and facilitated the cortical development. This process is called "The Nocturnal Evolutionary Bottleneck". Pre-mammals were nocturnal until the Cretacic-Paleogene extinction of dinosaurs. Some early mammals returned to diurnal activity, and this allowed the high variability in sleeping patterns observed today. The traits of Waking Idleness are almost identical to those of behavioral sleep, including homeostatic regulation. This is another important finding of this report. In summary, behavioral sleep seems to be an upgrade of Waking Idleness Indeed, the trait that never fails to show is quiescence. We conclude that the main function of sleep consists in guaranteeing it during a part of the daily cycle.

Comparing the Behavioural Effects of Exogenous Growth Hormone and Melatonin in Young and Old Wistar Rats.

Growth hormone (GH) and melatonin are two hormones with quite different physiological effects. Curiously, their secretion shows parallel and severe age-related reductions. This has promoted many reports for studying the therapeutic supplementation of both hormones in an attempt to avoid or delay the physical, physiological, and psychological decay observed in aged humans and in experimental animals. Interestingly, the effects of the external administration of low doses of GH and of melatonin were surprisingly similar, as both hormones caused significant improvements in the functional capabilities of aged subjects. The present report aims at discerning the eventual difference between cognitive and motor effects of the two hormones when administered to young and aged Wistar rats. The effects were tested in the radial maze, a test highly sensitive to the age-related impairments in working memory and also in the rotarod test, for evaluating the motor coordination. The results showed that both hormones caused clear improvements in both tasks. However, while GH improved the cognitive capacity and, most importantly, the physical stamina, the effects of melatonin should be attributed to its antioxidant, anxiolytic, and neuroprotective properties.

Cognitive recovery and restoration of cell proliferation in the dentate gyrus in the 5XFAD transgenic mice model of Alzheimer's disease following 2-hydroxy-DHA treatment.

Alzheimer's disease (AD) is the most common neurodegenerative disorder in the elderly. In the last years, abnormalities of lipid metabolism and in particular of docosahexaenoic acid (DHA) have been recently linked with the development of the disease. According to the recent studies showing how hydroxylation of fatty acids enhances their biological activity, here we show that chronic treatment with a hydroxylated derivative of DHA, the 2-hydroxy-DHA (2OHDHA) in the 5XFAD transgenic mice model of AD improves performance in the radial arm maze test and restores cell proliferation in the dentate gyrus, with no changes in the presence of beta amyloid (Aß) plaques. These results suggest that 2OHDHA induced restoration of cell proliferation can be regarded as a major component in memory recovery that is independent of Aß load thus, setting the starting point for the development of a new drug for the treatment of AD.

Asymmetric sleep in rats.

Five Wistar rats were surgically implanted with cortical and parietal electrodes for conventional polysomnography to test for sleep-related EEG asymmetries during 48 hours of continuous recording. When the animals were grouped not according to right-left dominance (which would represent a population bias) but instead according to preferred vs non-preferred hemisphere, significant light/dark circadian changes in side dominance were found in delta power during NREM; in theta and beta power during REM; and in alpha 1, alpha 2, and theta power during wakefulness. The changes have been interpreted as a response to temporal variations in the capability to respond to environmental challenges.

Evolution of wakefulness, sleep and hibernation: from reptiles to mammals.

Thus far, most hypotheses on the evolutionary origin of sleep only addressed the probable origin of its main states, REM and NREM. Our article presents the origin of the whole continuum of mammalian vigilance states including waking, sleep and hibernation and the causes of the alternation NREM-REM in a sleeping episode. We propose: (1) the active state of reptiles is a form of subcortical waking, without homology with the cortical waking of mammals; (2) reptilian waking gave origin to mammalian sleep; (3) reptilian basking behaviour evolved into NREM; (4) post-basking risk assessment behaviour, with motor suspension, head dipping movements, eye scanning and stretch attending postures, evolved into phasic REM; (5) post-basking, goal directed behaviour evolved into tonic REM and (6) nocturnal rest evolved to shallow torpor. A small number of changes from previous reptilian stages explain these transformations.

Melatonin and tryptophan affect the activity-rest rhythm, core and peripheral temperatures, and interleukin levels in the ringdove: changes with age.

Aging is known to alter the circadian rhythms of melatonin, serotonin, thermoregulatory responses, cytokine production, and sleep/wakefulness which affect sleep quality. We tested the possible palliative effects of a 3-day administration of melatonin (0.25 or 2.5 mg/kg of body weight [b.w.] to young and old ringdoves, respectively) or tryptophan (300 mg/kg of b.w. to old ringdoves) on these rhythms. Doves are a monophasic, diurnal species; these characteristics are similar in humans. Old animals presented lower melatonin and serotonin levels; higher interleukin (IL)-1beta, IL-6, and tumor necrosis factor alpha values; and reductions in the Midline-Estimating Statistic of Rhythm and amplitude of activity-rest rhythm and in the amplitude of the core temperature rhythm. Melatonin raised serum melatonin levels; tryptophan increased both melatonin and serotonin levels. Melatonin and tryptophan lowered nocturnal activity, core temperature, and cytokine levels and increased peripheral temperature in both groups. Melatonin or tryptophan may limit or reverse some of the changes that occur in sleep-wake rhythms and temperature due to age.

Chrononutrition: use of dissociated day/night infant milk formulas to improve the development of the wake-sleep rhythms. Effects of tryptophan.

Three different lactation experiments have been tested in a double blind procedure for 3 weeks, to improve sleep-wake patterns in infants. In a control experiment, standard infant commercial milk (1.5% tryptophan) was administered without changes during the day. In a second control (inverse), enriched milk (3.4% tryptophan) was given during light-time (06.00-18.00h), and standard commercial milk during night-time (18.00-06.00h). During the experimental week, the infants received standard milk during light-time and tryptophan enriched milk during night-time. The infants receiving the enriched formula during dark time showed improvements in the sleep parameters studied, and no statistical differences were found between the two control lactations. The urinary metabolites of serotonin suggest that the observed improvements were due to an increased use of serotonin to melatonin synthesis. In conclusion, the chronobiological changes in the normal components of the diet can improve infantile development of sleep/wake rhythms.

Opposite effects of tryptophan intake on motor activity in ring doves (diurnal) and rats (nocturnal).

The role of l-tryptophan as precursor of serotonin and melatonin synthesis on activity-rest rhythm was studied in ring doves, Streptopelia risoria, as a representative of diurnal animals and rats, Rattus norvegicus, as a typical nocturnal one. The animals were housed in cages equipped for horizontal activity recording in a thermostatized chamber and submitted to a 12/12h light/dark photoperiod (lights on at 08:00 h). After acclimatization, the animals received vehicle (methylcellulose) and l-tryptophan (240 mg/kg) by esophagic cannula 2h before the onset of either light or dark phase. Also, oral melatonin (2.5mg/kg) was tested for comparative purposes. After nocturnal l-tryptophan administration, rats showed increased activity (149%), while the opposite occurred in ring doves (39% decrease). No significant changes were found after diurnal l-tryptophan intake in either species. Melatonin produced effects similar to those of l-tryptophan. These results suggest that the effects of l-tryptophan administration are dependent on the nocturnal/diurnal habits of the studied species and, most probably, are mediated by increased melatonin synthesis.

Age related changes in the activity-rest circadian rhythms and c-fos expression of ring doves with aging. Effects of tryptophan intake.

Age related changes in the circadian rhythms and sleep quality has been linked with impairment in the function of the suprachiasmatic nucleus (SCN) and melatonin secretion. The precursor of melatonin, serotonin (5-HT) is a neurotransmitter involved in the synchronisation of the circadian clock located in SCN, which shows decreased levels with age. The present work studied the effects of L-tryptophan, the precursor of 5-HT, on the circadian activity-rest rhythm and c-fos expression in the SCN of young and old ring doves, animals diurnal and monocyclic as humans. Two hours before the onset of dark phase, animals housed in cages equipped for activity recording and maintained under 12/12 L/D conditions, received orally L-tryptophan (100 and 240 mg/kg) and, for comparative purposes, melatonin (2.5 and 5 mg/kg). The administration of both L-tryptophan and melatonin reduced the nocturnal activity of all ring doves although only the highest doses were effective in old ones. A reduced amplitude in the activity-rest rhythm was observed in old animals in comparison to youngest, but it was increased after the treatments. Sleep parameters, calculated from the activity data, indicated a worsened sleep quality in old animals but it was improved with the treatments. In addition, the expression of c-fos in the SCN was reduced after both mentioned treatments. The results point to the SCN as a target for the observed nocturnal effects of L-tryptophan and melatonin, and support the supplemental administration of the essential amino acid L-tryptophan to reverse the disturbances of the circadian activity-rest cycle related with ageing.

Age-related changes in circadian rhythm of serotonin synthesis in ring doves: effects of increased tryptophan ingestion.

Alterations in the function of the hypothalamic suprachiasmatic nucleus (SCN) with age have been reported. As serotonin is an important regulator of the circadian clock located in SCN, this work studied the changes produced in the synthesis of serotonin with age using the accumulation of 5-HTP after decarboxylase inhibition as a measure of serotonin synthesis in the brain in vivo, in young and old ring doves at the onset of lights-on and lights-off. A diurnal cycle in tryptophan hydroxylation was observed in young animals, with an increased daylight synthesis and metabolism of 5-HT in hippocampus, neostriatum and hypothalamus. A single dose of melatonin (1 mg/kg, i.p., 1 h) at lighttime produced an inhibitory effect on the synthesis of 5-HT. In contrast, differences in 5-HT synthesis and metabolism between day and night disappeared in old animals indicating an absence of a circadian rhythm in 5-HT synthesis and metabolism. The administration of L-tryptophan (240 mg/kg, i.p.) strongly increased the 5-HT synthesis in young animals only during lights-off time while it increased in old ones irrespective of the administration time. These results suggest that the supplemental administration of tryptophan might aid to improve the descent in 5-HT that normally occurs, as animals get old.

Mammalian sleep may have no adaptive advantage over simple activity-rest cycles.

The adaptive value of sleep remains unknown in spite of the intense research performed throughout the last decades. However, few sleep researchers are aware of the difficulties posed by the blind acceptance of an extreme adaptationist viewpoint. Under this philosophy, every anatomical and functional detail present in a living being should have a positive adaptive value, a position that has been considered as rather doubtful. In this report, it is proposed that most of the physiological changes used for mammalian sleep definition could be mere by-products of other true adaptations, such as the ontogenetic and phylogenetic development of the nervous system. As a result, complex mammalian sleep could have no adaptive value over that of the simplest forms of rest-activity cycles present in all living forms. In addition, it is proposed that the absence of adaptive value should, by default, be the first option regarding the function of sleep. Besides, the burden of the proof should be always charged over the proponents of every particular adaptive function. As this proof has not been reached, it is the absence of function for sleep which should be taken for granted.

Human Sleep Apneas and Animal Diving Reflexes: The Comparative Link.

Adaptations to survive periods of limited access to oxygen should have been favored along the evolution of vertebrates. Paradigmatic examples of this adaptation are the diving animals, which can sustain prolonged and repetitive periods of anoxia. These animals support what would be considered a severe gas imbalance in their internal environment thanks to three main strategies: increased oxygen stores, resistance to asphyxia, and reduced metabolic expenditure during the apneic intervals. However, diving animals developed their abilities from very old life-sustaining responses that should have been used on many other occasions. Humans with sleep apneas perhaps share many physiological adaptations with diving animals. We review here the extent of such similarities and offer clear evidence of its existence and suggest possible research lines that could improve the clinical knowledge about this condition.