Dopaminergic modulation of arousal in Drosophila

BACKGROUND: Arousal levels in the brain set thresholds for behavior, from simple to complex. The mechanistic underpinnings of the various phenomena comprising arousal, however, are still poorly understood. Drosophila behaviors have been studied that span different levels of arousal, from sleep to visual perception to psychostimulant responses. RESULTS: We have investigated neurobiological mechanisms of arousal in the Drosophila brain by a combined behavioral, genetic, pharmacological, and electrophysiological approach. Administration of methamphetamine (METH) suppresses sleep and promotes active wakefulness, whereas an inhibitor of dopamine synthesis promotes sleep. METH affects courtship behavior by increasing sexual arousal while decreasing successful sexual performance. Electrophysiological recordings from the medial protocerebrum of wild-type flies showed that METH ingestion has rapid and detrimental effects on a brain response associated with perception of visual stimuli. Recordings in genetically manipulated animals show that dopaminergic transmission is required for these responses and that visual-processing deficits caused by attenuated dopaminergic transmission can be rescued by METH. CONCLUSIONS: We show that changes in dopamine levels differentially affect arousal for behaviors of varying complexity. Complex behaviors, such as visual perception, degenerate when dopamine levels are either too high or too low, in accordance with the inverted-U hypothesis of dopamine action in the mammalian brain. Simpler behaviors, such as sleep and locomotion, show graded responses that follow changes in dopamine level.     

Re-patterning sleep architecture in Drosophila through gustatory perception and nutritional quality

Organisms perceive changes in their dietary environment and enact a suite of behavioral and metabolic adaptations that can impact motivational behavior, disease resistance, and longevity. However, the precise nature and mechanism of these dietary responses is not known. We have uncovered a novel link between dietary factors and sleep behavior in Drosophila melanogaster. Dietary sugar rapidly altered sleep behavior by modulating the number of sleep episodes during both the light and dark phase of the circadian period, independent of an intact circadian rhythm and without affecting total sleep, latency to sleep, or waking activity. The effect of sugar on sleep episode number was consistent with a change in arousal threshold for waking. Dietary protein had no significant effect on sleep or wakefulness. Gustatory perception of sugar was necessary and sufficient to increase the number of sleep episodes, and this effect was blocked by activation of bitter-sensing neurons. Further addition of sugar to the diet blocked the effects of sweet gustatory perception through a gustatory-independent mechanism. However, gustatory perception was not required for diet-induced fat accumulation, indicating that sleep and energy storage are mechanistically separable. We propose a two-component model where gustatory and metabolic cues interact to regulate sleep architecture in response to the quantity of sugar available from dietary sources. Reduced arousal threshold in response to low dietary availability may have evolved to provide increased responsiveness to cues associated with alternative nutrient-dense feeding sites. These results provide evidence that gustatory perception can alter arousal thresholds for sleep behavior in response to dietary cues and provide a mechanism by which organisms tune their behavior and physiology to environmental cues.     

Sleep state switching

We take for granted the ability to fall asleep or to snap out of sleep into wakefulness, but these changes in behavioral state require specific switching mechanisms in the brain that allow well-defined state transitions. In this review, we examine the basic circuitry underlying the regulation of sleep and wakefulness and discuss a theoretical framework wherein the interactions between reciprocal neuronal circuits enable relatively rapid and complete state transitions. We also review how homeostatic, circadian, and allostatic drives help regulate sleep state switching and discuss how breakdown of the switching mechanism may contribute to sleep disorders such as narcolepsy.     

A contextual definition of male sexual arousal

Sexual arousal is a construct without a widely shared definition. Historically, sexual arousal has usually referred to a central physiological state, but there has been much less agreement on its relation to motivation, emotion, and - for males - penile erection and ejaculation. Many behavioral and physiological measures have been used as operational definitions of sexual arousal, but the relation of the measure to arousal is often assumed rather than tested. For men, penile erection in the presence of erotic stimuli has been considered the most reliable and valid indicator of sexual arousal. The adoption of analogous criteria is recommended for research on other male mammals in order to establish a minimal basis for inferring that they are sexually aroused. That is, sexual arousal should be inferred only when penile erection is observed in a sexual context. A sexual context is provisionally defined as an environment that tends in most reproductively active males of the species to provoke further sexual stimulation, e.g., copulation or self-stimulation to ejaculation. Erection occurring outside of a sexual context, as during REM sleep or from injection of drugs, is not grounds for inferring arousal. Conversely, males engaging in behavior directed toward estrous females may be sexually motivated, but in the absence of erection, the males should not be assumed to be sexually aroused. Implications of other erection-context interactions are also considered. Adoption of these more conservative criteria for inferring sexual arousal may promote greater precision in identifying the physiological systems mediating this hypothetical construct.     

Shivering and ptiloerection as complementary cold defense responses in the pigeon during sleep and wakefulness

Summary The effects of sleep and wakefulness on two autonomic cold defense responses, shivering and ptiloerection, were studied in the pigeon. Shivering (integrated pectoral muscle EMG), axillary and foot temperatures were measured continuously and the magnitude of ptiloerection was estimated by a specially devised feather index (FI) using a TV-monitor. Behavioral signs of sleep and wakefulness were verified by EEG-recordings.In pigeons placed at +4°C, slow-wave sleep was associated with a gradual decline of shivering and a simultaneous increase of FI. Spontaneous bouts of arousal elicited strong bursts of shivering and coincident drops in FI (Fig. 2). Alerting the sleeping pigeons by a standardized stimulus had the same effects as spontaneous arousal, but the changes were more consistent. The stimulus produced behavioral and electrocortical arousal, a 81% increase in shivering intensity, and an almost complete elimination of ptiloerection. A slight vasodilatation of feet and a neglible increase of axillary temperature followed (Fig. 3). The changes in shivering and FI showed a significant negative correlation both during spontaneous and induced bouts of arousal.These results show that ptiloerection is preferentially used for cold defense during sleep, whereas shivering is recruited during periods of arousal. This complementarity of insulative and thermogenic mechanisms may form a physiological basis for explaining some diurnal patterns in avian bioenergetics.Abbreviations EEG electroencephalogram - EMG electromyogram - FI feather index     

Neuropeptide S: a new player in the modulation of arousal and anxiety

Neuropeptide S (NPS) is a newly identified transmitter that modulates arousal and fear responses. NPS activates an orphan G protein-coupled receptor that is expressed throughout the central nervous system, including brain centers that regulate sleep/wakefulness and anxiety. In contrast, the NPS precursor mRNA is found only in a few discrete nuclei in the brainstem as well as in a few scattered cells in the hypothalamus and amygdala. The most prominent expression of NPS precursor is found in a previously uncharacterized cluster of neurons in the pontine area, located between the noradrenergic locus ceruleus and Barrington's nucleus. Central administration of NPS induces long-lasting arousal and suppresses all stages of sleep. In addition, NPS produces an anxiolytic profile in a variety of behavioral models. The unique pharmacological spectrum of NPS makes it an interesting target for pharmaceutical development. It also enhances our understanding of the neurobiological mechanisms of sleep/wakefulness regulation and the neuronal processing of stress.     

Clinical and physiological aspects of the arousal reaction in young children

This review article addresses current knowledge on the arousal reaction during sleep. Diagnostic criteria of the arousal scoring in young children are provided. The possible protective role of the arousal reaction is discussed, and certain abnormalities of the arousal mechanisms in some patients with different types of sleep apneas are considered. Potential links between an abnormal arousal reaction and the origin of the sudden infant death syndrome are discussed.     

Parent-infant bed-sharing behavior

An evolutionarily informed perspective on parent-infant sleep contact challenges recommendations regarding appropriate parent-infant sleep practices based on large epidemiological studies. In this study regularly bed-sharing parents and infants participated in an in-home video study of bed-sharing behavior. Ten formula-feeding and ten breast-feeding families were filmed for 3 nights (adjustment, dyadic, and triadic nights) for 8 hours per night. For breast-fed infants, mother-infant orientation, sleep position, frequency of feeding, arousal, and synchronous arousal were all consistent with previous sleep-lab studies of mother-infant bed-sharing behavior, but significant differences were found between formula and breast-fed infants. While breast-feeding mothers shared a bed with their infants in a characteristic manner that provided several safety benefits, formula-feeding mothers shared a bed in a more variable manner with consequences for infant safety. Paternal bed-sharing behavior introduced further variability. Epidemiological case-control studies examining bed-sharing risks and benefits do not normally control for behavioral variables that an evolutionary viewpoint would deem crucial. This study demonstrates how parental behavior affects the bed-sharing experience and indicates that cases and controls in epidemiological studies should be matched for behavioral, as well as sociodemographic, variables.     

Thalamocortical dynamics of sleep: roles of purinergic neuromodulation

Thalamocortical dynamics, the millisecond to second changes in activity of thalamocortical circuits, are central to perception, action and cognition. Generated by local circuitry and sculpted by neuromodulatory systems, these dynamics reflect the expression of vigilance states. In sleep, thalamocortical dynamics are thought to mediate "offline" functions including memory consolidation and synaptic scaling. Here, I discuss thalamocortical sleep dynamics and their modulation by the ascending arousal system and locally released neurochemicals. I focus on modulation of these dynamics by electrically silent astrocytes, highlighting the role of purinergic signaling in this glial form of communication. Astrocytes modulate cortical slow oscillations, sleep behavior, and sleep-dependent cognitive function. The discovery that astrocytes can modulate sleep dynamics and sleep-related behaviors suggests a new way of thinking about the brain, in which integrated circuits of neurons and glia control information processing and behavioral output.     

Relationship between the incremental and behavioral responses evoked by stimulation of the ventral anterior nucleus of the rat thalamus]

Low frequency stimulation of nucleus Ventralis Anterior thalami in unanesthetized rats elicits incremental responses and produces, in some conditions, characteristic behavioral reaction. Stimulation thresholds giving rise to behavioral reaction are higher than those allowing to find electrocortical responses. The analysis of their evolution discloses changes which are bound to both of the following parameters: the stimulation frequency and the arousal. The lowest behavioral thresholds are got at 8-10 c/s during relaxed wakefulness whereas the lowest electrocortical thresholds are found at 5-8 c/s during slow-wave sleep.     

State-dependent bidirectional modification of somatic inhibition in neocortical pyramidal cells

Cortical pyramidal neurons alter their responses to input signals depending on behavioral state. We investigated whether changes in somatic inhibition contribute to these alterations. In layer 5 pyramidal neurons of rat visual cortex, repetitive firing from a depolarized membrane potential, which typically occurs during arousal, produced long-lasting depression of somatic inhibition. In contrast, slow membrane oscillations with firing in the depolarized phase, which typically occurs during slow-wave sleep, produced long-lasting potentiation. The depression is mediated by L-type Ca2+ channels and GABA(A) receptor endocytosis, whereas potentiation is mediated by R-type Ca2+ channels and receptor exocytosis. It is likely that the direction of modification is mainly dependent on the ratio of R- and L-type Ca2+ channel activation. Furthermore, somatic inhibition was stronger in slices prepared from rats during slow-wave sleep than arousal. This bidirectional modification of somatic inhibition may alter pyramidal neuron responsiveness in accordance with behavioral state.     

Catecholamines and the sleep-wake cycle I. EEG and behavioral arousal

The exact role of catecholamines (CA) on the sleep-wake cycle is still controversial. Critical analysis of lesion studies tends to suggest a neuromodulatory role for both dopamine (DA) and norepinephrine (NE) on EEG and behavioral arousal. Support for this view is provided by pharmacological studies in which catecholaminergic systems are activated or inhibited. Taken together they show that disturbances in the dynamic balance between neurochemical systems may alter the conditions for wake-triggering mechanisms to express at optimal levels. Large electrolytic or neurotoxic lesions which affect noradrenergic and dopaminergic structures are associated with marked and prolonged EEG changes and decreased behavioral arousal, respectively. In contrast, specific and circumscribed damage restricted to these systems is followed by a transient decrease in waking activity. Thus, results observed after large central lesions are most probably related to destruction of non-catecholaminergic neurons. Inhibition of brain CA synthesis causes behavioral sedation and a decrease in waking activity. Selective pharmacological stimulation of presynaptic alpha-adrenergic (∝ 2) receptors tends to decrease waking, while opposite effects result from ∝2-receptor blockade. Drugs with agonistic activity at postsynaptic alpha-adrenergic (∝1) sites increase EEG desynchronization, but specific blockade of ∝1-receptor does not result in marked decreases of waking EEG. In contrast, treatments which simultaneously block NE and DA receptors significantly affect waking. Beta-adrenergic receptor blockers show no conclusive effects on waking or sleep. Selective DA-receptor agonists induce biphasic effects, with low doses decreasing and large doses increasing cortical desynchronization and motility. Opposite effects are observed in laboratory animals after injection of specific DA-receptor blockers.     

Baroreflexes of the rat. III. Open-loop gain and electroencephalographic arousal

In early studies of humans, baroreflex sensitivity was found to be higher during sleep; however, subsequent observations in several species, including humans, have been at variance with the original reports. Sleep and arousal are behavioral states, and it is difficult to accurately and repeatedly measure baroreflex sensitivity in behaving animals. However, pharmacologically immobilized (neuromuscularly blocked) rats have apparently normal sleep-wakefulness cycles, and baroreflex gain can be measured directly in this preparation. Using the delta band of the EEG (EEG(delta)) as an index of sleep and arousal and open-loop aortic depressor nerve (ADN) stimulation as a baroreflex input, we found that blood pressure (BP) level depended on arousal (r = -0.416; P < 0.0001), and BP baroreflex gain depended on BP level (r = 0.496; P < 0.0001), but that BP baroreflex gain was independent of arousal (r = 0.001; NS). Heart period (HP) was different; although HP level depended on arousal (r = 0.352; P < 0.0001), HP baroreflex gain did not depend on HP level (r = 0.029; NS), and HP baroreflex gain increased with arousal (r = 0.315; P < 0.0001). A partial-correlations analysis showed that the presence of the relationship between BP level and BP baroreflex gain probably attenuated the relationship between arousal and BP gain. The results are consistent 1) with physiological findings showing that arousal attenuates afferent transmission through the nucleus of the solitary tract and enhances sympathoinhibition at the rostral ventrolateral medulla; and 2) with observations in humans and animals showing increased cardiac baroreflex sensitivity during sleep, but little if any effect of sleep on BP baroreflex sensitivity. The findings are relevant to all methods of baroreflex gain estimation that use HP as the index of baroreflex activation.     

Circadian clock resetting by arousal in Syrian hamsters: the role of stress and activity

Circadian rhythms in the Syrian hamster can be markedly phase shifted by 3 h of wheel running or arousal stimulation during their usual daily rest period ("subjective day"). Continuous wheel running is predictive but not necessary for phase shifts of this "nonphotic" type; hamsters aroused by gentle handling without running can also show maximal shifts. By contrast, physical restraint, a standard stress procedure and thus presumably arousing, is ineffective. To resolve this apparent paradox, phase-shifting effects of 3-h sessions of restraint or other stress procedures were assessed. In a preliminary study, phase shifts to arousal by gentle handling were significantly potentiated by the cortisol synthesis inhibitor metyrapone, suggesting that stress-related cortisol release may inhibit phase shifts to arousal. Next, it was confirmed that restraint in the subjective day does not induce phase shifts, but behavioral observations revealed that it also does not sustain arousal. Restraint combined with noxious compressed air blasts did sustain arousal and induced a significant cortisol response compared with arousal by gentle handling but did not induce shifts. Restraint combined with continuous horizontal rotation was also ineffective, as was EEG-validated arousal via confinement to a pedestal over water. However, 3 h of resident-intruder interactions (an intense psychosocial stress) or exposure to an open field (a mild stress) did induce large shifts that were positively correlated with indexes of forward locomotion. The results indicate that large phase shifts associated with arousal in the usual sleep period are neither induced nor prevented by stress per se, but are dependent on the expression of at least low levels of locomotor activity. Sustained arousal alone is not sufficient.     

Relaxin-3 null mutation mice display a circadian hypoactivity phenotype

Characterizing the neurocircuits and neurotransmitters that underlie arousal and circadian sleep/wake patterns is an important goal of neuroscience research, with potential implications for understanding human mental illnesses, such as major depression. Recent anatomical and functional studies suggest that relaxin-3 neurons and their ascending projections contribute to these functions via actions on key cortical, limbic and hypothalamic circuits. This study reports the behavioral phenotype of C57BL/6J backcrossed relaxin-3 knockout (KO) mice. Cohorts of adult, male and female relaxin-3 KO and wild-type (WT) littermate mice were subjected to a battery of behavioral tests to assess sensorimotor function and complex behavior. No overt deficits were detected in motor-coordination, spatial memory, sensorimotor gating, anxiety-like behavior or locomotor behavior in novel environments; and no marked genotype differences were observed in response to a chronic stress protocol. Notably however, compared to WT mice, relaxin-3 KO mice displayed robust hypoactivity during the dark/active phase when provided with free home-cage access to voluntary running wheels. This circadian hypoactivity was reflected by reduced time spent and distance traveled on running wheels, coupled with an increase in the time spent immobile, possibly reflecting increased sleeping. Overall, these studies support a role for relaxin-3 signaling in the control of arousal and sleep/wakefulness, and identify the relaxin-3 KO mouse as a useful model to study this role further.     

Arousal and cardiopulmonary responses to hyperoxic hypercapnia in lambs

Experiments were done to investigate the arousal and cardiopulmonary responses to hyperoxic hypercapnia in 8 lambs. Each lamb was anaesthetized and instrumented for recordings of electrocorticogram, electro-oculogram, nuchal and diaphragm electromyograms and measurements of arterial blood pressure and haemoglobin oxygen saturation. No sooner than 3 days after surgery, measurements were made in quiet sleep and active sleep during control periods when the animal was breathing 21% oxygen and during experimental periods of hyperoxic hypercapnia when the animal was breathing 10% carbon dioxide and 30% oxygen. Hyperoxic hypercapnia was terminated during each epoch by returning the inspired gas mixture to 21% oxygen once the animal aroused from sleep. Arousal occurred from both sleep states during hyperoxic hypercapnia but was delayed in active sleep compared to quiet sleep (active sleep 58 +/- 17 s; quiet sleep 21 +/- 10 s; mean +/- 1SD). There were no significant changes in heart rate or blood pressure during hyperoxic hypercapnia before arousal. However, respiratory rate and diaphragm electrical activity did increase during hyperoxic hypercapnia before arousal. Thus, our data provide evidence that hypercapnia can initiate arousal from sleep in young lambs. The mechanisms responsible for this response are yet to be determined.     

Sleep propensity is modulated by circadian and behavior-induced changes in cutaneous temperature

A close relation between sleep and body temperature has been noted already for a long time. Although a correlation is indisputable, there is at present hardly evidence for a causal involvement of sleep in changes in body temperature. Concerning the reverse, a causal involvement of body temperature in sleep has been demonstrated: if core or skin temperature changes activate thermoregulatory processes aimed at heat loss or heat preservation, sleep is usually disrupted. We have recently proposed that sleep propensity is also affected by more subtle changes in skin temperature, within the thermoneutral range (Van Someren (2000). Chronobiol. Int. 17, 313–354). These changes are likely to modulate the firing properties of thermosensitive neurons in brain areas involved in sleep regulation. Subtle changes in skin temperature occur daily under control of the circadian timing system. They could provide this system with an additional signal pathway to support its neuronal and neurohormonal signals to enforce circadian modulation of sleep propensity. Subtle changes in skin temperature also result from behavior, and could contribute to the changes in sleep propensity resulting from these behaviors. The present review summarizes the neurobiological background and correlational physiological and behavioral data in support of the involvement of skin temperature in the modulation of sleep propensity. It moreover points out the type of experimental investigations needed to support or refute the hypothesis.     

Ecstasy and sleep disturbance: Progress towards elucidating a role for the circadian system

MDMA (ecstasy) is an illicit drug which has pharmacological actions on the serotonin system, leading to a number of physiological and behavioral changes. Research conducted in both animals and humans has focused on how ecstasy use affects systems or functions in which serotonin has a regulatory role including mood, sleep and circadian rhythms. In this paper we review the evidence with respect to changes in sleep and circadian rhythms following ecstasy use. Studies of the subjective measurement of sleep have suggested that there are changes in sleep quality and duration following ecstasy use, while research utilizing objective measures including polysomnog-raphy has highlighted changes in sleep architecture following ecstasy use. Collectively these findings suggest that there are consequences associated with ecstasy use, and the implications of these findings for ecstasy users will be examined. Finally, preliminary evidence from the animal literature implicating ecstasy as having specific effects on the circadian system will be reviewed. A discussion of the limitations of the current evidence for such a claim is presented, and possible directions for future research are explored.

     

Intermittent hypoxia induces transient arousal delay in newborn mice.

Previous studies suggested that defective arousal might be a major mechanism in sleep-disordered breathing such as sudden infant death syndrome and obstructive sleep apnea. In this study, we examined the effects of intermittent hypoxia (IH) on the arousal response to hypoxia in 4-day-old mice. We hypothesized that IH would increase arousal latency, as previously reported in other species, and we measured the concomitant changes in ventilation to shed light on the relationship between breathing and arousal. Arousal was scored according to behavioral criteria. Breathing variables were measured noninvasively by use of whole-body flow plethysmography. In the hypoxic group (n = 14), the pups were exposed to 5% O(2) in N(2) for 3 min and returned to air for 6 min. This test was repeated eight times. The normoxic mice (n = 14) were constantly exposed to normoxia. The hypoxic mice showed a 60% increase in arousal latency (P < 0.0001). Normoxic controls showed virtually no arousals. IH depressed normoxic ventilation below baseline prehypoxic levels, while preserving the ventilatory response to hypoxia. The breathing pattern and arousal responses recovered fully after 2 h of normoxia. We conclude that IH rapidly and reversibly depressed breathing and delayed arousal in newborn mice. Both effects may be due to hypoxia-induced release of inhibitory neurotransmitters acting concomitantly on both functions.