Cerebrospinal Fluid Hypocretin and Nightmares in Dementia Syndromes.

BACKGROUND/AIMS: Hypocretin promotes wakefulness and modulates REM sleep. Alterations in the hypocretin system are increasingly implicated in dementia. We evaluated relationships among hypocretin, dementia biomarkers, and sleep symptoms in elderly participants, most of whom had dementia. METHODS: One-hundred twenty-six adults (mean age 66.2 ± 8.4 years) were recruited from the Emory Cognitive Clinic. Diagnoses were Alzheimer disease (AD; n = 60), frontotemporal dementia (FTD; n = 21), and dementia with Lewy bodies (DLB; n = 20). We also included cognitively normal controls (n = 25). Participants and/or caregivers completed sleep questionnaires and lumbar puncture was performed for cerebrospinal fluid (CSF) assessments. RESULTS: Except for sleepiness (worst in DLB) and nocturia (worse in DLB and FTD) sleep symptoms did not differ by diagnosis. CSF hypocretin concentrations were available for 87 participants and normal in 70, intermediate in 16, and low in 1. Hypocretin levels did not differ by diagnosis. Hypocretin levels correlated with CSF total τ levels only in men (r = 0.34; p = 0.02). Lower hypocretin levels were related to frequency of nightmares (203.9 ± 29.8 pg/mL in those with frequent nightmares vs. 240.4 ± 46.1 pg/mL in those without; p = 0.05) and vivid dreams (209.1 ± 28.3 vs. 239.5 ± 47.8 pg/mL; p = 0.014). Cholinesterase inhibitor use was not associated with nightmares or vivid dreaming. CONCLUSION: Hypocretin levels did not distinguish between dementia syndromes. Disturbing dreams in dementia patients may be related to lower hypocretin concentrations in CSF.

Sleep/Wake Behavior and EEG Signatures of the TgF344-AD Rat Model at the Prodromal Stage.

Transgenic modification of the two most common genes (APPsw, PS1ΔE9) related to familial Alzheimer's disease (AD) in rats has produced a rodent model that develops pathognomonic signs of AD without genetic tau-protein modification. We used 17-month-old AD rats (n = 8) and age-matched controls (AC, n = 7) to evaluate differences in sleep behavior and EEG features during wakefulness (WAKE), non-rapid eye movement sleep (NREM), and rapid eye movement sleep (REM) over 24-h EEG recording (12:12h dark-light cycle). We discovered that AD rats had more sleep-wake transitions and an increased probability of shorter REM and NREM bouts. AD rats also expressed a more uniform distribution of the relative spectral power. Through analysis of information content in the EEG using entropy of difference, AD animals demonstrated less EEG information during WAKE, but more information during NREM. This seems to indicate a limited range of changes in EEG activity that could be caused by an AD-induced change in inhibitory network function as reflected by increased GABAAR-ß2 expression but no increase in GAD-67 in AD animals. In conclusion, this transgenic rat model of Alzheimer's disease demonstrates less obvious EEG features of WAKE during wakefulness and less canonical features of sleep during sleep.

Postnatal hypoxia evokes persistent changes within the male rat's dopaminergic system.

PURPOSE: Hypoxic insults occurring during the perinatal period remain the leading cause of permanent brain impairment. Severe cognitive and motor dysfunction, as seen in cerebral palsy, will occur in 4-10% of post-hypoxic newborns. Subtle cognitive impairment, apparent in disorders of minimal brain dysfunction will occur in > 3 million post-hypoxic newborns. Analyses of post-hypoxic rodent brains reveal reduced extracellular levels of dopamine, a key neurotransmitter of vigilance, execute function, and behavior. The purpose of this study was to assess whether synaptic levels of dopamine could be enhanced in post-hypoxic, hypodopaminergic rats. METHODS: Newborn male rats were exposed to subtle, repetitive hypoxic insults for 4-6 h per day, during postnatal days 7-11. During adolescence, we quantified dopamine content within the caudate nuclei. We then determined whether extracellular dopamine levels could be increased by injecting the psychostimulant d-amphetamine. We next assessed whether the post-hypoxic rat's response to d-amphetamine would differentially impact place preference behavior when compared with littermate controls. RESULTS: Total tissue content of dopamine was significantly higher in post-hypoxic rats. Injection of d-amphetamine liberated that dopamine which subsequently enhanced extracellular levels. Post-hypoxic rats acquired conditioned place preference for d-amphetamine during the training days. During the testing day, total time spent in the amphetamine-pairing box did not differ between post-hypoxic and control littermates. CONCLUSION: Postnatally occurring hypoxic insults promote remodeling of the dopaminergic system resulting in increased intracellular sequestering of this monoamine. That sequestered dopamine can be released using the psychostimulant d-amphetamine, which did not promote a conditioned place preference any greater than was observed in non-hypoxic littermate controls.

Hypocretin measurement: shelf age of radioimmunoassay kit, but not freezer time, influences assay variability.

The hypothalamic peptide hypocretin 1 (orexin A) may be assayed in cerebrospinal fluid to diagnose narcolepsy type 1. This testing is not commercially available, and factors contributing to assay variability have not previously been comprehensively explored. In the present study, cerebrospinal fluid hypocretin concentrations were determined in duplicate in 155 patient samples, across a range of sleep disorders. Intra-assay variability of these measures was analyzed. Inter-assay correlation between samples tested at Emory and at Stanford was high (r = 0.79, p < 0.0001). Intra-assay correlation between samples tested in duplicate in our center was also high (r = 0.88, p < 0.0001); intra-assay variability, expressed as the difference between values as a percentage of the higher value, was low at 9.4% (SD = 7.9%). Although both time the sample spent in the freezer (r = 0.16, p = 0.04) and age of the kit used for assay (t = 3.64, p = 0.0004) were significant predictors of intra-kit variability in univariate analyses, only age of kit was significant in multivariate linear regression (F = 4.93, p = 0.03). Age of radioimmunoassay kit affects intra-kit variability of measured hypocretin values, such that kits closer to expiration exhibit significantly more variability.

Maternal dietary supplementation with omega-3 polyunsaturated fatty acids confers neuroprotection to the newborn against hypoxia-induced dopamine dysfunction.

INTRODUCTION: Up to 84% of prematurely born infants suffer hypoxic, anoxic, and ischemic insults. Those infants with subsequent behavioral, motor or cognitive dysfunction represent 8-11% of all live births. Yet, no interventions employed during pregnancy attenuate risk of morbidity in those at-risk infants. Dietary supplementation with omega-3 polyunsaturated fatty acids (ω-3 PUFAs) has been shown to reduce stroke-induced neuropathology in rat models emulating this adverse clinical event. To extend those studies we sought to determine whether maternal dietary supplementation with ω-3 PUFAs would confer neuroprotection against hypoxia-induced neurochemical dysfunction in newborn rat pups exposed to repetitive hypoxic insults. METHODS: We provided pregnant rats with either a ω-3 PUFA enriched diet or else a standard rat chow diet. At postnatal day 7, pups were assigned randomly to either repetitive hypoxic insults or repetitive bursts of room air. On postnatal day 12, pups were sacrificed and brain dopamine levels characterized. RESULTS: Baseline brain dopamine levels did not differ between rat pups born to dams who received ω-3 PUFA enriched versus standard rat chow diets. Rat pups born to dams maintained on normal diets, who were exposed to five days of repetitive hypoxic insults, experienced a 57% reduction in striatal dopamine levels accompanied by significant apoptosis. In contrast, ω-3 PUFA-enriched newborn pups experienced no loss in striatal dopamine levels, and only minimal apoptosis. CONCLUSIONS: Our findings suggest that it may be feasible to confer neuroprotection against hypoxia-induced dopamine dysfunction to newborns likely to experience hypoxic insults. This could significantly improve the outcomes of those 8-11% of newborns who would otherwise experience hypoxia-induced behavioral, motor and cognitive dysfunction.

Effects of γ-Aminobutyric Acid Type A Receptor Modulation by Flumazenil on Emergence from General Anesthesia.

BACKGROUND: Transitions into conscious states are partially mediated by inactivation of sleep networks and activation of arousal networks. Pharmacologic hastening of emergence from general anesthesia has largely focused on activating subcortical monoaminergic networks, with little attention on antagonizing the γ-aminobutyric acid type A receptor (GABAAR). As the GABAAR mediates the clinical effects of many common general anesthetics, the authors hypothesized that negative GABAAR modulators would hasten emergence, possibly via cortical networks involved in sleep. METHODS: The authors investigated the capacity of the benzodiazepine rescue agent, flumazenil, which had been recently shown to promote wakefulness in hypersomnia patients, to alter emergence. Using an in vivo rodent model and an in vitro GABAAR heterologous expression system, they measured flumazenil's effects on behavioral, neurophysiologic, and electrophysiologic correlates of emergence from isoflurane anesthesia. RESULTS: Animals administered intravenous flumazenil (0.4 mg/kg, n = 8) exhibited hastened emergence compared to saline-treated animals (n = 8) at cessation of isoflurane anesthesia. Wake-like electroencephalographic patterns occurred sooner and exhibited more high-frequency electroencephalography power after flumazenil administration (median latency ± median absolute deviation: 290 ± 34 s) compared to saline administration (473 ± 186 s; P = 0.042). Moreover, in flumazenil-treated animals, there was a decreased impact on postanesthesia sleep. In vitro experiments in human embryonic kidney-293T cells demonstrated that flumazenil inhibited isoflurane-mediated GABA current enhancement (n = 34 cells, 88.7 ± 2.42% potentiation at 3 µM). Moreover, flumazenil exhibited weak agonist activity on the GABAAR (n = 10 cells, 10.3 ± 3.96% peak GABA EC20 current at 1 µM). CONCLUSIONS: Flumazenil can modulate emergence from isoflurane anesthesia. The authors highlight the complex role GABAARs play in mediating consciousness and provide mechanistic links between emergence from anesthesia and arousal.

Wake promoting effects of cocaine and amphetamine-regulated transcript (CART).

Cocaine- and amphetamine-regulated transcript (CART) peptides modulate anxiety, food intake, endocrine function, and mesolimbic dopamine related reward and reinforcement. Each of these disparate behaviors takes place during the state of wakefulness. Here, we identify a potential wake promoting role of CART by characterizing its effects upon sleep/wake architecture in rats. Dose-dependent increases in wake were documented following intracerebroventricular CART 55-102 administered at the beginning of the rat's major sleep period. Sustained wake was observed for up to 4h following delivery of 2.0 microg of CART peptide. The wake promoting effect was specific to active CART 55-102 because no effect on sleep/wake was observed with the inactive form of the peptide. Increased wake was followed by robust rebound in NREM and REM sleep that extended well into the subsequent lights-off, or typical wake period, of the rat. These findings point to a potential novel role for CART in regulating wakefulness.

High dose CART peptide induces abnormal EEG activity and behavioral seizures.

Cocaine- and amphetamine-regulated transcript (CART) peptides are neurotransmitters found throughout the nervous system and in the periphery. CART has an important role in the regulation of food intake, anxiety, endocrine function, and in mesolimbic-mediated reward and reinforcement. This short report casts light upon previous descriptions of presumed behavioral seizure and tremor activity following administration of CART into the central nervous system. By employing electroencephalographic (EEG) recordings, we document the state of cerebrocortical activity. We find that intracerebroventricular (icv) administration of 5 microg of CART 55-102 readily produces an abnormal EEG characterized initially by high amplitude hypersynchronous alpha in the 8-10 Hz range during behavioral wakefulness as manifest in both cortical and hippocampal theta EEG channels. This reliably progressed in three of three animals tested to unequivocal epileptiform activity accompanied by tremors and assumption of a rigid, tonic body posture. The neural substrates underlying this finding are unclear. This novel description of the epileptogenic quality of CART should lend caution to interpretations of the behaviors attributed to CART in other experimental paradigms.

The pedunculopontine tegmental nucleus and responding for sucrose reward.

Pedunculopontine tegmental nucleus (PPTg) lesions in rodents lead to increased sucrose consumption, but the psychological deficit behind this remains uncertain. To understand better the relationship between consumption of, and motivation for, sucrose, the authors trained rats to traverse a runway for 20% or 4% sucrose solution; after 7 days, concentrations were reversed. Control rats consumed more 20% than 4% sucrose solution and promptly altered run times in response to concentration change. PPTg-lesioned rats consumed normal quantities of 4% but more 20% sucrose solution than controls and took longer to alter their runway time following the concentration change. These data suggest that lesions of the PPTg do not alter motivation per se and might be better understood as inducing a response selection deficit.

Reduced extracellular dopamine and increased responsiveness to novelty: neurochemical and behavioral sequelae of intermittent hypoxia.

STUDY OBJECTIVES: Vesicular monoamine transporter and dopamine D1-receptor protein expression are upregulated within the striatum of adults rats exposed to intermittent hypoxic insults as neonates. These observations prompted us to test the hypothesis that intermittent hypoxic insults, occurring during this period of critical brain development, lead to persistent reductions in extracellular levels of dopamine within the striatum. We also tested the hypothesis that post-hypoxic rats exhibit increased novelty-induced behavioral activation and increased basal levels of locomotor activity, two indexes of impaired dopaminergic functioning. STUDY DESIGN: Extracellular levels of dopamine were measured over a 50-hour interval via in-vivo microdialysis. Responsivity to novelty and basal levels of locomotor activity were measured via cubicles equipped with infrared transmitters and photosensors. INTERVENTIONS: Between postnatal days 7 and 11, rat pups were exposed to intermittent hypoxia (20-second bursts of hypoxic gas consisting of 10% oxygen balance nitrogen; 60 events/hour) or compressed air for 6 hours during their major sleep period. On postnatal day 35, locomotor activity was continuously recorded over a 72-hour period. On postnatal day 42, extracellular levels of dopamine were determined in microdialysis samples collected at 30-minute intervals continuously for 50 hours. MEASUREMENTS AND RESULTS: Post-hypoxic rats exhibited heightened behavioral activation when placed into the novel environment of locomotor-recording cubicles. Following 24 hours of adaptation to the cubicles, post-hypoxic rats exhibited locomotor hyperactivity during each dark phase of the circadian cycle, their typical waking period. These same rats also demonstrated reduced levels of extracellular dopamine during both the dark and light phases of the circadian cycle. CONCLUSIONS: We demonstrate increased responsivity to novelty, locomotor hyperactivity, and reduced levels of extracellular dopamine within the striata of juvenile rats exposed to intermittent hypoxic insults between postnatal days 7 and 11. These data, in conjunction with our previous observations, support our hypothesis that intermittent hypoxic insults occurring during a period of critical brain development lead to sequestration of dopamine presynaptically within nigrostriatal axons. We postulate that neonatally occurring hypoxic insults are one potential pathogenic mechanism underlying disorders of minimal brain dysfunction, such as attention-deficit/hyperactivity disorder, characterized by executive dysfunction and hyper responsiveness to novel stimuli, which is responsive to agents promoting enhanced extracellular levels of synaptic dopamine.

An examination of the effects of bilateral excitotoxic lesions of the pedunculopontine tegmental nucleus on responding to sucrose reward.

The effects of bilateral excitotoxic lesions of the pedunculopontine tegmental nucleus (PPTg) on sucrose intake were examined in three experiments. First, in tests of conditioned place preference using 20% sucrose as the reinforcer, it was shown that lesioned rats, regardless of whether they were food deprived or non-deprived, formed normal place preferences and showed normal amounts of locomotion. However, consumption of 20% sucrose in the pairing trials was increased in the deprived PPTg lesioned rats compared to their matched controls. A second experiment showed that sucrose consumption in the home cage was increased in both deprived and non-deprived PPTg lesioned rats, but only when the concentration of sucrose was greater than 12%: below this there were no differences in intake between the lesioned and control rats. In a third home cage experiment, it was again shown that non-deprived PPTg lesioned rats increased their consumption of 20% sucrose compared to controls. PPTg lesioned rats concomitantly reduced their intake of lab chow such that overall energy intake remained the same as that of control rats. These data are taken to suggest (i) that bilateral excitotoxic lesions of the PPTg increase consumption of sucrose selectively in conditions of high motivational excitement; (ii) that the perception of the rewarding value of 20% sucrose, as judged by place preference, is not affected by these lesions; and (iii) that PPTg lesioned rats are able to adjust their energy intake to accommodate increased sucrose loads. These data are consistent with the hypothesis that bilateral excitotoxic lesions of the PPTg do not affect energy balance regulation or judgment of the hedonic value of sucrose, but that they do affect the control of responding in the face of high levels of motivational excitement.