Bernard M. Bergmann

Sleep deprivation in the rat by the disk-over-water method

Chronic sleep deprivation may be required to reveal the most serious physiological consequences of sleep loss, but it usually requires strong stimulation which can obscure the interpretation of effects. The disk-over-water method permits chronic sleep deprivation of rats with gentle physical stimulation that can be equally applied to yoked control rats. A series of studies with this method has revealed little or no pathology in the control rats. The deprived rats show a reliable syndrome that includes temperature changes (which vary with the sleep stages that are lost); heat seeking behavior; increased food intake; weight loss; increased metabolic rate; increased plasma norepinephrine; decreased plasma thyroxine; an increased triiodothyronine-thyroxine ratio; and an increase of an enzyme which mediates thermogenesis by brown adipose tissue. The temperature changes are attributable to excessive heat loss and an elevated thermoregulatory setpoint, both of which increase thermoregulatory load, and the other changes are interpretable as responses to this increased load. This pattern indicates that sleep serves a thermoregulatory function in the rat. The sleep deprived rats also show stereotypic ulcerative and hyperkeratotic lesions localized to the tail and plantar surfaces of the paws, and they die within a matter of weeks; the mediation of these changes is unresolved.

Sleep Deprivation in the Rat: IV. Paradoxical Sleep Deprivation

Twelve rats were subjected to paradoxical sleep deprivation (PSD) by the disk apparatus. All PSD rats died or were sacrificed when death seemed imminent within 16-54 days. No anatomical cause of death was identified. All PSD rats showed a debilitated appearance, lesions on their tails and paws, and weight loss in spite of increased food intake. Their yoked control (PSC) rats remained healthy. Since dehydration was ruled out and several measures indicated normal or accelerated use of nutrients, the food-weight changes in PSD rats were attributed to increased energy expenditure (EE). The measurement of EE, based upon caloric value of food, weight, and wastes, indicated that all PSD rats increased EE, with mean levels reaching more than twice baseline values. All of these changes had been observed in rats deprived totally of sleep; the major difference was that they developed more slowly in PSD rats.

Recovery from Sleep Deprivation Occurs during Propofol Anesthesia

Background: Some neurophysiologic similarities between sleep and anesthesia suggest that an anesthetized state may reverse effects of sleep deprivation. The effect of anesthesia on sleep homeostasis, however, is unknown. To test the hypothesis that recovery from sleep deprivation occurs during anesthesia, the authors followed 24 h of sleep deprivation in the rat with a 6-h period of either ad libitum sleep or propofol anesthesia, and compared subsequent sleep characteristics. Methods: With animal care committee approval, electroencephalographic/electromyographic electrodes and intrajugular cannulae were implanted in 32 rats. After a 7-day recovery and 24-h baseline electroencephalographic/electromyographic recording period, rats were sleep deprived for 24 h by the disk-over-water method. Rats then underwent 6 h of either propofol anesthesia (n = 16) or ad libitum sleep with intralipid administration (n = 16), followed by electroencephalographic/electromyographic monitoring for 72 h. Results: In control rats, increases above baseline in non–rapid eye movement sleep, rapid eye movement sleep, and non–rapid eye movement delta power persisted for 12 h after 24 h of sleep deprivation. Recovery from sleep deprivation in anesthetized rats was similar in timing to that of controls. No delayed rebound effects were observed in either group for 72 h after deprivation. Conclusion: These data show that a recovery process similar to that occurring during naturally occurring sleep also takes place during anesthesia and suggest that sleep and anesthesia share common regulatory mechanisms. Such interactions between sleep and anesthesia may allow anesthesiologists to better understand a potentially important source of variability in anesthetic action and raise the possibility that anesthetics may facilitate sleep in environments where sleep deprivation is common.

Variations in slow wave activity during sleep in the rat

Scoring of human electroencephalogram (EEG) recordings usually includes subdivisions of non-rapid eye movement (NREM) sleep based on amount of slow wave activity. This procedure has revealed relationships between slow wave activity and many other variables. In animals, however, few experimenters have described variations in slow wave activity within NREM sleep. The present study quantifies, by filtering and integration techniques, variations in amount of slow wave activity during NREM sleep in the rat. Slow wave activity was found to be greatest at the start of the light period; the diurnal variation of slow wave activity within NREM sleep was correlated with variations in amount of NREM sleep. An amplitude criterion was used to define NREM sleep, but overall EEG amplitude during NREM sleep did not show the same diurnal variation as slow wave activity. The results indicate the value of measuring variations in slow wave amplitude during NREM sleep in animals in addition to overall EEG amplitude.

Increases in amino-cupric-silver staining of the supraoptic nucleus after sleep deprivation

Sleep deprived rats undergo a predictable sequence of physiological changes, including changes in skin condition, increased energy expenditure, and altered thermoregulation. Amino-cupric-silver staining was used to identify sleep deprivation related changes in the brain. A significant increase in staining was observed in the supraoptic nucleus (SON) of the hypothalamus of rats with high sleep loss (>45 h) vs. their yoked controls. Follow-up experiments showed that staining was not significantly different in rats sleep deprived for less than 45 h, suggesting that injurious sleep deprivation-related processes occur above a threshold quantity of sleep loss. These anatomical changes suggest that the effects of sleep deprivation may be related to protein metabolism in certain brain regions.

Effects of chronic total sleep deprivation on central noradrenergic receptors in rat brain

The effect of chronic total sleep deprivation (TSD) on the regulation of central noradrenergic receptors was evaluated. Rats were subjected to 10 days of TSD by the disk-over-water method. As in previous TSD studies, these rats showed greater increases in food intake and energy expenditure and greater eventual declines in waking body temperature than their yoked-control (TSC) rats. After sacrifice, alpha 1-, alpha 2-, and beta-adrenoceptors were determined in 11 brain regions using radioligand binding assays with [3H]prazosin, [3H]rauwolscine, and 125I-iodocyanopindolol, respectively. Adrenoceptor density and affinity values were significantly different among TSD, TSC, and normally caged control rat groups only for the cerebellum, which showed higher alpha 2-binding density concomitant with lower affinity and lower beta-binding density than cage control rats. Such changes are attributable to apparatus or stimulus effects common to TSD and TSC rats. Given the absence of firm evidence for a TSD-induced downregulation of central noradrenergic receptors, the present results offer no support for the hypothesis of Siegel and Rogawski that a major function of paradoxical sleep is to upregulate these receptors.

Increased Basal REM Sleep But No Difference in Dark Induction or Light Suppression of REM Sleep in Flinders Rats with Cholinergic Supersensitivity

Increased cholinergic sensitivity in the central nervous system has been postulated to account for some of the neuroendocrine abnormalities and sleep disturbances seen in human depressives. The Flinders Sensitive Line (FSL) rats, which exhibit increased sensitivity to cholinergic agents, have been shown to have REM sleep patterns similar to those seen in depressives, including shorter REM sleep latency and increased daily percentage of REM sleep.We studied the response of FSL and control rats to brief dark pulses administered during the normal light period (which are known to stimulate REM sleep in albino rats) and to brief light pulses during the normal dark period (which suppress REM sleep in albino rats) to determine whether these responses are affected by central cholinergic hypersensitivity. FSL rats showed REM sleep patterns indistinguishable from controls during light or dark pulses, which does not support the primary involvement of cholinergic systems in this mechanism of REM sleep regulation.We also examined REM and non-REM (NREM) sleep patterns in FSL rats and their controls to determine whether they show sleep continuity disturbances or decreased sleep intensity as seen in depression. In agreement with an earlier study, we found that FSL rats had more daily REM sleep and accumulated less NREM sleep between REM bouts than controls. Duration of NREM sleep bouts, total daily NREM sleep time, and EEG amplitude of NREM sleep did not differ between FSL and control rats, suggesting that the cholinergic abnormalities in FSL rats do not produce substantial NREM sleep changes.

Age-dependent changes in recovery sleep after 48 hours of sleep deprivation in rats

To characterize possible changes in homeostatic regulation of sleep with aging, we have examined sleep stages during recovery sleep after 48 h of sleep deprivation in young (3 months), middle aged (12 months), and old (24 months) rats. It was found that young and middle aged, in contrast to old rats, had large (21-24%) increases in total sleep time during recovery sleep; the old rats experienced a quantitatively small (8%) but significant rise in total sleep. NREM sleep increased significantly during the recovery period in young and middle aged, but not older rats. High voltage NREM sleep (HS2) declined by 30% during recovery in the young animals, but remained unchanged compared to baseline in the middle aged and old animals. The young and middle aged rats had increases in REM sleep during recovery compared to their baseline by 96% and 93%, respectively, which was significantly greater than a 65% increase during recovery in the old rats. Increases in total sleep and REM sleep during recovery were largely confined to the first 6 h in young and middle aged rats, but maxima for the old rats occurred in the second 6 h.

Rat strain differences in response to dark pulse triggering of paradoxical sleep

Previous studies of inbred rats have shown that Brown Norway (BN) rats had more daily paradoxical sleep (PS) than Lewis (L) rats, while F1 progeny had intermediate amounts, suggesting codominant or polygenic transmission. Amount of PS and the induction of PS episodes may be under separate genetic control. Earlier work had shown that five-minute exposures to cage lights-off every half-hour can trigger PS in outbred albino strains. To explore the genetic controls for PS induction, PS triggering by dark pulse stimulation was examined in L and BN rats. L rats showed a five-fold increase in PS during dark pulse stimulation. Although, as in the earlier study, BN rats had more total daily PS than L rats, they exhibited no dark pulse triggering of PS. Thus L and BN rats show significant strain differences in two independent parameters of PS, and may be a useful model for studying genetic and neurologic factors which regulate PS.

EEG delta power during sleep in young and old rats

Delta EEG power density, which has been viewed as a measure of intensity of NREM sleep, declines across the lifetime in humans, cats, and hamsters, but data in rats have been unclear. It is also uncertain whether older rats differ from younger animals in the degree of change in delta power during recovery sleep following short-term sleep deprivation. We have examined delta power density in NREM sleep under baseline conditions and following 48 h of sleep deprivation in young (3 months), middle-aged (12 months), and older (24 months) rats. The presence or absence of age effects was highly dependent on the method of normalizing the data. When expressed as a fraction of total NREM EEG power, there was no age effect on baseline delta power density, or on the change from baseline to recovery conditions. When expressed as a multiple of delta power in REM under the same condition, the younger rats had higher delta power density than the middle-aged and older rats. For all the ages combined, there was an increase in delta power density in the recovery condition. When examined by age, the younger rats (which started from a higher level of delta power density than the other groups) did not have an increase in delta during recovery; the middle-aged rats tended to, and the older rats (which started from lower baseline levels) significantly increased delta power density in the recovery condition. This suggests that the lower delta power seen during baseline in older rats is not due to decreased ability to generate delta activity.