Giovanni Zamboni

Quantitative analysis of short term deprivation and recovery of desynchronized sleep in cats

A quantitative analysis of desynchronized sleep (DS) deprivation (exposure to low ambient temperature) and recovery was carried out in unrestrained cats. The results show that: (1) the circadian obligate quota of DS is precisely regulated by a control mechanism paying by the 24th h the DS debt induced by up to 14 h of total DS deprivation, if recovery occurs during the rest phase of circadian photoperiodicity (CP); (2) during static rebound DS hourly duration is increased by 40 +/- 4 sec/h of total deprivation (mean and S.E.) and there is a statistically significant increase in the hourly frequency and in the duration of DS episodes, with respect to control values; (3) the hourly obligate and facultative quotas of DS amount to 409 +/- 82 and 229 +/- 140 sec/h (mean and S.D.), respectively. (4) preoptic heating during deprivation at low ambient temperature decreases the DS static rebound during recovery at control ambient temperature by an amount corresponding to the cumulative duration of DS episodes elicited by the preoptic treatment; (5) preoptic temperature decreases the more markedly the shorter the duration of exposure to low ambient temperature and regains control values the sooner the smaller the DS debt incurred during deprivation; (6) DS debt cumulation is a continuous process which develops at a steady rate during day and night, whereas DS debt payment is a discontinuous process (ultradian rhythm of DS) which develops at a variable rate depending on the DS debt and on the phase of CP.

Threshold of respiratory responses to preoptic heating during sleep in freely moving cats.

Abstract The respiratory effects of unilateral preoptic heating were studied during sleep in freely moving cats. Hypothalamic temperature was recorded by means of a thermistor placed about 5 mm posteriorly to the caudal electrode of the heating pair. Environmental temperature was kept at about 24–28 °C to maintain panting threshold low. Preoptic heating, increasing the hypothalamic temperature by 0.047 ± 0.001 °C, elicited thermal polypnoea or panting during slow-wave sleep, whereas during fast-wave sleep it raised hypothalamic temperature by 0.050 ± 0.003 °C above the value of treated slow-wave sleep but did not result in thermal polypnoea or panting. Only stronger heating, applied to the preoptic region until the hypothalamic temperature increase averaged 0.131 ± 0.009 °C above the value of treated slow-wave sleep, elecited thermal polypnoea during fast-wave sleep. The results show that the threshold of respiratory responses to preoptic heating is consistently increased during fast-wave sleep. The implications of this conclusion are discussed in terms of the homeostatic function of the hypothalamus.

Pattern of desynchronized sleep during deprivation and recovery induced in the rat by changes in ambient temperature

SUMMARY  The pattern of desynchronized sleep (DS) occurrence in the rat was studied during exposure to an ambient temperature (Ta) of 0°C for 48 h and during a 12 h recovery period at laboratory Ta (23°C) following the first and second 24 h of cold exposure. The exposure to low Ta induces a DS deprivation which is followed, during recovery, by a clear DS rebound. Both the decrease and the following increase in the amount of DS are due to changes in the frequency rather than in the duration of DS episodes.

The influence of a heavy thermal load on REM sleep in the rat.

This study was carried out in order to further test the hypothesis that the occurrence of REM sleep in the rat in the form of episodes separated by long intervals (single REM sleep episodes) and by short intervals (sequential REM sleep episodes) is differently influenced by changes in both sleep and ambient related processes. Rats were studied during the exposure to Ta -10 degrees C for 24 or 48 h and during a 12 h recovery period at laboratory Ta (23 degrees C) following either the first or the second 24 h of cold exposure. The exposure to such a low Ta induced an almost complete abolition of REM sleep which was followed, during recovery, by a marked REM sleep rebound. However, in spite of the larger REM sleep deprivation, the REM sleep rebound was weaker following the 48 h-exposure than that following the exposure for 24 h. The increase in the amount of REM sleep during the recovery period was due to an increase in the amount of that occurring in the form of sequential episodes, whilst that in the form of single episodes did not change with respect to control levels. However, the occurrence of REM sleep in the form of sequential episodes was partially impaired during the REM sleep rebound observed in the recovery period following the 48 h-exposure. These results would suggest that the homeostatic regulation of physiological variables may conflict with that of REM sleep occurrence and that the degree of such a contrast is indicated, at low Ta, by the amount of REM sleep in the form of single episodes and, during the following recovery, by the amount of REM sleep in the form of sequential episodes.

The Inhibition of Neurons in the Central Nervous Pathways for Thermoregulatory Cold Defense Induces a Suspended Animation State in the Rat

The possibility of inducing a suspended animation state similar to natural torpor would be greatly beneficial in medical science, since it would avoid the adverse consequence of the powerful autonomic activation evoked by external cooling. Previous attempts to systemically inhibit metabolism were successful in mice, but practically ineffective in nonhibernators. Here we show that the selective pharmacological inhibition of key neurons in the central pathways for thermoregulatory cold defense is sufficient to induce a suspended animation state, resembling natural torpor, in a nonhibernator. In rats kept at an ambient temperature of 15°C and under continuous darkness, the prolonged inhibition (6 h) of the rostral ventromedial medulla, a key area of the central nervous pathways for thermoregulatory cold defense, by means of repeated microinjections (100 nl) of the GABAA agonist muscimol (1 mm), induced the following: (1) a massive cutaneous vasodilation; (2) drastic drops in deep brain temperature (reaching a nadir of 22.44 ± 0.74°C), heart rate (from 440 ± 13 to 207 ± 12 bpm), and electroencephalography (EEG) power; (3) a modest decrease in mean arterial pressure; and (4) a progressive shift of the EEG power spectrum toward slow frequencies. After the hypothermic bout, all animals showed a massive increase in NREM sleep Delta power, similarly to that occurring in natural torpor. No behavioral abnormalities were observed in the days following the treatment. Our results strengthen the potential role of the CNS in the induction of hibernation/torpor, since CNS-driven changes in organ physiology have been shown to be sufficient to induce and maintain a suspended animation state.

Cutaneous vasodilation elicited by disinhibition of the caudal portion of the rostral ventromedial medulla of the free-behaving rat.

Putative sympathetic premotor neurons controlling cutaneous vasomotion are contained within the rostral ventromedial medulla (RVMM) between levels corresponding, rostrally, to the rostral portion of the nucleus of the facial nerve (RVMM(fn)) and, caudally, to the rostral pole of the inferior olive (RVMM(io)). Cutaneous vasoconstrictor premotor neurons in the RVMM(fn) play a major role in mediating thermoregulatory changes in cutaneous vasomotion that regulate heat loss. To determine the role of neurons in the RVMM(io) in regulating cutaneous blood flow, we examined the changes in the tail and paw skin temperature of free-behaving rats following chemically-evoked changes in the activity of neurons in the RVMM(io). Microinjection of the GABA(A) agonist, muscimol, within either the RVMM(fn) or the RVMM(io) induced a massive peripheral vasodilation; microinjection of the GABA(A) antagonist bicuculline methiodide within the RVMM(fn) reversed the increase in cutaneous blood flow induced by warm exposure and, unexpectedly, disinhibition of RVMM(io) neurons produced a rapid cutaneous vasodilation. We conclude that the tonically-active neurons driving cutaneous vasoconstriction, likely sympathetic premotor neurons previously described in the RVMM(fn), are also located in the RVMM(io). However, in the RVMM(io), these are accompanied by a population of neurons that receives a tonically-active GABAergic inhibition in the conscious animal and that promotes a cutaneous vasodilation upon relief of this inhibition. Whether the vasodilator neurons located in the RVMM(io) play a role in thermoregulation remains to be determined.

Hypothalamic temperature during desynchronized sleep

SummaryThe influence of changes in heat loss at specific heat exchangers, appraised by recording ear skin temperature (Ts), on the hypothalamic temperature (Thy) rise during desynchronized sleep (DS) was studied in unrestrained cats at different ambient temperatures (Ta). The correlations of initial and final Thy changes (ΔiThy and ΔfThy) with initial and final Ts changes (ΔiTs and ΔfTs), respectively, are statistically not significant. However, a significant negative correlation was found between ΔfiThy, i.e. ΔfThy-ΔiThy, and ΔfiTs, i.e. ΔfTs-ΔiTs. The latter result shows that only after its initial rise is Thy affected by changes in heat loss at specific heat exchangers during DS. Accordingly, ΔfThy and ΔfiThy are positively correlated with Ta. However, also ΔiThy, although not affected by heat loss changes at specific heat exchangers, shows a positive correlation with Ta. On the basis of these results and of the data in the literature, it is concluded that three independent factors underlie Thy changes during DS, namely heat loss at specific heat exchangers, hypothalamic blood flow and thermogenesis.

Changes in REM sleep occurrence due to rhythmical auditory stimulation in the rat

The effects of the rhythmical delivery of an auditory stimulus (1000 Hz, from 50 to 100 dB, 20 ms, every 20 s) on the pattern of rapid eye movement (REM) sleep occurrence was studied in the rat. The stimulation was simultaneously carried out on pairs of rats over 5 consecutive days (10-h recording sessions), during which a tone of increasing intensity (50, 63, 75, 88, 100 dB) was used. In each experimental session, auditory stimulation was triggered by the REM sleep occurrence of one rat (REMS-selective stimulation) whilst the other rat simultaneously received the same stimuli, but during any stage of the wake-sleep cycle (REMS-unselective stimulation). The results showed that the total amount of REM sleep in the 10-h recording session was increased over the 5 days of stimulation in the REMS-unselective group. This effect was due to an increase in the mean duration of REM sleep episodes. However, no significant changes were observed in animals under REMS-selective stimulation, nor in a third group of animals in which the spontaneous evolution of REM sleep occurrence (REMS-spontaneous) was studied. Since 86% of the stimuli under the REMS-unselective auditory stimulation fell outside REM sleep, the result would suggest that REM sleep occurrence is affected when the stimuli are delivered during a time period (i.e. during wakefulness or non-REM sleep) in which it is well known that physiological regulations are fully operant.

Pattern of REM sleep occurrence in continuous darkness following the exposure to low ambient temperature in the rat.

The occurrence of REM sleep episodes, separated by intervals >3 min (single episodes) and < or =3 min (sequential episodes), was determined in the rat during the recovery (ambient temperature (Ta) 23 degrees C, L period of the LD [12 h:12 h]-cycle), which followed the exposure to low Ta (0 and -10 degrees C) during the D period of the previous LD-cycle, either in normal light (DL) or in continuous darkness (DD). Both exposures were characterized by an almost complete disappearance of REM sleep, whilst the recoveries showed an increase in the amount of REM sleep in the form of sequential episodes, which in DD was particularly prominent and concomitant with a decrease in the amount of REM sleep in the form of single episodes. The initial 2 h-rate of REM sleep occurrence was lower following the exposure to Ta -10 degrees C, than to Ta 0 degrees C. In DD, such an effect was due to the large reduction in the occurrence of sequential REM sleep episodes. A functional correlate of this finding is that the accumulation capacity of a second messenger (cAMP) was found to be lower at the end of the exposure to Ta -10 degrees C, with respect to both the control (Ta 23 degrees C) and the end of exposure to Ta 0 degrees C, in the preoptic-anterior hypothalamus, but not in the cerebral cortex.

The short-term effects of dl-propranolol on the wake-sleep cycle of the rat are related to selective changes in preoptic cyclic AMP concentration

SummaryThe short-term effects of the intraperitoneal administration of dl-propranolol on the wake-sleep cycle of the rat were studied in relation to the cyclic AMP concentration in the preoptic region and cerebral cortex. The results show that propranolol, but not saline, affected all stages of the wake-sleep cycle, increasing wakefulness, decreasing synchronized sleep and abolishing desynchronized sleep. These effects were associated with a decrement in cyclic AMP concentration both in wakefulness and synchronized sleep. However, this decrement was relatively larger in the preoptic region than in the cerebral cortex. The effects of the drug on cyclic AMP accumulation were also studied in hypoxia, a condition of unspecific brain stimulation. In this condition, the cyclic AMP concentration in both brain regions was found to be higher than that observed during either wakefulness or synchronized sleep. In the hypoxic condition propranolol was found to decrease the nucleotide concentration to the same levels observed in wakefulness and synchronized sleep following its administration. However, no difference in the relative magnitude of the decrement was found between the preoptic region and the cerebral cortex. These findings suggest that in both brain regions the drug acts on a cyclic AMP accumulating system, which may be defined as propranololsensitive. The activity of the propranolol-sensitive system in the preoptic region would appear to be related to wake-sleep processes.