Blockade of GABA, type A, receptors in the rat pontine reticular formation induces rapid eye movement sleep that is dependent upon the cholinergic system.

The brainstem reticular formation is an area important to the control of rapid eye movement (REM) sleep. The antagonist of GABA-type A (GABA(A)) receptors, bicuculline methiodide (BMI), injected into the rat nucleus pontis oralis (PnO) of the reticular formation resulted in a long-lasting increase in REM sleep. Thus, one factor controlling REM sleep appears to be the number of functional GABA(A) receptors in the PnO. The long-lasting effect produced by BMI may result from secondary influences on other neurotransmitter systems known to have long-lasting effects. To study this question, rats were surgically prepared for chronic sleep recording and additionally implanted with guide cannulas aimed at sites in the PnO. Multiple, 60 nl, unilateral injections were made either singly or in combination. GABA(A) receptor antagonists, BMI and gabazine (GBZ), produced dose-dependent increases in REM sleep with GBZ being approximately 35 times more potent than BMI. GBZ and the cholinergic agonist, carbachol, produced very similar results, both increasing REM sleep for about 8 h, mainly through increased period frequency, with little reduction in REM latency. Pre-injection of the muscarinic antagonist, atropine, completely blocked the REM sleep-increase by GBZ. GABAergic control of REM sleep in the PnO requires the cholinergic system and may be acting through presynaptic modulation of acetylcholine release.

Enhancement of rapid eye movement sleep in the rat by actions at A1 and A2a adenosine receptor subtypes with a differential sensitivity to atropine.

The adenosine agonist cyclohexaladenosine injected into the medial pontine reticular formation of the rat induces a long-lasting increase in rapid eye movement sleep. To investigate the adenosine receptor-subtype(s) mediating this effect, the dose-response relationships for increasing rapid eye movement sleep by two highly selective adenosine receptor agonists were compared. Rats were surgically prepared for chronic sleep recording and bilateral guide cannulae were aimed at medial sites in the caudal, oral pontine reticular formation. Injections were made unilaterally in 60 nl volumes within 1 h after lights-on. The adenosine agonists used were A1-selective cyclohexaladenosine (10(-6)-10(-4) M) and A2a-selective CGS 21680 (10(-7)-10(-3) M). Each animal also received a series of three, paired-consecutive injections of the muscarinic receptor antagonist atropine (4x10(-3) M) followed by the lowest effective dose of each agonist or saline as control. The A2a receptor agonist, CGS 21680, was one order of magnitude more potent than the A1 receptor agonist, cyclohexaladenosine, in inducing rapid eye movement sleep increases. Preinjection of atropine at a dose that did not itself affect rapid eye movement sleep resulted in antagonism of CGS 21680, but not cyclohexaladenosine-induced rapid eye movement sleep. The differential sensitivity of these ligands to antagonism by atropine supports the conclusion that both A1 and A2a adenosine receptor subtypes in the reticular formation subserve agonist-induced rapid eye movement sleep and that they do so by independent mechanisms. The A2a mechanism requires the cholinergic system and may act through the increased release of acetylcholine. The A1 mechanism operates at a different locus possibly through an inhibition of GABA neurotransmission.

Rapid eye movement sleep deprivation modifies expression of long-term potentiation in visual cortex of immature rats.

During rapid eye movement (REM) sleep, activity of non-retinal origin is propagated into central visual-system pathways in a manner similar, in pattern and intensity, to central visual-system activity that is exogenously generated in waking. It has been hypothesized that REM sleep, which is more abundantly represented early in life than later, functions to provide adjunct 'afferent' input for shaping synaptic connectivity during brain maturation. Here we present data that support this proposal. Recent studies have described a developmentally regulated form of in vitro long-term potentiation (LTP) in the visual cortex that is experience- and age-dependent. In immature rats, suppression of retinal activation of the visual system by removal of visual experience (dark rearing) extends the age when the developmentally regulated form of LTP can be produced. This study tests whether suppression of REM-state activation of the visual system also lengthens the developmental period in which this specific form of LTP can be elicited. Young rats were deprived of REM sleep by the multiple-small-platforms-over-water method during the typically latest week for induction of such LTP in slices of visual cortex. After this week, we could still induce LTP in slices from nearly all the REM-sleep-deprived rats (8/9) but not from age-matched rats that had not lost REM sleep (0/5). The control rats had been housed on large platforms that allow the animals to obtain REM sleep. Only body weights and the concentration of thyrotrophin-releasing hormone in the hypothalamus distinguished home-caged, normal-sleeping controls from both groups of platform animals. On all measures, stress levels were not dissimilar in the two platforms groups. After 7 days of behavioral suppression of REM sleep in immature rats, and consequent reduction of the intense, extra-retinal activity endogenously generated during this sleep state, we found that the period was extended in which developmentally regulated synaptic plasticity (LTP) could be elicited in slices of visual neocortex. These studies support the role of REM sleep and its associated neuronal activity in brain maturation.

Infusion of adenylyl cyclase inhibitor SQ22,536 into the medial pontine reticular formation of rats enhances rapid eye movement sleep.

Microinjection of cholinergic and adenosinergic agonists into the medial pontine reticular formation of rats produces long lasting increases in the time spent in rapid eye movement sleep. Several G-protein-coupled muscarinic and adenosinergic receptors share a common action of inhibition of adenylyl cyclase and inhibition of cyclic adenosine monophosphate. Inhibition of cyclic adenosine monophosphate has been implicated in the mechanism of rapid eye movement sleep induction in the cat. We sought to determine whether a direct inhibitor of adenylyl cyclase microinjected into the rat reticular formation at sites where muscarinic and adenosinergic agonists are effective in producing long lasting elevations in rapid eye movement sleep also result in similar effects on the sleep/wake cycle. The caudal, oral pontine reticular formation was unilaterally infused with 60 nl volumes of carbachol (0.1-1.1mM) and N(6)-cyclohexyladenosine (0.1mM) each within 1h of lights on. Sites effective for significantly elevating rapid eye movement sleep for the 8h following microinjection of both receptor agonists were additionally injected with the adenylyl cyclase inhibitor, SQ22,536 (0.1M). Pontine injections of SQ22,536 resulted in significant mean increases in rapid eye movement sleep time and rapid eye movement sleep period frequency at all of these sites. As with the receptor agonists, SQ22,536 did not alter latency to rapid eye movement sleep onset. Rapid eye movement sleep amounts were observed to be significantly elevated by SQ22,536 at two days, but not at four days, following a single microinjection. These data implicate inhibition of cyclic adenosine monophosphate in the pons of the rat as a mechanism involved in the long-term modulation of rapid eye movement sleep. This mechanism may underlie the homeostatic regulation exhibited by this sleep-state.

Increased gamma- and decreased delta-oscillations in a mouse deficient for a potassium channel expressed in fast-spiking interneurons.

Kv3.1 is a voltage-gated, fast activating/deactivating potassium (K(+)) channel with a high-threshold of activation and a large unit conductance. Kv3.1 K(+) channels are expressed in fast-spiking, parvalbumin-containing interneurons in cortex, hippocampus, striatum, the thalamic reticular nucleus (TRN), and in several nuclei of the brain stem. A high density of Kv3.1 channels contributes to short-duration action potentials, fast afterhyperpolarizations, and brief refractory periods enhancing the capability in these neurons for high-frequency firing. Kv3.1 K(+) channel expression in the TRN and cortex also suggests a role in thalamocortical and cortical function. Here we show that fast gamma and slow delta oscillations recorded from the somatomotor cortex are altered in the freely behaving Kv3.1 mutant mouse. Electroencephalographic (EEG) recordings from homozygous Kv3.1(-/-) mice show a three- to fourfold increase in both absolute and relative spectral power in the gamma frequency range (20-60 Hz). In contrast, Kv3.1-deficient mice have a 20-50% reduction of power in the slow delta range (2-3 Hz). The increase in gamma power is most prominent during waking in the 40- to 55-Hz range, whereas the decrease in delta power occurs equally across all states of arousal. Our findings suggest that Kv3. 1-expressing neurons are involved in the generation and maintenance of cortical fast gamma and slow delta oscillations. Hence the Kv3. 1-mutant mouse could serve as a model to study the generation and maintenance of fast gamma and slow delta rhythms and their involvement in behavior and cognition.

Ponto-geniculo-occipital-wave suppression amplifies lateral geniculate nucleus cell-size changes in monocularly deprived kittens.

We have previously shown that during the post-natal critical period of development of the cat visual system, 1 week of instrumental rapid eye movement (REM) sleep deprivation (IRSD) during 2 weeks of monocular deprivation (MD) results in significant amplification of the effects of solely the 2-week MD on cell-size in the binocular segment of the lateral geniculate nucleus (LGN) [36,40]. In this study, we examined whether elimination of ponto-geniculo-occipital (PGO)-wave phasic activity in the LGN during REM sleep (REMS), rather than suppression of all REMS state-related activity, would similarly yield enhanced plasticity effects on cell-size in LGN. PGO-activity was eliminated in LGN by bilateral pontomesencephalic lesions [8,32]. This method of removing phasic activation at the level of the LGN preserved sleep and wake proportions as well as the tonic activities (low voltage, fast frequency ECoG and low amplitude EMG) that characterize REM sleep. The lesions were performed in kittens on post-natal day 42, at the end of the first week of the 2-week period of MD, the same age when IRSD was started in the earlier study. LGN interlaminar cell-size disparity increased in the PGO-wave-suppressed animals as it had in behaviorally REM sleep-deprived animals. Smaller A1/A-interlaminar ratios reflect the increased disparity effect in both the REM sleep- and PGO-suppressed groups compared to animals subjected to MD-alone. With IRSD, the effect was achieved because the occluded eye-related, LGN A1-lamina cells tended to be smaller relative to their size after MD-alone, whereas after PGO-suppressing lesions, the A1-lamina cells retained their size and the non-occluded eye-related, A-lamina cells tended to be larger than after MD-alone. Despite this difference, for which several possible explanations are offered, these A1/A-interlaminar ratio data indicate that in conjunction either with suppression of the whole of the REMS state or selective removal of REM sleep phasic activity at the LGN, altered visual input evokes more LGN cell plasticity during the developmental period than it would otherwise. These data further support involvement of the REM sleep state in reducing susceptibility to plasticity changes and undesirable variability in the course of normative CNS growth and maturation.

Neuronal activity in the lateral geniculate nucleus associated with ponto-geniculo-occipital waves lacks lamina specificity.

Ponto-geniculo-occipital (PGO) waves are spontaneously occurring field potentials recorded in the dorsal lateral geniculate nucleus (LGN) just prior to and during rapid eye movement (REM) sleep. Facilitated discharge rates of LGN neurons are associated with PGO waves. In kittens during the critical period of visual system development, both visual experience and PGO waves appear capable of influencing the course of development through activity-dependent mechanisms. Retinal innervation of LGN segregates into eye-specific laminae and is critical to supporting the role of binocular visual experience in development. We sought to determine whether neuronal activity associated with PGO waves also exhibits lamina specificity. PGO wave-related discharges were examined in LGN neurons identified as to lamina location in adult cats administered urethane anesthesia and the reserpine-like compound, RO4-1284. Spontaneous activity of LGN neurons was related to the occurrence of PGO-like waves in all cells studied. No factors could be found that differentiated lamina location and PGO wave-related discharges. We conclude that the PGO wave influence on neuronal activity in the visual system is fundamentally different from that derived from visual experience. The implications of this difference for the role of the two sources of activation in the control of neural activity in development are discussed.

Enhancement of rapid eye movement sleep in the rat by cholinergic and adenosinergic agonists infused into the pontine reticular formation.

The cholinergic agonist carbachol (1.1 mM) and the adenosinergic agonist cyclohexyladenosine (0.1 mM) were microinjected (60 nl) into the region of the caudal, oral pontine reticular formation of the rat. Local intracerebral infusion of each receptor agonist resulted in significant, long-lasting (at least 8 h) elevations in rapid eye movement sleep without reduction in latency to onset. The effects of carbachol were reduced by the muscarinic receptor antagonist atropine, while those of cyclohexyladenosine were reduced by the adenosinergic receptor antagonist 8-cyclopentyltheophylline. Atropine failed to antagonize the long-term induction of rapid eye movement sleep following cyclohexyladenosine, but did appear to suppress increases in the first 2 h. Similarity of effects on sleep parameters and the lack of additivity when injected consecutively are consistent with these agonist ligands targeting the same cellular mechanisms through their respective receptors. These findings suggest that transitory increases in the pons of either acetylcholine or adenosine may underlie long-lasting elevations in the amount of rapid eye movement sleep. Adenosine may play a role in the increased rapid eye movement sleep following prolonged wakefulness, as well as following conditions of stress and learning.

REM sleep deprivation in monocularly occluded kittens reduces the size of cells in LGN monocular segment.

STUDY OBJECTIVES: In this study, we test the hypothesis that when REM-state activation (which impinges upon all lateral geniculate nucleus laminae irrespective of stimulating eye) is deprived, the monocular segment (MS) that is cut off from visual input and also deprived of REM-state activation will exhibit smaller cells, owing to the loss of extrinsic as well as intrinsic activation. DESIGN: We carried out a study comparing soma sizes in the MSs of kittens subjected to monocular deprivation (MD) + REM deprivation (RD) to two age-matched nonRD groups, MD ONLYs and MD MOMS (MD kittens living in their home cages). MEASUREMENTS AND RESULTS: Perikaryal outlines of 100 cells in each of the bilateral MSs were measured. As predicted, mean cell size in the MS connected to the patched eye of MD + RD kittens, but in neither of the control groups, was significantly smaller than in the MS afferented by the nonpatched eye. One-way ANOVAs comparing MS cell-size means from the same sides across groups were also significant, but the two MSs showed different results on post hoc tests. The ordering of MS cell-size means correlated significantly with a measure that aggregates the sources of activation reaching a particular MS and their durations. CONCLUSIONS: These results reveal that removal of REM-state activation during CNS development amplifies the plasticity processes generated when normal visual afferentation to central visual areas is interrupted. Our findings in the MS of the LGN indicate that during the usual operation of REM sleep, central visual-sensory sites receive intrinsic activation that, in the visual system, is additive and complementary to the stimulation obtained from extrinsic sources. In the course of early development, normative symmetrical activation of central visual areas during REM sleep may counterbalance plasticity changes caused either by absent or aberrant sensory stimulation.

Rapid eye movement sleep deprivation in kittens amplifies LGN cell-size disparity induced by monocular deprivation.

The abundance of rapid eye movement (REM) sleep in the neonatal mammal and its subsequent decline in the course of development, as well as the dramatic and widespread enhancement of CNS activity during REM sleep, led us to propose that this state plays a functional role in the normative physiological and structural maturation of the brain [54]. When, after 1 week of monocular deprivation (MD), a second week of MD was coupled with behavioral deprivation of REM sleep, the structural alteration in the visual system provoked by MD alone (interlaminar relay cell-size disparity in the lateral geniculate nucleus (LGN) was amplified. With the addition of REM deprivation during MD, the LGN cells connected to the surgically patched eye, which are smaller than normal after MD, became even smaller, whereas the LGN cells receiving input from the seeing eye, which display compensatory hypertrophy after MD, grew even larger. We believe that the interlaminar disparity effect widened because during REM deprivation, the already vision-compromised LGN cells associated with the patched eye also lose the ascending brainstem activation reaching them during the REM state. Loss of the two main sources of 'afference' by these LGN cells permits their seeing-eye LGN counterparts to gain even greater advantage in the competition for synaptic connections in cortex, which is reflected in the relative soma sizes of the LGN relay cells. It is likely that the relatively abundant REM state in early maturation provides symmetric stimulation to all LGN relay cells, irrespective of eye of innervation. The symmetric activation propagated from brainstem to LGN acts to 'buffer' abnormal, asymmetric visual input and, thereby diminishes the extreme, asymmetric structural alteration that results from MD in the absence of REM sleep. We conclude that REM sleep-generated CNS discharge in development has the effect of 'protecting' the CNS against excessive plasticity changes. This is consistent with the possibility that REM sleep plays a role in the genetically programmed processes that direct normative brain development.

A preliminary study of sleep in the ferret, Mustela putorius furo: a carnivore with an extremely high proportion of REM sleep.

Sleep and wakefulness were studied polygraphically in two adult male ferrets between the ages of 3 months and 2.5 years. Nine 24-hour recordings were obtained on different light/dark schedules, and one animal was also monitored for 3 days following administration of the serotonin synthesis inhibitor, parachlorophenylalanine. Stage scoring was accomplished utilizing criteria similar to those used in the cat. Certain modifications to the criteria were made to accommodate apparent differences in electrographic indicators of state. Mean daily percentages [+/- standard error of the mean (SEM)] for the major stages were: slow-wave sleep, 36.0 +/- 1.33; rapid eye movement (REM) sleep, 24.4 +/- 0.94; and wake, 39.4 +/- 1.17. Under laboratory conditions, ferrets spend over 60% of the time in sleep and 40.28% +/- 0.93% of total sleep time in REM sleep. The high amount of REM sleep is achieved by having a high number of REM sleep episodes rather than long REM periods. Sleep cycle length was computed in two ways: with long wake episodes removed, 16.7 +/- 0.4 minutes; and with all wake removed, 13.2 +/- 0.3 minutes. Sleep and waking indices in the ferret are compared to those in the cat and discussed with respect to predictions based on several constitutional variables. The high amount of REM sleep and the relative immaturity of the ferret at birth lends additional support for a functional role of REM sleep on ontogenetic development.

A functional role for REM sleep in brain maturation.

The biological function of REM sleep is defined in terms of the functions of neural processes that selectively operate during the REM sleep state. The high amounts of REM sleep expressed by the young during a period of central nervous system plasticity suggest that one function of REM sleep is in development. The phenomenon of activity-dependent development has been clearly shown to be one mechanism by which early sensory experience can affect the course of neural development. Activity-dependent development may be a ubiquitous process in brain maturation by which activity in one brain region can influence the developmental course of other regions. We hypothesize an ontogenetic function of REM sleep; namely, the widespread control of neuronal activity exerted by specific REM sleep processes help to direct brain maturation through activity-dependent developmental mechanisms. Preliminary tests of the hypothesis have been conducted in the developing feline visual system, which has long been known to incorporate information derived from visual experience in establishing neuronal connectivity. We find that suppression of REM sleep processes by an instrumental REM deprivation procedure results in a significant enhancement of the effects of altered visual experience by monocular occlusion. Bilateral brainstem lesions that selectively block the occurrence of ponto-geniculo-occipital (PGO) waves are sufficient to produce similar results. These data indicate that the propagation of phasic influences during REM sleep interacts with other processes subserving neural development. This source of influence appears not to derive from the environment but rather stems from an intrinsic source of genetic origin. Examination of the neural activity associated with PGO waves in the lateral geniculate nucleus reveals a distribution of facilitatory influence markedly different from that induced by visual experience. We conclude that REM sleep directs the course of brain maturation in early life through the control of neural activity.

Spontaneous activity in the thalamic reticular nucleus during the sleep/wake cycle of the freely-moving rat.

Neurons of the somatosensory thalamic reticular nucleus (TRN) were studied by extracellular recordings through the sleep/wake cycle in the unanesthetized, freely-moving rat. All electrophysiologically-identified TRN neurons expressed rhythmic patterns of discharge that altered with shifts in sleep/wake state. During slow wave (SW) sleep, neurons displayed spike-burst discharges in long trains followed by pauses. high-frequency oscillations in auto-correlograms in the spindle-frequency range (approximately 10 Hz) reflected the rhythm of interburst intervals within the trains whereas low-frequency oscillations (0.3-0.2 Hz) displayed the rhythm of intertrain intervals. During rapid eye movement (REM) sleep, a more continuous pattern of spike-burst discharges was prominent, resulting in absence of a detectable, low-frequency rhythm but persistence of spindle-frequency firing. At the transitions between SW and REM sleep, cell discharge was more tonic than during either sleep state and lacked a dominant rhythm. During the wake (AW) state, neurons fired in a single-spike mode that also lacked rhythmicity. Unlike their pattern of discharge, TRN neurons' mean rate of discharge did not distinguish sleep/wake state. The mean discharge rates were: SW, 18.4 +/- 1.3; REM, 17.4 +/- 1.2; AW, 22.3 +/- 2.1 (Hz +/- S.E.M.). Mean discharge rate during transitions from SW to REM sleep (28.6 +/- 2.1) was significantly higher, however, than during any sleep/wake state. Compelling evidence was lacking for segregation of TRN neurons into discrete populations according to absolute discharge rate. Neurons recorded simultaneously from the same electrode discharged synchronous trains of spike-bursts and pauses during SW sleep. This phenomenon may be related to generation of EEG slow waves.(ABSTRACT TRUNCATED AT 250 WORDS)

Cholinergic responsiveness of neurons in the ventroposterior thalamus of the anesthetized rat.

Acetylcholine has been implicated as an important neurotransmitter in the mechanisms of thalamic activation. Cholinergic mechanisms are thought to directly underlie the high level of excitability observed in thalamic relay neurons during waking and rapid eye movement sleep. We sought to determine if the cholinergic responsiveness of neurons in the ventroposterior nuclei of the thalamus in rat is consistent with this view. Neurons in the chloral hydrate-anesthetized rat were studied with extracellular recording and microiontophoretic application of cholinergic agents. In most cases (63% of 63 cells), the ejection of the agonist, carbachol, had no observable effect on spontaneous activity. Facilitation (25%), inhibition (8%) and inhibition followed by facilitation (3%) were also observed. Carbachol ejections that by themselves were ineffective in altering spontaneous activity proved capable, in 93% of 28 cells, of antagonizing the uniformly facilitatory responses produced by glutamate ejection. The putative M1-selective, cholinergic agonist, McN-A-343, was also ineffective alone in altering spontaneous activity in the majority of cases (74% of 27 cells) and produced only inhibitory responses in the remaining seven neurons studied. Interacting applications of McN-A-343 and glutamate resulted, in all cases, in antagonism of glutamate facilitation (N = 12). The various responses to applied cholinergic agonists were all capable of being antagonized by muscarinic receptor-blocking agents. Both the high proportion of inhibitory responses and the antagonism of glutamate facilitatory responses suggest that ventroposterior neurons in the rat differ from other thalamocortical relay neurons in the rat and cat with regard to cholinergic responsiveness. Additionally, the lack of predominantly facilitatory responding renders it unlikely that cholinergic mechanisms directly underlie increases in excitability of ventroposterior neurons observed during waking and rapid eye movement sleep.

Cholinergic modulation of responses to glutamate in the thalamic reticular nucleus of the anesthetized rat.

Neurons in the thalamic reticular nucleus (TRN) of the chloral hydrate-anesthetized rat were studied with extracellular recording and microiontophoretic application of cholinergic agents. In most cases (63%), the ejection of the agonist, carbachol, had no observable effect on spontaneous activity, and in an additional 33% of cases was observed to inhibit discharge rate. Carbachol ejections with identical current and duration parameters proved capable of antagonizing the uniformly facilitatory responses produced by glutamate ejection in these same cells. The muscarinic nature of cholinergic effects was documented by scopolamine's specific antagonism of the responses. The muscarinic antagonists, pirenzepine and AF-DX-116, both diminished the effects of carbachol. Application of muscarinic agonists, such as McN-A-343 and oxotremorine-M, yielded qualitatively the same results as carbachol, though, with current as a criterion, oxotremorine-M was slightly more and McN-A-343 much less potent than carbachol. The functional implications of cholinergic modulation of the facilitatory inputs to TRN are discussed, with particular emphasis on the role of acetylcholine and the TRN in the sleep/wake-related activity of thalamic neurons.

The cholinergic influence upon rat dorsal lateral geniculate nucleus is dependent on state of arousal.

Several lines of evidence suggest a role for acetylcholine (ACh) in mediating the effects of state of arousal on transfer of visual information through the lateral geniculate nucleus (LGN). Local application of cholinergic agonists to geniculate relay cells in anesthetized cats and rats produces predominantly facilitatory effects. This indicates that presynaptic release of ACh may be responsible for the increased excitability of LGN relay cells that is observed during waking and REM sleep. In this study in rats we have examined the influence of cholinergic agonists applied during the 3 natural states of arousal: waking, slow-wave (SW) sleep and rapid eye movement (REM) sleep. Pharmacological agents were iontophoretically administered to identified, single cells in head-restrained, unanesthetized rats free to sleep and wake. Application of cholinergic agonist produced state-dependent differences in response in all geniculate relay-cells studied. During both waking and REM sleep, a facilitatory response was always observed, whereas in SW sleep responses were of three types: no effect (62%), inhibition (24%), and biphasic inhibition followed by facilitation (14%). All response types were antagonized by scopolamine. In contrast to the qualitatively different state-dependent effects of cholinergic agonists, response to application of glutamate, with quantitative variations, was uniformly facilitatory in all states, though responses in SW sleep tended to be lower in magnitude. The effects of gamma-aminobutyric acid (GABA), glycine, and serotonin were inhibitory in all states. These data are consistent with the suggested role of ACh in mediation of increased relay-cell excitability during REM sleep and waking. Our findings, however, also indicate that in the transition from SW sleep to REM or waking, local release of ACh is not solely responsible for alterations in cell excitability.

Sleep state-specific neuronal activity in rat dorsal lateral geniculate nucleus is not altered by local serotonin and norepinephrine depletion.

The relay cells in dorsal lateral geniculate nucleus (dLGN) represent one among many populations of neurons throughout the neuraxis that display systematic alteration of spontaneous rate and pattern of discharge concurrent with change in state of arousal. Both noradrenergic (NE) and serotonergic (5-HT) systems innervate dLGN and are implicated in sleep-wake control mechanisms. Our study was designed to test the influence of these systems upon sleep state-related multiple unit activity in the dLGN. Two monoamine neurotoxins, 5,7-dihydroxytryptamine and 6-hydroxydopamine, were injected locally into dLGN to destroy NE and 5HT afferents individually and in combination. In three separate treatment groups, mean monoamine concentrations in dLGN were reduced in relation to the contralateral, vehicle-injected, control dLGN to: 1) NE-17%, 5HT-14%; 2) NE-46%, 5HT-28%, and 3) NE-6%, 5HT-77%. In no case was chronic sleep state-related cell activity in dLGN significantly altered. We conclude that afferent monoaminergic inputs are not critically related to the mechanisms underlying normative shifts in sleep state-related neuronal activity in dLGN.

PGO waves and insomnia in PCPA-treated rats.

The serotonin-depleting drug, parachlorophenylalanine (PCPA), in a dosage of 300 mg/kg, was administered to rats in an effort to test the hypothesis that altered distribution of PGO waves following drug treatment may be responsible for the sleep disruption and consequent sleep loss that accompany decreased serotonin levels. Consistent with the hypothesis, we found that the greater the proportion of PGO waves that precede spontaneous arousals, the greater the reduction in slow wave sleep. However, inconsistent with the hypothesis, we found that the decrease in sleep did not result from an increase in the number of arousals. Further, though an increase in the proportion of waking waves always accompanied a rise in wake time, the two variables were negatively correlated. These data do not support a PGO wave/arousal hypothesis to account for the decrease in sleep following PCPA treatment in the rat. Rather, the findings tend to implicate an alteration in the mechanisms of arousal linked to serotonin depletion.

Evidence for a role of delta sleep-inducing peptide in slow-wave sleep and sleep-related growth hormone release in the rat.

To examine the role of delta sleep-inducing peptide (DSIP) in sleep-related growth hormone (GH) release, male rats were deprived of sleep for 4 hr by placing them on a slowly rotating wheel. Sleep deprivation by this method caused a significant increase in GH release, as indicated by the increase in plasma GH concentrations (P less than 0.01), and also in the amount of slow-wave sleep (SWS) (P less than 0.001) above initial values after removal of the animals from the rotating wheel. These increases were blocked by microinjection into the third cerebral ventricle of highly specific antiserum to DSIP. In control rats receiving an equal volume of normal rabbit serum, the significant increase in plasma GH as well as SWS remained after removal of the rats from the wheel. The increased release of endogenous DSIP in the sleep-deprived animals may have caused an increase in SWS as well as plasma GH. Since DSIP increases plasma GH after its injection into the third cerebral ventricle and since passive immunization against DSIP blocks the increase in SWS and GH release that follows the 4 hr of sleep deprivation, the results suggest that DSIP can be a physiological stimulus for sleep-related GH release as well as for the induction of SWS.

A method for concurrent local intracranial drug infusion and electrophysiological recording.

A method employing a cannula-guidetube system with attached electrodes is described that permits infusions into discrete brain areas and simultaneous recording of multiunits or single units. This method can be used to determine the immediate local effects of drug infusion, and also to acquire electrophysiological feedback information in the process of placement and identification of cell populations at the injection site.