GABAergic deafferentation hypothesis of brain aging and Alzheimer's disease revisited.

Considering the mechanisms responsible for age- and Alzheimer's disease (AD)-related neuronal degeneration, little attention was paid to the opposing relationships between the energy-rich phosphates, mainly the availability of the adenosine triphosphate (ATP), and the activity of the glutamic acid decarboxylase (GAD), the rate-limiting enzyme synthesizing the gamma-amino butyric acid (GABA). Here, it is postulated that in all neuronal phenotypes the declining ATP-mediated negative control of GABA synthesis gradually declines and results in age- and AD-related increases of GABA synthesis. The Ca2+-independent carrier-mediated GABA release interferes with Ca2+-dependent exocytotic release of all transmitter-modulators, because the interstitial (ambient) GABA acts on axonal preterminal and terminal varicosities endowed with depolarizing GABA(A)-benzodiazepine receptors; this makes GABA the "executor" of virtually all age- and AD-related neurodegenerative processes. Such a role of GABA is diametrically opposite to that in the perinatal phase, when the carrier-mediated GABA release, acting on GABA(A)/chloride ionophore receptors, positively controls chemotactic migration of neuronal precursor cells, has trophic actions and initiates synaptogenesis, thereby enabling retrograde axonal transport of target produced factors that trigger differentiation of neuronal phenotypes. However, with advancing age, and prematurely in AD, the declining mitochondrial ATP synthesis unleashes GABA synthesis, and its carrier-mediated release blocks Ca2+-dependent exocytotic release of all transmitter-modulators, leading to dystrophy of chronically depolarized axon terminals and block of retrograde transport of target-produced trophins, causing "starvation" and death of neuronal somata. The above scenario is consistent with the following observations: 1) a 10-month daily administration to aging rats of the GABA-chloride ionophore antagonist, pentylenetetrazol, or of the BDZ antagonist, flumazenil (FL), each forestalls the age-related decline in cognitive functions and losses of hippocampal neurons; 2) the brains of aging rats, relative to young animals, and the postmortem brains of AD patients, relative to age-matched controls, show up to two-fold increases in GABA synthesis; 3) the aging humans and those showing symptoms of AD, as well as the aging nonhuman primates and rodents--all show in the forebrain dystrophic axonal varicosities, losses of transmitter vesicles, and swollen mitochondria. These markers, currently regarded as the earliest signs of aging and AD, can be reproduced in vitro cell cultures by 1 microM GABA; the development of these markers can be prevented by substituting Cl- with SO4(2-); 4) the extrasynaptic GABA suppresses the membrane Na+, K+-ATPase and ion pumping, while the resulting depolarization of soma-dendrites relieves the "protective" voltage-dependent Mg2+ control of the N-methyl-D-aspartate (NMDA) channels, thereby enabling Ca2+-dependent persistent toxic actions of the excitatory amino acids (EAA); and 5) in whole-cell patch-clamp recording from neurons of aging rats, relative to young rats, the application of 3 microM GABA, causes twofold increases in the whole-cell membrane Cl- conductances and a loss of the physiologically important neuronal ability to desensitize to repeated GABA applications. These age-related alterations in neuronal membrane functions are amplified by 150% in the presence of agonists of BDZ recognition sites located on GABA receptor. The GABA deafferentation hypothesis also accounts for the age- and AD-related degeneration in the forebrain ascending cholinergic, glutamatergic, and the ascending mesencephalic monoaminergic system, despite that the latter, to foster the distribution-utilization of locally produced trophins, evolved syncytium-like connectivities among neuronal somata, axon collaterals, and dendrites, to bidirectionally transport trophins. (ABSTRACT TRUNCATED)

GABAergic deafferentation hypothesis of brain aging and Alzheimer's disease; pharmacologic profile of the benzodiazepine antagonist, flumazenil.

Recent experiments have shown that: 1) A chronic 10 month daily administration to rats of the benzodiazepine (BDZ) receptor antagonist, flumazenil (FL; 4 mg/kg in drinking water), from the age of 13 through 22 months, significantly retarded the age-related loss of cognitive functions, as ascertained by the radial arm maze tests conducted two months after FL withdrawal. 2) An equal number of 8 rats died in the control and FL-treated group before the behavioral tests were completed and the animals were sacrificed; the life span of the FL-treated 8 rats equaled 24.0 (+/- 0.6 SEM) months, while that of the control 8 rats equaled 22.3 months (+/- 0.7 SEM), and the group difference was marginally significant (p = 0.04 Mann-Whitney test). 3) In rats sacrificed 3 months after FL withdrawal and behavioral testing, the protective action of FL, relative to age-matched controls, was revealed by a significant reduction in the age-related loss of neurons in the hippocampal formation. 4) In the time period of 3 months between the drug withdrawal and sacrificing of the animals, stress experienced by the aging rats during behavioral testing, related to excessive daily handling of the animals and partial food deprivation to motivate them to perform in the radial arm maze, apparently had excitotoxic effects on the hippocampal neurons, as indexed by the presence of 30% neurons in a state of moderate pyknosis found both in the FL group and the age-matched controls. In the 6 months "young" control group, the number of pyknotic neurons equaled only 3.5%. It was concluded that the drug withdrawal and stress of behavioral testing unleashed the previously FL-controlled age-related degeneration. On the basis of these results and the literature, showing that the tone of the GABAergic system increases with age, and particularly in Alzheimer's disease (AD), the hypothesis of brain aging was formulated. It postulates that in mammals, with growing age, and prematurely in humans with AD, the increasing tone of the BDZ/GABAergic system interferes with antero- and retrograde axonal transport through a chronic depolarizing block of preterminal axon varicosities of the ascending aminergic and cholinergic/peptidergic systems, which are indispensable for normal metabolic/trophic glial-neuronal relationships. Such a state leads to discrete anatomic deafferentation of forebrain systems, and particularly of the neocortex, where block of the anterograde axonal transport results in induction of the cortical mRNA responsible for synthesis of the beta-amyloid precursor protein (beta APP). The simultaneous block of retrograde transport from chronically depolarized preterminal axon varicosities may account for toxic accumulation in cortex of the nerve growth factor (NGF) and other trophins, without which the basal forebrain cholinergic neurons degenerate. The general pharmacologic profile of FL has been discussed on the basis of FL administration to animals and healthy and diseased humans. This profile shows that FL: 1) increases brain metabolic functions; 2) reduces emotional responses, thereby stabilizing the functions of the autonomic system in both humans and animals challenged by adverse environmental stimuli; 3) improves cognitive and coordinated motor functions in both humans and animals; 4) uniquely combines anxiolytic, vigilance and cognitive enhancing, i.e. nootropic, properties, which may, in part, stem from FL-induced emotional imperturbability (ataraxy); 5) facilitates habituation of healthy humans and animals to novel but inconsequential environmental stimuli, and promotes non-aggressive interactions among animals; 6) in single i.v. doses, and administered chronically to humans, FL has antiepileptic actions in the Lennox-Gastaut syndrome and other forms of epilepsy characterized by "spike-and-dome" EEG patterns; these actions are likely to depend on FL's disinhibition of the serotonin system; 7) administered in single i.v...

Chronic administration of flumazenil increases life span and protects rats from age-related loss of cognitive functions: a benzodiazepine/GABAergic hypothesis of brain aging.

Under barrier condition and with ad lib access to food and water, 20 Fischer-344 rats were chronically treated for 10 months with the benzodiazepine (BDZ) antagonist, flumazenil (FL; 4 mg/kg/day in drinking water acidified to pH = 3.0), beginning at the age of 13 months, while the group of 20 control age-matched rats received plain acidified water. The life span of the first 8 deceased rats treated with FL was significantly longer than that of the first 8 deceased rats in the age-matched control group. In tests for spontaneous ambulation and exploratory behavior in the Holeboard apparatus, conducted during the 3rd and the 8th month of treatment, the FL group, relative to controls, had significantly higher scores for the ambulation and exploratory behavior. In tests for unrewarded spontaneous alternation in the T maze, conducted at days 7, 39, 42, and 47 through 54 after drug withdrawal, i.e., at the age of 24-25 months, the FL-exposed group, compared to age-matched controls, showed a significantly higher percent of alternating choices, a behavior that was statistically comparable to that of the "young" 6-month-old controls. In the Radial Maze tests conducted 2 months after drug withdrawal, the FL group made significantly less "working memory" errors and "reference memory" errors, relative to the age-matched 25-month-old control group, a performance that was comparable to that of the young 7-month-old control group. In conclusion, chronic FL significantly protected rats from age-related loss of cognitive functions. It is postulated that the age-related alterations in brain function may be attributable to the negative metabolic/trophic influences of the "endogenous" benzodiazepine (BDZ) ligands and/or those ingested with food. A BDZ/GABAergic hypothesis of brain aging has been formulated which assumes that age-related and abnormally strong BDZ/GABAergic influences promote neurodegeneration by suppressing trophic functions of the aminergic and peptidergic neurons through opening of chloride channels in soma membrane and axon terminals, causing excessive hyperpolarizing and depolarizing inhibition, respectively. The review of human clinical and animal data indicates that FL has nootropic actions by enhancing vigilance cognitive and habituation processes.

A low-noise preamplifier for multisite recording of brain multi-unit activity in freely moving animals.

A novel FET instrumentation amplifier is described which, as compared to most traditional operational FET preamplifiers, is characterized by an about 7-10 times lower intrinsic electronic noise and a higher common mode rejection. This allows discrimination of single units from multi-unit recording, even if the action potential amplitudes are as small as 20-30 microV and the units are located more than 100 microns away from the electrode tips. Such a distant and chronic recording may be expected to reduce the possibility of mechanical interference with functions of neuronal membrane and its immediate environment, and may be suitable for studying changes in functional connectivities among neurons during the animal's behavior and learning.

Benzodiazepines inhibit neutrophil chemotaxis and superoxide production in a stimulus dependent manner; PK-11195 antagonizes these effects.

Diazepam, which binds both central (neuronal) and peripheral (non-neuronal) benzodiazepine binding sites, and Ro5-4864, a ligand selective for benzodiazepine peripheral binding sites (PBS), both inhibited the FMLP induced chemotaxis in human neutrophils at concentrations as low as 10(-8) M. A selective peripheral benzodiazepine antagonist, PK-11195 (10(-5) M), partially reversed the benzodiazepine inhibition of chemotaxis. Diazepam also inhibited the superoxide production induced by FMLP, NaF, and A23187, but not that induced by PMA whose stimulant action was insensitive even to 10(-4) M diazepam. The FMLP-induced superoxide production was most sensitive to diazepam inhibition (ID50 = 2.25 x 10(-6) M diazepam); the effect of NaF was slightly less sensitive (ID50 = 1.34 x 10(-5) M diazepam); and the effect of A23187 was least sensitive as it was suppressed only at 10(-4) M diazepam concentrations. Like diazepam, Ro5-4864 inhibited the FMLP-induced superoxide production, and PK-11195 (10(-5) M) significantly antagonized both diazepam and Ro5-4864 inhibition. Binding studies showed the presence of a saturable benzodiazepine 'peripheral' type binding site (PBS) on human neutrophils with a Kd of 1.2 +/- 0.06 x 10(-8) M (+/- SEM), and a Bmax of 1028 +/- 86.2 fmol/10(6) cells (+/- SEM) for [3H]Ro5-4864; the binding was displaceable by PK-11195, Ro5-4864 and diazepam but not by clonazepam.

Chronic administration of flumazenil (Ro 15-1788) enhances non-appetitive exploratory behavior of rats.

The effects of chronic administration of the benzodiazepine receptor antagonist, flumazenil (Ro 15-1788; 4 mg/kg/day for 14 days in drinking water) on the performance of adult rats in the 12-arm radial maze were studied. Relative to controls, the animals treated with flumazenil showed an increase (P less than 0.002) in non-appetitively motived exploratory behavior, so called because it occurred in 88% of instances in non-baited alleys, facing the well-illuminated "enriched environment" of the center of the room, as opposed to the baited alleys, facing the "dull" corner of the room. This behavior emerged between day 5 and 7 of treatment with the drug, it continued to increase over the period of treatment with drug (P less than 0.002), and reached its peak at day 3, after withdrawal of the drug (P less than 0.008; a longer duration was not investigated). The occurrence of non-appetitively motivated exploratory behavior was inversely correlated with the scores for urination/defecation (P less than 0.003) and, therefore, most likely reflected the anxiolytic action of flumazenil. During treatment with drug or vehicle, the control and the drug groups made comparable numbers of "working memory" errors (P = 0.17). However, upon withdrawal of drug and introduction of alley gates (to confine the animal for 10 sec to the center platform, after an alley was explored), the working memory errors of the rats exposed to the drug, remained unchanged (P = 0.35), relative to the preceding three trials, while the performance of the control group was disrupted, as shown by an increase in the numbers of errors (P less than 0.004). At day seven of treatment with drug, the emergence of exploratory behavior was associated with an increased density and/or affinity of benzodiazepine receptors in cortex, hippocampus and brain stem, while three days after withdrawal of drug, when the exploratory behavior reached its peak, there was a reduction in GABA-enhanced binding of [3H]flunitrazepam in the cortex.

Chronic Ro 15-1788 treatment increases REM sleep in rats.

Administration of Ro 15-1788, a benzodiazepine antagonist (3.6 mg/kg/day in drinking water for 14 days), increased total sleep and rapid eye movement (REM) sleep in rats. Standard six-hour EEG recording periods were obtained on day 0, 1, 3, 7, 10, 14, as well as 24 and 72 hours following withdrawal. Enhanced REM sleep reached significance on day 7 of continuous drug treatment and remained significantly increased on day 10 and 14, as well as at 24 and 72 hours following drug withdrawal. The present data show that chronic administration of Ro 15-1788 increases total sleep time due to increases in REM sleep. The actions of Ro 15-1788 presumably occur through either adenosinergic or cholinergic mechanisms.

Chronic exposure to flumazenil: anxiolytic effect and increased exploratory behavior.

The aim of the present study was to define the behavioral correlates of chronic exposure of adult rats to flumazenil (4 mg/kg/day X 21 days in drinking water). In the holeboard test, performed on day 13 of drug treatment, the animals showed a significantly greater interest for the holes under which objects were placed than for the holes without objects (p less than 0.03), while there was no such difference in the control group. In the plus-maze test, the flumazenil-treated animals spent significantly more time on open arms and left less fecal boluses than the controls when tested in the third week of treatment and 24 hours after flumazenil withdrawal. In the drinking-punishment test, conducted on days 3, 6 and 10 after drug withdrawal, the drug-exposed animals, following shock experience, did not significantly alter their unpunished drinking in subsequent trials, while the control rats significantly reduced (p less than 0.003) their unpunished drinking. Also, the punished drinking revealed a significant "anticonflict" effect of prior exposure to flumazenil (p less than 0.006) which was still observed 6 days after drug withdrawal. There were no group differences in the home-cage food and water consumption during flumazenil treatment; also, the drug treatment had no effect on nociceptive threshold. In summary, chronic treatment with a benzodiazepine receptor antagonist, flumazenil, increased exploratory activity and had a lasting anxiolytic effect.

Chronic flumazenil (Ro 15-1788) facilitates acquisition and retention of a swim-escape response in rats.

Since chronic flumazenil treatment was previously found to stimulate exploratory behavior in rodents, the aim of this study was to test the effect of chronic exposure to flumazenil on acquisition and retention of escape behavior. Adult rats were treated with flumazenil (Ro 15-1788; 4 mg/kg/day in drinking water) for 21 days (experiment 1) and for 17 days (experiment 2). In experiment 1 (a round water tank with one escape rope) conducted 24 h after drug/vehicle withdrawal, the time the animals needed to resolve a swim-escape task was significantly shorter in the drug group, compared to the controls. In the retention trial, 24 h later, the control group matched the performance of the drug group. In experiment 2, a water T-maze was used which was equipped with two ropes, one anchored to the bottom and the other unanchored and therefore was more difficult to climb. On day 14 of flumazenil/vehicle treatment, there were no differences between the groups in the time needed to escape from the maze. However, on day 15 and 16 of drug/vehicle treatment, the drug group made highly significant progress, while the control group showed no improvement of the escape behavior. The possible mechanisms of flumazenil-induced facilitation of escape behavior have been discussed.

Chronic exposure to Ro 15-1788: differential effect on flunitrazepam binding to cortex and hippocampus.

The time course of changes in specific [3H]flunitrazepam binding following 2 weeks of treatment with the benzodiazepine antagonist Ro 15-1788 (2.7 and 4 mg/kg per day in drinking water) was studied in the rat neocortical and hippocampal synaptosomal membranes. Such a treatment produced regional increases in the density of benzodiazepine sites, which remained for up to 24 and 48 h after drug withdrawal in the hippocampus and cortex, respectively; the dissociation constant (Kd) was unchanged. In addition, a significant reduction in GABA enhanced [3H]flunitrazepam binding to cortical, but not to hippocampal, membranes from Ro 15-1788-treated animals was found 72 h after drug withdrawal. These data show that there is a regional difference in responses of the GABA/benzodiazepine receptor complex following chronic in vivo exposure to Ro 15-1788 and that, beside the increases in the maximal binding (Bmax) of [3H]flunitrazepam the coupling between the GABA and the benzodiazepine recognition sites was also affected.

Adenosine presynaptic inhibition and transmitter "spillover": a new hypothesis of etiopathogenesis of narcolepsy.

Neurobiologic and clinical evidence has been discussed in order to propose a new hypothesis explaining the precipitous nature of narcoleptic attacks. It is postulated that narcoleptic episodes are triggered by a surge in the tone of the arousal system which temporarily overcomes the abnormal tonic inhibitory influences of adenosine on presynaptic terminals of the arousal system. As a result, abnormally high levels of accumulated transmitters "spillover" onto supersensitive postsynaptic receptors both in the brain and spinal cord. Such a state reduces the tone of the skeletal muscles and blocks the thalamo-cortical association system, causing a hypnagogic state incompatible with adaptive and cognitive functions. An agent selectively blocking A1 receptors would constitute the most appropriate treatment of narcolepsy. In theory, the hereditary predisposition toward narcolepsy could be corrected by perinatal treatment with an agonist of A1 receptors, thus causing an enduring down-regulation of the genome expression that regulates the ontogeny and proliferation of the A1 receptors.

Animal models of chronic anxiety and "fearlessness".

Three behavioral animal models have been described: a feline and a rodent model of chronic anxiety, and a rodent model of "fearless" behavior. The models have been obtained by pre- or perinatal exposure to diazepam (DZ) or RO 15-1788 which produced enduring postnatal deficits or enrichment, respectively, of brain benzodiazepine (BDZ) receptors. The receptor-deficient one-year-old cat progenies showed hyperarousal, unabated restless behavior, delayed acquisition of instrumentally conditioned behavior, bizarre escape responses and absence or reduced alpha-like EEG activity. The receptor-deficient rat progencies, studied at the age of 5-6 months, showed a reduction of time spent in deep slow wave sleep, and inability to habituate to novel environment, such as the radial arm maze. In the maze, the behavior of these progenies was characterized by delayed and incomplete exploratory activity often terminated by sudden escape, numerous fecal deposits and significantly more frequent than normal errors of "working memory." On the other hand, in all aspects, the receptor-enriched progenies were superior to the control animals as well as to the receptor-deficient group, particularly when the animals were challenged by novel and "intimidating" visual and/or auditory stimuli. In addition, 12 out of 51 receptor-deficient rats reared for 18 months developed mammary fibroadenomas, while no such tumors were found in the group of 44 vehicle-exposed control animals. Increased density and affinity of BDZ brain receptors was also observed after adult rats were treated with RO 15-1788 administered water for 7 or 14 days.

Perinatal upregulation of benzodiazepine receptor ontogenesis: "fearless" and more efficient goal-directed behavior of adult rat progenies.

Pregnant and subsequently lactating rats had ad lib access to drinking water which contained either a benzodiazepine antagonist, Ro 15-1788 or diazepam (DZ). On the average, the rats consumed 2.9 mg/kg/day of Ro 15-1788 or 5.3 mg/kg/day of diazepam over the time period of 3 weeks, from gestation day 14 through postpartum day 14. The control group consumed equivalent volumes of the drug vehicle in water. While Ro 15-1788 had no apparent toxic effects, the numbers of viable pups in the DZ group were significantly reduced. The mean weight of the viable pups, and their gross behavior were not different in all three groups. However, the fully mature 4.5-month-old male progenies exposed to Ro 15-1788 were much more efficient in the radial arm maze test than the control or the diazepam-exposed animals; they rapidly habituated to the novel environment, their exploratory activity was uninhibited by distracting visual and auditory stimuli, they made fewer "working memory" errors in collecting baits, had a much better control over their emotional responses and the autonomic nervous system, as shown by very low defecation/urination scores, and, at the age of 5 months, they had a significant (66%) increase in the density of benzodiazepine receptors in the hippocampal formation, as compared to the control or the diazepam-exposed progenies. In conclusion, the upregulation of benzodiazepine receptor ontogenesis is retained in adult animals and resulted in improved "working memory" and better control over emotional responses that were particularly evident when the animals were challenged by novel and "intimidating" environmental stimuli.

Prenatal diazepam: chronic anxiety and deficits in brain receptors in mature rat progeny.

Pregnant rats were treated with daily doses (5.0 to 7.5 mg/kg, SC) of the benzodiazepine (BDZ) receptor agonist, diazepam (DZ) between day 15 through day 20 of gestation, i.e., during ontogenesis of BDZ receptors in an attempt to alter their development. In the radial arm maze, 6 months old rat progeny showed behavioral anomalies and deficient exploratory behavior. One plausible interpretation of their poor performance is a deficit in short-term spatial "working memory." However, a more detailed evaluation of their behavior suggests a chronic state of hyperarousal/anxiety and thus, poor focus of attention on the task at hand. In addition, the number of brain BDZ receptors in the thalamus, at the age of one year, was significantly reduced. Hence, prenatal exposure to DZ has enduring and detrimental effects on brain function.

Prenatal exposure to diazepam: late postnatal teratogenic effect.

During the last 5 days of gestation, pregnant Sprague-Dawley rats were treated daily with SC diazepam (DZ) injections (average dose 6 mg/kg of body weight). The control pregnant rats were treated with corresponding volumes of the vehicle. The progenies were examined for the occurrence of neoplastic and non-neoplastic lesions for up to 20 months. Although there were no early postnatal effects of DZ, 13 neoplasms developed in the 52 DZ-exposed rats (12 mammary fibroadenomas and one uterine sarcoma), while there were not such lesions in 44 control rats (p less than 0.001, Fisher's exact test). The non-neoplastic lesions in the DZ-exposed group, such as infections, arteriosclerosis etc., were also significantly greater in magnitude and incidence, compared to control animals (p = 0.007, Wilcoxon Rank Sum test). Also, the DZ-exposed rats had a significantly lower titre of the plasma immunoglobulin G (IgG), higher levels of white blood cells and lower hematocrit values than the control animals. DZ is known to bind with high affinity to both central nervous system and non-neuronal receptors present in peripheral organs and blood cells, such as monocytes. The monocyte receptors appear to be critical for chemotaxis induced by many agents affecting the normal function of the immune system including antineoplastic defense. We have previously shown that prenatal DZ suppresses ontogenesis of brain benzodiazepine receptors as tested in adult 12 months old cat and rat progenies. If the ontogenesis of "peripheral" receptors is also suppressed, this would provide a plausible explanation for teratogenic effect of DZ.

Enduring effects of prenatal diazepam on the behavior, EEG, and brain receptors of the adult cat progeny.

Pregnant cats were treated with a benzodiazepine receptor agonist, diazepam (DZ; Valium; average dose of 0.4 mg/kg/day, i.m.) between day 20 through day 53 of gestation in an attempt to alter the ontogenesis of the benzodiazepine receptors. As tested in fully developed one-year-old progenies, prenatal exposure to DZ resulted in a behavioral syndrome of hyperactivity, aggressiveness, behavioral signs of chronic anxiety, inability to habituate to novel environment, suppression or absence of the reward-induced alpha-like (7 to 11 Hz) electroencephalographic patterns during operantly conditioned behavior (bar pressing for 1 ml of milk reward), and deficits in the numbers of benzodiazepine receptors in the hypothalamus (-47%), fronto-orbital cortex (-33%) and postcentral cortex (-19%).

Prenatal exposure to diazepam results in enduring reductions in brain receptors and deep slow wave sleep.

After prenatal exposure to diazepam (Valium), mature rats at 4 months of age displayed slow wave sleep (SWS) electroencephalographic patterns indicating impaired synchronization and SWS mechanisms. These animals spent a much greater portion of their SWS in the lighter SWS I, as compared to the control group which showed a predominance of the deeper SWS II. At one year of age, the diazepam-exposed rats had much fewer diazepam-specific binding sites in the thalamus than the vehicle-exposed controls. These results provide first evidence for a physiological role for benzodiazepine receptors by showing that prenatal exposure to diazepam has an enduring and detrimental effect on their ontogenesis and sleep mechanisms.

Sleep and purposive behavior: inverse deviations from randomness of neuronal firing patterns in the feline thalamus. A new form of homeostasis?

In behaving cats trained to press a bar for small aliquots of milk reward, single neuronal firing patterns were monitored from the nucleus reticularis (NR) thalami during bar bressing (BP), subsequent quiet wakefulness with EEG spindles (S- QW ), grooming behavior (GR) and slow-wave sleep (SWS). The temporal patterns in the neuronal spike trains were analyzed using a non-parametric method based on relative relations between sequential spike intervals. The deviations of pattern occurrences from the random model were quantified. During BP, specific patterns occurred much more often while others occurred much less often than predicted by the random model. Patterns that were dominant during BP, were selectively suppressed or virtually eliminated during S- QW , GR and SWS, despite the increased firing rate; and, vice versa, patterns that were suppressed below chance level during BP, became dominant during S- QW , GR and SWS. The magnitudes of these inversions of the statistical distribution of patterns were not random but graded and positively correlated, thus indicating that they were homeostatically controlled. Since the inversions were already evident shortly after the satiated ceased bar pressing, they may be related to the 'need' for sleep. On the basis of the known mechanisms of pattern generation and changes in receptors for putative transmitters, it was postulated that the inversions of pattern distribution are related to the recuperative function of SWS, i.e. resensitization of receptors that had been desensitized during the animal's stereotypic BP performance. The NR and other neuronal ensembles seem to constitute an oscillatory system with two modes of reciprocal connectivities : one is supporting wakefulness and emission of specific firing patterns, and the other is incompatible with wakefulness and instead is associated with inversion of statistical distribution of firing patterns and recuperative function of SWS.

Sleep-wakefulness: inverse deviation from randomness of neuronal firing patterns in the feline thalamus. A new form of homeostasis?

During stereotypic goal-directed behavior, neurons in the feline nucleus reticularis thalami emitted specific temporal patterns, while other patterns occurred much less often than predicted by the random model. During subsequent slow wave sleep, the mean firing rate increased, but the patterns that were emitted during behavior were eliminated or suppressed far below chance level, while those that were previously suppressed became dominant.