Impact of anesthetic regimen on the respiratory pattern, EEG microstructure and sleep in the rat model of cholinergic Parkinson's disease neuropathology.

OBJECTIVES: We hypothesized that the impact of distinct anesthetic regimens could be differently expressed during anesthesia and on post-anesthesia sleep in the neurodegenerative diseases. Therefore, we followed the impact of ketamine/diazepam and pentobarbital anesthesia in a rat model of the severe Parkinson's disease cholinergic neuropathology on the electroencephalographic (EEG) microstructure and respiratory pattern during anesthesia, and on the post-anesthesia sleep. METHODS: We performed the experiments on adult, male, spontaneously breathing Wistar rats chronically instrumented for sleep recording. The bilateral pedunculopontine tegmental nucleus (PPT) lesion was done by ibotenic acid microinfusion. Following postoperative recovery, we recorded sleep for 6h, induced anesthesia 24h later using ketamine/diazepam or pentobarbital, and repeated sleep recordings sessions 48h and 6days later. During 20min of each anesthesia we recorded both the EEG and respiratory movements. For sleep and EEG analysis, Fourier analysis was applied on 6-h recordings, and each 10-s epoch was differentiated as a state of wakefulness (Wake), non-rapid eye movement (NREM) or rapid eye movement (REM). Additionally, the group probability density distributions of all EEG frequency band relative amplitudes were calculated for each state, with particular attention during anesthesia. For respiratory pattern analysis we used Monotone Signal Segments Analysis. The PPT lesion was identified through nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase histochemistry. RESULTS AND CONCLUSIONS: Our data show that the ketamine/diazepam anesthetic regimen in the PPT-lesioned rats induces more alterations in the EEG microstructure and respiratory pattern than does the pentobarbital anesthesia. In addition, the equal time required to establish an anesthetized state, and the long-term effect on post-anesthesia sleep in the PPT-lesioned vs. control rats suggest this anesthetic regimen as potentially more beneficial both for anesthesia induction and for post-anesthesia sleep in the surgical procedures of the elderly, and Parkinson's, and Alzheimer's patients.

High volume microinfusion suppresses local astrocyte response within nucleus basalis of rat.

Our study investigates the impact of different volume sham control and excitotoxin microinfusions in vivo on local reactive astroglial response within rat nucleus basalis (NB). We followed the effects of unilateral 200, 100, and 50 nL of sham-control (phosphate buffer PBS) versus ibotenic acid (IBO) microinfusions, mechanical NB lesion (10 µL Hamylton syringe needle positioned into NB for 5 min), or physiological control (intact brain), on the local reactive astroglial response within the NB site, by immunoreactivity against glial fibrillary acidic protein (GFAP). NB lesions were identified by NADPHdiaphorase histochemistry. Local astrocytes responses within NB were suppressed by both high volume microinfusions, PBS and IBO (200 and 100 nL) versus mechanical lesion. Our study has proved, for the first time, the volume of microinfusion as critical for any selective pharmacological stimulation or lesion in vivo, and suggest the microinfusion volume less than 50nL as protective for physiological astroglial reactivity.

Independent complexity patterns in single neuron activity induced by static magnetic field.

We applied a combination of fractal analysis and Independent Component Analysis (ICA) method to detect the sources of fractal complexity in snail Br neuron activity induced by static magnetic field of 2.7 mT. The fractal complexity of Br neuron activity was analyzed before (Control), during (MF), and after (AMF) exposure to the static magnetic field in six experimental animals. We estimated the fractal dimension (FD) of electrophysiological signals using Higuchi's algorithm, and empirical FD distributions. By using the Principal Component Analysis (PCA) and FastICA algorithm we determined the number of components, and defined the statistically independent components (ICs) in the fractal complexity of signal waveforms. We have isolated two independent components of the empirical FD distributions for each of three groups of data by using FastICA algorithm. ICs represent the sources of fractal waveforms complexity of Br neuron activity in particular experimental conditions. Our main results have shown that there could be two opposite intrinsic mechanisms in single snail Br neuron response to static magnetic field stimulation. We named identified ICs that correspond to those mechanisms - the component of plasticity and the component of elasticity. We have shown that combination of fractal analysis with ICA method could be very useful for the decomposition and identification of the sources of fractal complexity of bursting neuronal activity waveforms.

Cortico-pontine theta synchronization phase shift following monoaminergic lesion in rat.

The experiments were performed in 14 adult, male Sprague Dawley rats chronically instrumented for sleep recording and recorded during baseline condition, following sham injection (saline i.p. 1 ml/kg), and every week for 5 weeks following injection of the systemic neurotoxins (DSP-4 or PCA; 1 ml/kg, i.p.) for chemical axotomy of the locus coeruleus (LC) and dorsal raphe (DR) axon terminals. In our former study we demonstrated that the systemically induced lesion of the noradrenergic or serotonergic axon terminals did not affect the sleep-wake distribution from control condition. In this study, by using spectral analysis and phase shift spectra of the cortical and pontine EEG we analyzed cortico-pontine theta oscillation synchronization phase shift on 6-hour recordings in control condition and 28 days following the monoaminergic lesions, as a time for permanently established DR or LC chemical axotomy. Our results demonstrated for the first time that chronically decreased brain monoamines in freely moving rats changed cortico-pontine theta synchronization phase shift. Pons became a leading theta oscillator. We assume that deficit of monoamines induced predominance of the NREM/REM transitions, characterized with phasic theta oscillations (the increased density of clustered P waves which intrinsic frequency corresponds to theta frequency oscillations), and may produced preceding phasic theta versus tonic theta oscillation drive.

Monoaminergic system lesions increase post-sigh respiratory pattern disturbance during sleep in rats.

Monoamines are important regulators of behavioral state and respiratory pattern, and the impact of monoaminergic control during sleep is of particular interest for the stability of breathing regulation. The aim of this study was to test the effects of systemically induced chemical lesions to noradrenergic and serotonergic efferent systems, on the expression of sleep-wake states, pontine wave activity, and sleep-related respiratory pattern and its variability. In chronically instrumented male adult Sprague-Dawley rats we lesioned noradrenergic terminal axonal branches by a single intraperitoneal dose of DSP-4 (N-(2-chloroethyl)-N-ethyl-2-brombenzilamine; 50 mg/kg, i.p.), and serotonergic axonal terminals by two intraperitoneal doses, 24 h apart, of PCA (p-chloroamphetamine; 6 mg/kg, i.p.). In each animal, we recorded sleep, pontine waves (P-waves) and breathing at baseline, following sham injection, and every week for 5 weeks following injection of either systemic neurotoxin. Distinct responses were observed to the two lesions. DSP-4 lesions were associated with a trend toward increased NREM sleep (p < 0.06), decreased wakefulness (p < 0.05) and increased respiratory tidal volume during NREM (p = 0.0002) and REM (p = 0.0001) sleep with respect to baseline. None of these effects, however, were observed during the first 14 days after injection. No significant changes were observed in the frequency of apneas or sighs, nor in the coupling between these two, at any time after DSP-4 injection. Conversely, selective serotonergic lesion by PCA produced no change in the baseline respiratory frequency or tidal volume during sleep or wakefulness, nor was the expression of Wake, NREM or REM sleep affected. Instead, PCA injection resulted in a sustained increase in the frequency and duration of post-sigh apneas (PS) during NREM sleep (p = 0.002). This reflected increased coupling between sighs and apneas, because neither the frequency nor the amplitude of spontaneous sighs was altered by PCA.

C-fiber activation exacerbates sleep-disordered breathing in rats.

We reported previously that activation of vagal feedback by protoveratrines or serotonin exacerbates sleep apnea in rats, but each of these agents activates multiple afferent fiber types. To elucidate the specific impact of C-fiber activity on sleep apnea, the present study utilized capsaicin (CAP), which stimulates C-fibers via the CAP receptor. Nine adult Sprague-Dawley rats were instrumented for chronic polysomnography and recorded for 6 hours on four occasions. Prior to each recording, the animals received an intraperitoneal injection of either saline (control), or CAP 0.1, 1.0, or 10.0 mg/kg. Respiration was monitored by single-chamber plethysmography and apneas were scored as breaths longer than 2.5 seconds not preceded by a sigh. CAP increased apneas during non-rapid eye movement (NREM) sleep (p < 0.05 vs control) and reduced respiratory minute ventilation by about 15% in all behavioral states (waking, NREM, and REM sleep). We conclude that selective pharmacological activation of C-fibers produces a diathesis of sleep-disordered breathing specific to NREM sleep in rats.

Interspike background activity in extracelullarly recorded Purkinje neurons: spectral analysis.

The aim of this study was to investigate the spectral characteristics of Purkinje cell interspike background activity caused by the occurrence of particular action potentials or by electrically induced enhancement of cerebellar inhibitory and excitatory input drive. Spontaneously active Purkinje neurons were extracellularly recorded in anesthetized rats before and after cessation of stimulation from the inferior olive (10) or locus coeruleus (LC). After A/D conversion (30 kHz), direct spectral analysis of extracted interspike background activity was done. Our results have shown that, in contrast to simple spikes, the occurrence of complex spikes induces changes in the spectra of interspike background activity. The different spectral changes of interspike background activity induced by LC and 10 stimulation also indicated the importance of this extracellularly recorded phenomenon.

Effect of early cortical lesion on the acute model of epilepsy.

The experiments were performed in order to investigate the sparing of function following early postnatal cortical lesion in the acute rat model of epilepsy. Sensorimotor cortex was unilaterally removed at 9 and 10 days of postnatal age in lesioned animals, while control animals were only sham operated (at the same early stage of life) or non-operated (before implantation of the electrodes). Seizure activity was recorded by means of electroencephalograms at adult stage of life induced by parenteral administration of penicillin (1,000,000 I.U./kg, i.p.). Our results showed that when the cortical lesion was performed in infancy (on the contrary to the lesion performed in adulthood) there was no prolongation of seizure activity in an acute model of epilepsy.

Effect of neurotoxin DSP4 on EEG power spectra in the rat acute model of epilepsy.

The effect of the adrenergic neurotoxin N-(chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4) on electroencephalographic (EEG) activity was studied in the model of epilepsy induced by systemic application of penicillin (1,000,000 IU/kg, i.p). DSP4 (50 mg/kg, i.p.) was administrated to male Wistar rats, while the control animals were rats from the same litters. EEG activity was recorded in acute and chronic experiments 3 or 4 weeks after DSP4 treatment, before and after penicillin administration. Occasional locus coeruleus (LC) stimulation served as an electrophysiological test of DSP4 toxic effect. EEG power spectra in DSP4 treated animals showed a tendency to be greater in lower frequency bands than in controls before penicillin administration; there was almost no effect of electrical LC stimulation, regardless on penicillin treatment. In the model of epilepsy, the mean total EEG power spectra were greater in the period of 135-330 min after penicillin administration, as well as during 345-540 min, in DSP4 treated animals as compared to the controls. It seems that neurotoxin DSP4 is an optimal tool for studying the removal of LC influence in the acute model of epilepsy. It is also suggested that norepinephrine (NE) may have a modulatory role in the systemic penicillin epilepsy.

Changes in neuropeptide levels after brain damage in rats.

The physiological and pathophysiological roles of neuropeptides are still not clear. The aim of our study was to detect long lasting changes of vasoactive-intestinal peptide (VIP), somatostatin (SOM) and substance P (SP) contents in the rat cerebral cortex and hippocampus after brain lesion. The experiments were performed on groups of adult male Wistar rats. The first group consisted of animals with unilateral ablation of the sensorimotor cortex performed at the age of 60 days. The second group was a control one (rats of the same age but with an intact brain). Both groups of animals were sacrificed at the age of 90-105 days and radioimmunoassay was used to determine amounts of VIP, SOM and SP. The mean values of VIP levels were decreased significantly only in contralateral cortical areas, while there was an increase of SP in lesioned animals. Our results suggest that descrete changes in neuropeptide levels occur during restorative processes after brain lesion.

Amphetamine and haloperidol modulatory effects on Purkinje cell activity and on EEG power spectra in the acute rat model of epilepsy.

The modulation of cerebellar Purkinje cell activity and EEG from parietal cortex was studied in the rat model of epilepsy induced by penicillin under acute haloperidol and amphetamine treatment. The discharge pattern of Purkinje cells showed tendency towards inhibition and EEG power spectra increased after parenteral administration of penicillin (1000000 IU/kg, i.p.). Acute haloperidol treatment (1 mg/kg, i.p.), performed after the development of penicillin induced epileptic episodes, elicited a prominent excitation of Purkinje cell discharges associated with parallel increase in mean EEG power spectra. However, acute DL-amphetamine treatment induced marked suppression of Purkinje cell discharges as well as outstanding decrease of the mean EEG power spectra. These results indicate that cerebellar Purkinje cells may be important in the control of seizure activity and that noradrenergic influences are relevant.

Effect of cerebellar stimulation on EEG power spectra in the acute model of epilepsy.

Experiments were performed to investigate the effects of cerebellar stimulation on epilepsy induced by parenteral administration of penicillin, in rats without or with the lesion of sensorimotor cortex. There were no differences in the EEG activity of the same experimental animal after the first and subsequent penicillin treatments (at least 7 days later). The electrical stimulation (duration of 5-10 min) of the lateral cerebellar nucleus was applied repetitively 135-315 min after penicillin administration, when the EEG power spectra markedly increased. The cerebellar stimulation evoked the decrease of the mean total EEG power spectra, but the effects were temporary. The EEG power spectra were significantly lower (P < 0.05) during the period of 150-330 min after penicillin treatment in experimental sessions with applied cerebellar stimulation in comparison to the experimental sessions without such stimulation. The residual effects (if any) of cerebellar stimulation on the EEG activity in the later period, 345-600 after penicillin treatment were not significant (P > 0.05). Cerebellar stimulation had the same effect among unlesioned animals and animals with prior cortical lesion in the acute model of epilepsy.

The effect of cortical lesion on systemic penicillin epilepsy in rats.

There is a certain recovery of function following brain damage, due to neuronal plasticity. The experiments were performed in order to investigate the effects of cortical lesion on seizural activity in rats induced by systemic application of penicillin. The sensorimotor cortex was unilaterally removed in the lesioned animals, while the control animals were only sham operated or non-operated (before implantation of the electrodes). Seizural activity was recorded by means of electroencephalograms before and after penicillin treatment (1,000,000 I.U./kg, i.p). Testing of penicillin started at least 30 days after cortical lesion. Seizural activity was characterized by spike and wave complexes accompanied by vigilance reduction and sometimes by mild myoclonic jerks in both control and lesioned animals. The early period (about 2 h after penicillin administration) with appearance of the spike-wave discharges with relative increase of the mean total electroencephalogram powers as well as the succeeding period 2.5-5.5 h after penicillin administration) with maximum number of spike-wave discharges did not differ in the electroencephalogram of the control and lesioned animals. The late period of penicillin effect (from 6-11 h after penicillin administration) with frequent spike-wave discharges and still large mean total electroencephalogram powers was observed only in lesioned animals. It is concluded that a cortical lesion destabilizes the brain function in the rat model of epilepsy induced by parenteral administration of penicillin.

Activity of cat cerebellar neurons in penicillin epilepsy and amphetamine treatment.

The discharge pattern of cerebellar Purkinje cells and fastigial neurons was studied after acute amphetamine treatment in immobilized cats, as well as during generalized penicillin-induced epilepsy. There was a marked reversible decrease in spontaneous firing rate of Purkinje cells and an increase in spontaneous firing rate of fastigial neurons after acute d-1 amphetamine administration (5 mg/kg, s.c.). The discharge pattern of Purkinje cells showed tendency towards inhibition, while the fastigial neurons showed less clear tendency towards disinhibition in the course of epilepsy induced by parenteral administration of penicillin (400.000-500.000 I.U./kg, i.m.). Moreover, acute amphetamine treatment (5 mg/kg, s.c.) performed after the development of penicillin-induced epileptic episodes elicited a prominent suppression of Purkinje cell discharges associated with a parallel increase in discharges of fastigial neurons. These results suggest that the changes in discharge rate of cerebellar corticonuclear neurons induced by amphetamine contribute to suppression of seizural activity in the feline model of generalized epilepsy.