Unanesthetized Cats Research Articles

0066 Glycinergic Postsynaptic Inhibition is Responsible for the Suppression of Hypoglossal Motoneuron Activity During Naturally-Occurring REM Sleep

Abstract Introduction The present study was undertaken to explore the role of glycinergic postsynaptic inhibition and monoaminergic disfacilitation (a withdrawal of excitatory noradrenergic and serotonergic inputs) in the control of hypoglossal motoneuron activity during REM sleep. Accordingly, glycinergic, noradrenergic and serotonergic antagonists were microinjected into the hypoglossal nucleus, and their effects on the hypoglossal nerve activity during REM sleep were examined in chronically-instrumented, unanesthetized cats. Methods Adults cats were prepared for monitoring behavioral states of sleep and wakefulness, and for extracellular recordings from hypoglossal nerve. Strychnine (a glycinergic antagonist) and a mixture of prazosin (a noradrenergic antagonist) and methysergide (a serotonergic antagonist) were microinjected, separately, into the hypoglossal nucleus during naturally-occurring states of sleep and wakefulness. Results During REM sleep, compared to non-REM sleep, the hypoglossal nerve activity decreased by 17.4±1.5% (n=17) in the control recordings (prior to the injection of strychnine). Following the microinjection of strychnine, there was only a mean decrease of 7.2±1.2% (n=12) in the nerve activity during REM sleep versus NREM sleep. The strychnine effect was statistically significant compared to control (p<0.001; unpaired t-test), which indicates that strychnine blocks REM sleep-related suppression of hypoglossal nerve activity. In contrast, the microinjection of prazosin and methysergide did not significantly reduce the hypoglossal nerve activity during REM sleep (control: 15.9±2.3, n=9 vs. prazosin+methysergide: 12.6±1.4%, n=10, p=0.229, unpaired t-test). Conclusion The present results demonstrate that the microapplication of strychnine, but not prazosin and methysergide, into the hypoglossal nucleus significantly reduces the suppression of the hypoglossal nerve activity during naturally-occurring REM sleep. We therefore suggest that glycinergic postsynaptic inhibition is primarily responsible for the suppression of hypoglossal motoneuron activity during REM sleep. Support 5R01NS094062

Blockade of B2 receptors attenuates the responses of group III afferents to static contraction

Recent evidence has been presented demonstrating that group III mechanoreceptors comprise an important part of the sensory arm of the exercise pressor reflex, which in turn functions to increase arterial blood flow to contracting skeletal muscles. Although group III afferents are stimulated by mechanical distortion of their receptive fields, they are also stimulated by bradykinin, which is produced by skeletal muscle when it contracts. Moreover, blockade of B (bradykinin)2 receptors has been shown to decrease the magnitude of the exercise pressor reflex. Nevertheless, the effect of blockade of B2 receptors on responses of group III afferents to contraction is not known. We therefore determined the effect of B2 receptor blockade with HOE 140 (40μg/kg) on the responses to both static and intermittent contraction of group III afferents with endings in the triceps surae muscle of decerebrated unanesthetized cats. We found that HOE 140 significantly attenuated (P=0.04) the responses of 14 group III afferents to static contraction, but did not significantly attenuate (P=0.16) the responses of 16 group III afferents to intermittent contraction. The attenuation induced by HOE 140 was present throughout the static contraction period, and led us to speculate that blockade of B2 receptors on the endings of group III afferents decreased their sensitivity to mechanical events occurring in the working muscles.

Effect of Microiontophoretic Application of Modulators of Noradrenergic Transmission on Impulse Activity of Neurons of the Cat Motor Cortex during Performance of the Reflex to a Complex of Stimuli

On unanesthetized cats, we studied the effects of selective modulators of noradrenergic transmission on the activity of neurons of the cerebral motor cortex in the course of realization of an operant motor reflex to presentation of a complex of stimuli. These modulators were applied using microiontophoretic injections into sites of recording of impulse activity of cortical neurons within the zone of projection of the “working” forelimb. Applications of the α1 receptor agonist Mesaton resulted in significant suppression of background impulsation of the neurons and spiking within the interstimulus interval immediately during realization of the movement. Under the action of the α2 blocker yohimbine, opposite effects were observed. The activity of neurons increased within both background period and other examined time intervals. Mechanisms of the effects of modulation of noradrenergic transmission in the sensorimotor cortex are discussed.

Coordination of cough and swallow in vivo and in silico

The respiratory pattern generation network of the ventrolateral medulla reconfigures to produce cough and swallow. The only extant network model of breathing, cough and swallow is based on data from decerebrated, unanesthetized cats. We compared in silico simulations to in vivo data from anesthetized cats. Coughs were elicited by mechanical stimulation of the trachea. Electromyograms (EMG) of the parasternal (PS) and transversus abdominis (TA) muscles identified cough. Swallow was elicited by injection of water into the pharynx and EMGs of the thyroarytenoid (ThAr), PS, and geniohyoid (GH) muscles identified swallow. Simulations predicted PS EMG amplitude of “post-swallow” cough would be greater than “pre-swallow” cough and this effect was observed in vivo. Simulations also reproduced in vivo finding of greater ThAr EMG magnitude during swallow than cough. However, important temporal features that govern cough and swallow coordination when both behaviors are co-expressed were not observed in silico, such as, during repetitive coughing, the expiratory phase (E2) of cough is prolonged when swallow is present. We conclude the current model is predictive; the in silico simulations reproduce several important features of in vivo airway protection; however some temporal relationships between cough and swallow are not currently accounted for by the model. Supported by NIH HL 89104; HL 89071; HL 103415.

Midbrain and medullary control of postinspiratory activity of the crural and costal diaphragm in vivo

Studies on brain stem respiratory neurons suggest that eupnea consists of three phases: inspiration, postinspiration, and expiration. However, it is not well understood how postinspiration is organized in the diaphragm, i.e., whether postinspiration differs in the crural and costal segments of the diaphragm and what the influence is of postinspiratory neurons on diaphragm function during eupnea. In this in vivo study we investigated the postinspiratory activity of the two diaphragm segments during eupnea and the changes in diaphragm function following modulation of eupnea. Postinspiratory neurons in the medulla were stereotaxically localized extracellularly and neurochemically stimulated. We used three types of preparations: precollicularly decerebrated unanesthetized cats and rats and anesthetized rats. In all preparations, during eupnea, postinspiratory activity was found in the crural but not in the costal diaphragm. When eupnea was discontinued in decerebrate cats in which stimulation in the nucleus retroambiguus induced activation of laryngeal or abdominal muscles, all postinspiratory activity in the crural diaphragm was abolished. In decerebrate rats, stimulation of the midbrain periaqueductal gray abolished postinspiration in the crural diaphragm but induced activation in the costal diaphragm. In anesthetized rats, stimulation of medullary postinspiratory neurons abolished the postinspiratory activity of the crural diaphragm. Vagal nerve stimulation in these rats increased the intensity of postinspiratory neuronal discharge in the solitary nucleus, leading to decreased activity of the crural diaphragm. These data demonstrate that three-phase breathing in the crural diaphragm during eupnea exists in vivo and that postinspiratory neurons have an inhibitory effect on crural diaphragm function.

Effects of Local and Remote Muscle Pain on Stretch ReflexActivities in the Elbow Joint Flexors and Extensors of Unanesthetized Cats

In experiments on unanesthetized cats, we compared the effects of experimentally induced pain in the m. biceps brachii or in the neck muscles on EMG activity of the flexors and extensors of the elbow joint (mm. biceps et triceps brachii, respectively) evoked by a passive extension-flexion of the above joint. Muscle pain was induced by injections of 0.5 ml of a hypertonic (7%) NaCl solution into the above-mentioned muscles. In the case of pain in the biceps, i.e., in the muscle directly involved in realization of the reflex, we observed an increase in the amplitude and significant shortening of the latency of EMG responses of this muscle. The amplitude of a short-latency (supposedly monosynaptic) component of the biceps reflex (М1 response) increased by 65%, while an increment of the latter (supposedly polysynaptic) М2 component was 117%. When pain was induced in anatomically remote neck muscles, the stretch reflex in the biceps was considerably suppressed. The maximum amplitudes of the М1 and М2 components decreased by 25 and 30%, respectively, but the latencies of these components decreased significantly, similarly to what was observed in the case of induction of experimental pain in the biceps. Under both conditions of experimental pain, changes in the parameters of EMG responses of the forearm extensor (m. triceps brachii) demonstrated similarity with those of the biceps responses. The maximum effect of pain induction was observed within the first 5 min after injections of the hypertonic solution; full recovery of the stretch reflex parameters was observed on the 20th to 30th min. We conclude that the effects of pain induction on the reflex under study are not generalized. They depend on the site of such induction with respect to the muscle where the stretch reflex is elicited. Unidirectional effects of both types of pain on the antagonist muscles allow us to suppose that modulation of the reflex reactions upon pain induction is mediated by influences from the supraspinal CNS structures. Induction of pain in the biceps increased the amplitude of EMG manifestations of the stretch reflex, while such induction in the neck muscles decreased such responses; nonetheless, in both cases the latency of the reflexes decreased. This fact allows us to believe that the sensitivity of muscle spindles increased under both conditions of the pain influence.

Dr Werner P Koella, 1917-2008

Dr Werner P Koella, aged 91 years, passed away on 13 January 2008. After graduating from the University of Zurich in 1942, he started his career at the Clinic of Neurosurgery of the Zurich University Hospital. After 2 years of clinical activity, his interest in the functioning of the central nervous system led him to join the Institute of Physiology. Under the guidance of Professor WR Hess, Nobel laureate in medicine and physiology, he first directed his research efforts toward the experimental study of nystagmus and the role of the hypothalamus in fluid excretion. He made use of the technique of electrical brain stimulation in the unanesthetized cat and reported in 1952 together with K Akert and R Hess Jr on the sleep-inducing action of thalamic stimuli. In the following year, the same authors published the paper ‘Cortical and subcortical recordings in natural and artificially induced sleep in cats,’ which has been highly cited and marks the beginning of Koella's career in experimental sleep research. In 1951, he moved to the Department of Physiology of the University of Minnesota, where he joined the neurophysiology laboratory of Ernst Gellhorn. There he studied the effects of ambient temperature on the EEG and began to use the method of evoked responses to monitor central nervous structures. From Minneapolis he moved to Shrewsbury, Massachusetts, where he joined Peter J Morgane at the Worcester Foundation of Experimental Biology. From very early on, Werner Koella recognized the important role of serotonin in brain mechanisms. Already in 1959, he published the first papers on the effects of serotonin and LSD on evoked potentials. In the following years, serotonin became the major focus of his research, culminating in his most highly cited paper on the effect of serotonin depletion by parachlorophenylalanine on sleep, which he published in 1968 with Feldstein and Czicman. Because at that time serotonin was considered to be the major sleep-promoting transmitter, he suggested jocularly that its name be changed to somnotonin.

Effect of Hypercapnia on Sleep and Breathing in Unanesthetized Cats

In this study, we looked at the effect of hypercapnia on sleep architecture and breathing. We characterized the effect of hypercapnia on duration, frequency, and latency of NREM and REM sleep. We described state-specific patterns of breathing as well. This study is relevant to understand possible treatments for sleep disordered breathing. Four cats were studied during 3-hour sessions while breathing 0%, 2%, 4%, and 6% CO2 in room air. Each animal was studied 4 days per week for a period of 4 weeks. The animals breathed through a tube inserted into the trachea via a surgically created fistula. Respiration was measured using pneumotachography, and brain activity was recorded from implanted electrodes to discriminate states of sleep and wakefulness. Two percent inspired CO2 increased sleep duration and decreased time awake. On the other hand, 6% CO2 induced a worsening of sleep parameters: the duration of wakefulness increased by 24.2%. As a response to hypercapnia, tidal volume (V(T)), minute ventilation (V(E)), and respiratory effort (V(T)/T(I)) increased proportionally in all states with increasing levels of CO2. With 6% CO2, breathing tended to become similar in all states of consciousness. All breathing parameters converged towards a common value independently of the states. We conclude that a mild hypercapnic stimulus can stimulate both breathing and sleep, and it may be useful in treatment of sleep disordered breathing.

Role played by P2X and P2Y receptors in evoking the muscle chemoreflex

The role played by purinergic 2Y receptors in evoking the muscle chemoreflex is not well defined. To shed light on this issue, we compared the pressor responses with popliteal arterial injection of UTP (1 mg/kg), a selective P2Y agonist, with those to popliteal arterial injection of ATP (1 mg/kg), a P2X and P2Y agonist, and to alpha,beta-methylene ATP (50 mug/kg), a selective P2X1 and P2X3 agonist, in decerebrate unanesthetized cats. We found that injection of ATP and alpha,beta-methylene ATP increased mean arterial pressure by 19 +/- 2 and 15 +/- 4 mmHg, whereas UTP had no affect on arterial pressure. In addition, the pressor responses to injection of ATP and alpha,beta-methylene ATP were abolished by section of the sciatic nerve, demonstrating that they were reflex in origin. We conclude that P2Y receptors on thin fiber muscle afferents play no role in evoking the muscle chemoreflex.

Effects of modulation of GABA-and noradrenergic transmission on impulse activity of neurons of the cat motor cortex related to realization of an operant reflex to the action of two signals

On unanesthetized cats trained to perform placing movements to the action of two subsequent signals (warning and imperative stimuli), we examined reflex-related impulse activity (IA) of neurons of the motor cortex (field 4) and simultaneous changes in the “slow” cortical potentials (SCP) in the vertex zone. In almost all cases under study, the shift in the SCP toward negativity was associated with a decrease in the frequency of IA within interstimulus intervals; this corresponded to a period of focusing of the animal’s attention on the expected imperative stimulus. Using a microiontophoretic technique, we tried to elucidate the role of GABA-and adrenergic cerebral systems in the genesis of such inhibitory periods. We conclude that, independently of each other but synchronously, both these systems can be involved in the maintenance of processes of active inhibition in the cerebral cortex under conditions of realization of an operant reflex.

Dorsal cochlear nucleus response properties following acoustic trauma: Response maps and spontaneous activity

Recordings from single neurons in the dorsal cochlear nucleus (DCN) of unanesthetized (decerebrate) cats were done to characterize the effects of acoustic trauma. Trauma was produced by a 250 Hz band of noise centered at 10 kHz, presented at 105–120 dB SPL for 4 h. After a one-month recovery period, neurons were recorded in the DCN. The threshold shift, determined from compound action-potential audiograms, showed a sharp threshold elevation of about 60 dB at BFs above an edge frequency of 5–10 kHz. The response maps of neurons with best frequencies (BFs) above the edge did not show the typical organization of excitatory and inhibitory areas seen in the DCN of unexposed animals. Instead, neurons showed no response to sound, weak responses that were hard to tune and characterize, or “tail” responses, consisting of broadly-tuned, predominantly excitatory responses, with a roughly low-pass shape similar to the tuning curves of auditory nerve fibers with similar threshold shifts. In some tail responses whose BFs were near the edge of the threshold elevation, a second weak high-frequency response was seen that suggests convergence of auditory nerve inputs with widely separated BFs on these cells. Spontaneous rates among neurons with elevated thresholds were not increased over those in populations of principal neurons in unexposed animals.

Neuronal mechanisms of active (rapid eye movement) sleep induced by microinjections of hypocretin into the nucleus pontis oralis of the cat

Hypocretinergic (orexinergic) neurons in the hypothalamus project to the nucleus pontis oralis, a nucleus which plays a crucial role in the generation of active (rapid eye movement) sleep. We recently reported that the microinjection of hypocretin into the nucleus pontis oralis of chronically-instrumented, unanesthetized cats induces a behavioral state that is comparable to naturally-occurring active sleep. The present study examined the intracellular signaling pathways underlying the active sleep-inducing effects of hypocretin. Accordingly, hypocretin-1, a protein kinase C inhibitor and a protein kinase A inhibitor were injected into the nucleus pontis oralis in selected combinations in order to determine their effects on sleep and waking states of chronically instrumented, unanesthetized cats. Microinjections of hypocretin-1 into the nucleus pontis oralis elicited active sleep with a short latency. However, a pre-injection of bisindolylmaleimide-I, a protein kinase C-specific inhibitor, completely blocked the active sleep-inducing effects of hypocretin-1. The combined injection of bisindolylmaleimide-I and hypocretin-1 significantly increased the latency to active sleep induced by hypocretin-1; it also abolished the increase in the time spent in active sleep induced by hypocretin-1. On the other hand, the injection of 2′5′-dideoxyadenosine, an adenylyl cyclase inhibitor, did not block the occurrence of active sleep by hypocretin-1. We conclude that the active sleep-inducing effect of hypocretin in the nucleus pontis oralis is mediated by intracellular signaling pathways that act via G-protein stimulation of protein kinase C.

Barosensory cells in the nucleus tractus solitarius receive convergent input from group III muscle afferents and central command

Some neural mechanism must prevent the full expression of the baroreceptor reflex during static exercise because arterial blood pressure increases even though the baroreceptors are functioning. Two likely candidates are central command and input from the thin fiber muscle afferents evoking the exercise pressor reflex. Recently, activation of the mesencephalic locomotor region, an anatomical locus for central command, was found to inhibit the discharge of nucleus tractus solitarius cells that were stimulated by arterial baroreceptors in decerebrated cats. In contrast, the effect of thin fiber muscle afferent input on the discharge of nucleus tractus solitarius cells stimulated by baroreceptors is not known. Consequently in decerebrated unanesthetized cats, we examined the responses of barosensory nucleus tractus solitarius cells to stimulation of thin fiber muscle afferents and to stimulation of the mesencephalic locomotor region, a maneuver which evoked fictive locomotion. We found that electrical stimulation of either the mesencephalic locomotor region or the gastrocnemius nerve at current intensities that recruited group III afferents inhibited the discharge of nucleus tractus solitarius cells receiving baroreceptor input. We also found that the inhibitory effects of both gastrocnemius nerve stimulation and mesencephalic locomotor region stimulation converged onto the same barosensory nucleus tractus solitarius cells. We conclude that the nucleus tractus solitarius is probably the site whereby input from both central command and thin fiber muscle afferents function to reset the baroreceptor reflex during exercise.

Medullary Respiratory Neural Activity During Hypoxia in NREM and REM Sleep in the Cat

Intact unanesthetized cats hyperventilate in response to hypocapnic hypoxia in both wakefulness and sleep. This hyperventilation is caused by increases in diaphragmatic activity during inspiration and expiration. In this study, we recorded 120 medullary respiratory neurons during sleep in hypoxia. Our goal was to understand how these neurons change their activity to increase breathing efforts and frequency in response to hypoxia. We found that the response of medullary respiratory neurons to hypoxia was variable. While the activity of a small majority of inspiratory (58%) and expiratory (56%) neurons was increased in response to hypoxia, the activity of a small majority of preinspiratory (57%) neurons was decreased. Cells that were more active in hypoxia had discharge rates that averaged 183% (inspiratory decrementing), 154% (inspiratory augmenting), 155% (inspiratory), 230% (expiratory decrementing), 191% (expiratory augmenting), and 136% (expiratory) of the rates in normoxia. The response to hypoxia was similar in non-rapid-eye-movement (NREM) and REM sleep. Additionally, changes in the profile of activity were observed in all cell types examined. These changes included advanced, prolonged, and abbreviated patterns of activity in response to hypoxia; for example, some inspiratory neurons prolonged their discharge into expiration during the postinspiratory period in hypoxia but not in normoxia. Although changes in activity of the inspiratory neurons could account for the increased breathing efforts and activity of the diaphragm observed during hypoxia, the mechanisms responsible for the change in respiratory rate were not revealed by our data.

Cortical modulation of neuronal activity in the cat's lateral geniculate and perigeniculate nuclei

The cortico-thalamic influence on spontaneous and visually evoked activity of single cells in the dorsal lateral geniculate (LGN) and perigeniculate (PGN) nuclei were examined in unanesthetized cats with pretrigeminal brainstem transections by means of reversible cooling of cortical areas 17 and 18. The spatio-temporal characteristic of cells' RFs was tested with light spot randomly presented at different points along the receptive field axis. The cessation of cortical input decreased spontaneous activity of most of the LGN cells (64%; as compared to 36% with increased background firing). Similarly, their visually evoked responses were reduced (70% cells; compared to 24% with increased response) and extent of central excitatory domains diminished. In contrast, the majority of PGN neurons increased their spontaneous activity (62%; compared to 38% with decreased firing rate). Cortical cooling resulted also in a decrease of the ON and OFF central responses of most PGN cells (55%; as compared to 20% with increased responses). The described effects were more pronounced within the population of cells in X than in Y pathway. Although the removal of descending cortical excitation disturbed the balance of activity within the network of thalamic cells the gain of the geniculate relay was preserved. We conclude that the main role exerted by the cortico-thalamic pathway serves facilitation of the ascending retino-cortical flow of visual information at the level of lateral geniculate nucleus.

Interactions between GABAergic and Cholinergic Processes in the Nucleus Pontis Oralis: Neuronal Mechanisms Controlling Active (Rapid Eye Movement) Sleep and Wakefulness

The cholinergic system within the nucleus pontis oralis (NPO) of the pontine tegmentum is critically involved in the generation of active (rapid eye movement) sleep. Previously, we demonstrated that a GABAergic system in the NPO also plays an important role in the control of the behavioral states of wakefulness as well as active sleep. The present study examined interactions between these two neuronal systems vis-a-vis the occurrence of these behavioral states. Accordingly, cholinergic and GABAergic agonists and antagonists were injected into the NPO, and their combined effects on sleep and waking states of chronic, unanesthetized cats were examined. Microinjections of carbachol into the NPO elicited active sleep with a short latency. However, a preinjection of muscimol (a GABA(A) agonist) completely blocked the active sleep-inducing effects of carbachol. The induction of active sleep by carbachol was also suppressed by a subsequent injection of muscimol. On the other hand, the microinjection of scopolamine (a muscarinic receptor antagonist) did not block the induction of active sleep by bicuculline (a GABA(A) antagonist). We conclude that the excitatory cholinergic control of NPO neurons that are involved in the generation of active sleep is gated by a pontine GABAergic system that exerts its effects postsynaptically by inhibiting NPO neurons, resulting in the suppression of active sleep and the generation of wakefulness. In the absence of the activation of this GABAergic gating mechanism, active sleep occurs. These results reveal that specific interactions between cholinergic and GABAergic processes in the NPO play a critical role in the generation of active sleep and wakefulness.

Evoked Potentials of the Somatic Cortex and EMG Reactions of the Shoulder Muscles Elicited by Passive Extension of the Elbow Joint in Unanesthetized Cats

We recorded electromyographic (EMG) reactions from the flexors of the elbow joint and evoked potentials (EP) from the somatic cortex (fields 3, 4, and 6) of unanesthetized cats. These reactions were elicited by perturbation of an external extensor loading applied to the arm and evoking passive extension of the elbow joint. Perturbation of the loading was performed in two modes: (i) with different fixed force moments within a 0.04–0.2 N·m range, but with a constant rate of change in this moment (3.2 N·m·sec−1), and (ii) with a constant force moment magnitude (0.2 N·m), but with different rates of change in this moment (from 0.1 to 6.4 N·m·sec−1). When the elbow joint was passively extended, an EMG response was generated in the m. biceps brachii. The amplitude of this response correlated with the amplitude of perturbation of the external loading, and the time course of the response was rather close to that of the evoked passive moment. It was possible to differentiate several (up to seven) successive components in EP recorded from the three above-mentioned cortical fields; among them, the component N(50–60) was the most stable and clearly manifested. Its amplitude did not depend on the level of external loading and decreased with a decrease in the rate of loading perturbation. The time course of the N(50–60) changed insignificantly with variation of temporal parameters of the stimulus and of the evoked movement. We conclude that the spinal level and the cortical level responsible for formation of the stretch reflex differ significantly from each other in their functional roles. Reactions of the spinal level (which could be characterized by changes in EMG) are to a greater extent related to a change in the position of the limb link, while reactions of the cortical level (EP) are determined by the arrival of information about changes in the forces applied to the joint. Neurons of the somatic cortex, which are excited in the course of the stretch reflex, cannot be considered the main source responsible for generation of the M2 component of the myographic response. It is supposed that the cortical level predetermines the formation of non-reflex motor commands related to motor reflexes closed in the somatic brain cortex.

Alpha,beta-Methylene ATP elicits a reflex pressor response arising from muscle in decerebrate cats.

In part, the exercise pressor reflex is believed to be evoked by chemical stimuli signaling that blood supply to exercising muscles is not adequate to meet its metabolic demands. There is evidence that either ATP or adenosine may function as one of these chemical stimuli. For example, muscle interstitial concentrations of both substances have been found to increase during exercise. This finding led us to test the hypothesis that popliteal arterial injection of alpha,beta-methylene ATP (5, 20, and 50 microg/kg), which stimulates P2X receptors, and 2-chloroadenosine (25 microg/kg), which stimulates P1 receptors, evokes reflex pressor responses in decerebrate, unanesthetized cats. We found that popliteal arterial injection of the two highest doses of alpha,beta-methylene ATP evoked pressor responses, whereas popliteal arterial injection of 2-chloroadenosine did not. In addition, the pressor responses evoked by alpha,beta-methylene ATP were blocked either by section of the sciatic nerve or by prior popliteal arterial injection of pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid (10 mg/kg), a selective P2-receptor antagonist. We conclude that the stimulation of P2 receptors, which are accessible through the vascular supply of skeletal muscle, evokes reflex pressor responses. In addition, our findings are consistent with the hypothesis that the stimulation of P2 receptors comprises part of the metabolic error signal evoking the exercise pressor reflex.

Induction of active (REM) sleep and motor inhibition by hypocretin in the nucleus pontis oralis of the cat.

Hypocretin (orexin)-containing neurons in the hypothalamus, which have been implicated in the pathology of narcolepsy, project to nuclei in the brain stem reticular formation that are involved in the control of the behavioral states of sleep and wakefulness. Among these nuclei is the nucleus pontis oralis (NPO). Consequently, the present study was undertaken to determine if the hypocretinergic system provides regulatory input to neurons in the NPO with respect to the generation of the states of sleep and wakefulness. Accordingly, polygraphic recordings and behavioral observations were obtained before and after hypocretin-1 and -2 were microinjected into the NPO in chronic, unanesthetized cats. Microinjections of either hypocretin-1 or -2 elicited, with a short latency, a state of active [rapid eye movement (REM)] sleep that appeared identical to naturally occurring active sleep. The percentage of time spent in active sleep was significantly increased. Dissociated states, which are characterized by the presence of muscle atonia without one or more of the electrophysiological correlates of active sleep, also arose following the injection. The effect of juxtacellular application of hypocretin-1 on the electrical activity of intracellularly recorded NPO neurons was then examined in the anesthetized cat. In this preparation, the application of hypocretin-1 resulted in the depolarization of NPO neurons, an increase in the frequency of their discharge and an increase in their excitability. These latter data represent the first description of the in vivo action of hypocretin on intracellularly recorded neuronal activity and provide evidence that the active sleep-inducing effects of hypocretin are due to a direct excitatory action on NPO neurons. Therefore we suggest that hypocretinergic processes in the NPO may play a role in the generation of active sleep, particularly muscle atonia and therefore are likely to be involved in the pathology of narcolepsy.

Behavioral correlates of the activity of serotonergic and non-serotonergic neurons in caudal raphe nuclei.

We investigated the behavioral correlates of the activity of serotonergic and non-serotonergic neurons in the nucleus raphe pallidus (NRP) and nucleus raphe obscurus (NRO) of unanesthetized and unrestrained cats. The animals were implanted with electrodes for recording single unit activity, parietal oscillographic activity, and splenius, digastric and masseter electromyographic activities. They were tested along the waking-sleep cycle, during sensory stimulation and during drinking behavior. The discharge of the serotonergic neurons decreased progressively from quiet waking to slow wave sleep and to fast wave sleep. Ten different patterns of relative discharge across the three states were observed for the non-serotonergic neurons. Several non-serotonergic neurons showed cyclic discharge fluctuations related to respiration during one, two or all three states. While serotonergic neurons were usually unresponsive to the sensory stimuli used, many non-serotonergic neurons responded to these stimuli. Several non-serotonergic neurons showed a phasic relationship with splenius muscle activity during auditory stimulation. One serotonergic neuron showed a tonic relationship with digastric muscle activity during drinking behavior. A few non-serotonergic neurons exhibited a tonic relationship with digastric and/or masseter muscle activity during this behavior. Many non-serotonergic neurons exhibited a phasic relationship with these muscle activities, also during this behavior. These results suggest that the serotonergic neurons in the NRP and NRO constitute a relatively homogeneous population from a functional point of view, while the non-serotonergic neurons form groups with considerable functional specificity. The data support the idea that the NRP and NRO are implicated in the control of somatic motor output.