Laryngeal afferent stimulation enhances Fos immunoreactivity in periaqueductal gray in the cat.

The main functions of the larynx are protection of the airways, respiration, and vocalization. Previous studies have suggested a link between the mechanisms controlling vocalization and afferent feedback from the larynx. We inquired whether stimulation of the laryngeal afferents that run in the internal branch of the superior laryngeal nerve (ISLN) activates neurons of the periaqueductal gray (PAG), a midbrain region implicated in vocalization. We counted the number of neurons expressing Fos, the protein product of the immediate early gene c-fos, in the PAG. The counts were done both in experimental cats after electrical stimulation of the ISLN and nonstimulated controls. We also investigated the possible presence of nitric oxide synthase, an enzyme that synthesizes nitric oxide, in PAG neurons that respond to laryngeal afferent stimulation by double labeling for reduced nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase and Fos. Fos expression was significantly greater (P < or = 0.00714) in the lateral and dorsolateral regions of the PAG in the experimental group than in the controls. The Fos-immunoreactive neurons did not contain NADPH-diaphorase, a marker for nitric oxide synthase. Our study suggests that laryngeal afferent stimulation activates neurons in discrete longitudinal columns of the PAG including the regions that have previously been shown to be involved in vocalization, and that these neurons do not contain nitric oxide synthase.     

Nucleus retroambiguus projections to the periaqueductal gray in the cat

The nucleus retroambiguus (NRA) of the caudal medulla is a relay nucleus by which neurons of the mesencephalic periaqueductal gray (PAG) reach motoneurons of pharynx, larynx, soft palate, intercostal and abdominal muscles, and several muscles of the hindlimbs. These PAG-NRA-motoneuronal projections are thought to play a role in survival behaviors, such as vocalization and mating behavior. In the present combined antero- and retrograde tracing study in the cat, we sought to determine whether the NRA, apart from the neurons projecting to motoneurons, also contains cells projecting back to the PAG. After injections of WGA-HRP in the caudal and intermediate PAG, labeled neurons were observed in the NRA, with a slight contralateral preponderance. In contrast, after injections in the rostral PAG or adjacent deep tectal layers, no or very few labeled neurons were present in the NRA. After injection of [(3)H]leucine in the NRA, anterograde labeling was present in the most caudal ventrolateral and dorsolateral PAG, and slightly more rostrally in the lateral PAG, mainly contralaterally. When the [(3)H]leucine injection site extended medially into the medullary lateral tegmental field, labeling was found in most parts of the PAG as well as in the adjoining deep tectal layers. No labeled fibers were found in the dorsolateral PAG, and only a few were found in the rostral PAG. Because the termination pattern of the NRA fibers in the PAG overlaps with that of the sacral cord projections to the PAG, it is suggested that the NRA-PAG projections play a role in the control of motor functions related to mating behavior.     

Anatomical study of the final common pathway for vocalization in the cat

Vocalization, the nonverbal production of sound, can be elicited in many vertebrates by stimulation in several regions of the limbic system but most easily in the caudal periaqueductal gray (PAG). This study shows that a specific cell group in the lateral part of the caudal PAG and in the tegmentum just lateral to it projects bilaterally to the nucleus retroambiguus (NRA) in the caudal medulla oblongata. Similar but much weaker projections are derived from the dorsal PAG. Neurons in the NRA in turn project via a contralateral pathway through the ventral funiculus of the spinal cord to motoneuronal cell groups, innervating intercostal and abdominal muscles. These projections are stronger on the contralateral side, although at lower thoracic and upper lumbar levels, many fibers recross to terminate in the ipsilateral motoneuronal cell groups. In the brainstem NRA neurons project to the motoneuronal cell groups innervating mouth-opening and perioral muscles as well as to motoneurons innervating the pharynx, soft palate, and tongue, and probably to the larynx. All these muscles are active in vocalization. The present anatomical results, combined with the physiological results of others, indicate that the projections from PAG via NRA to vocalization motoneurons form the final common pathway in vocalization. The role of this pathway in the total framework of emotional behavior is discussed.     

Repetitive vocalizations evoked by electrical stimulation of avian brains. IV. Evoked and spontaneous activity in expiratory and inspiratory nerves and muscles of the chicken (Gallus gallus).

The activity in respiratory nerves and muscles in response to electrical stimulation of vocal substrates in the brain and to CO2 stimulation of the respiratory centers was studied in 28 adult chickens. It was found that the same nerves and muscles were active during both vocalization and respiration. Stimulation of vocal substrates resulted in short latency bursting in the expiratory nerves and muscles. As stimulation intensity increased, progressively longer duration bursts composed of numerous subbursts were produced. By relating muscle activity with sound production , such bursting was shown to underlie evoked vocalizations. Background activity in inspiratory nerves and muscles continued uninterruptedly past stimulus onset only stopping when expiratory activity began. Thereafter inspiratory bursting reciprocated with expiratory bursting and was shown to underlie the intervals between vocalizations. The pattern of activity which was evoked by stimulating vocal substrates was found to strongly interact with the pattern of activity evoked by CO2 stimulation of the respiratory system. Simultaneous records of respiratory and tracheal muscles demonstrated that the same information was sent to both groups of muscles during evoked vocalization. Activity in the respiratory muscles was recorded during spontaneous vocalization of a free-moving bird and was found to resemble that recorded from anesthetized birds. Finally the activity of single units in the obex region of the medulla was recorded during electrical stimulation of vocal substrates and during CO2 stimulation of the respiratory system. Rhythmically active units were found only in the medulla. Unit activity paralleled that found in the nerves and muscles. On the basis on the data accumulated, two models of the chicken vocal system are presented. The first is a model of the sound-producing structures of the chicken. The second is a model of the neural machinery which controls the sound-producing structures. The two models are used as a basis for an explanation of the production of voclizations by the chick of the same species.     

Laryngeal reflex regulation: Peripheral and central neural analyses

Because of the many functions (e.g., glottic closure, gagging, coughing, swallowing, speech) in which the internal laryngeal muscles are active, complex regulatory mechanisms must exist to modify and coordinate these activities. Little information is available, however, on these neural mechanisms, and the present study attempted to provide more information by investigating, in the adult cat, the effects of stimuli to various orofacial and upper respiratory tract sites and nerves on 110 single units recorded in individual laryngeal muscles, in the recurrent laryngeal nerve (RLN), and in the brain stem nucleus ambiguus. A particularly powerful excitatory influence on unitary activity related to laryngeal adduction was produced by laryngeal stimuli; in many instances this influence could not be modified by other sensory inputs (e.g., from vagus or glossopharyngeal nerves). Laryngeal stimulation also produced a profound cessation of respiration and suppression of the inspiratory-related activity of laryngeal abductor units. Excitatory and inhibitory effects as a result of interacting various stimuli were also seen in recordings from motoneurons of the RLN and nucleus ambiguus. The effects described show characteristics similar to those noted in other oral-facial muscles and indicate common integrative and regulatory mechanisms. This study in the adult cat provides a basis of comparison for future studies in the kitten where the effects of laryngeal stimulation are augmented to such an extent that they have been implicated in disorders such as the sudden infant death syndrome (SIDS).     

Multifunctional laryngeal motoneurons: an intracellular study in the cat.

We studied the patterns of membrane potential changes in laryngeal motoneurons (LMs) during vocalization, coughing, swallowing, sneezing, and the aspiration reflex in decerebrate paralyzed cats. LMs, identified by antidromic activation from the recurrent laryngeal nerve, were expiratory (ELMs) or inspiratory (ILMs) cells that depolarized during their respective phases in eupnea. During vocalization, most ELMs depolarized and most ILMs hyperpolarized. Some ILMs depolarized slightly during vocalization. During coughing, ELMs depolarized abruptly at the transition from the inspiratory to the expiratory phase. In one-third of ELMs, this depolarization persisted throughout the abdominal burst. In the remainder ("type A"), it was interrupted by a transient repolarization. ILMs exhibited a membrane potential trajectory opposite to that of type A ELMs during coughing. During swallowing, the membrane potential of ELMs decreased transiently at the onset of the hypoglossal burst and then depolarized strongly during the burst. ILMs hyperpolarized sharply at the onset of the burst and depolarized as hypoglossal activity ceased. During sneezing, ELMs and ILMs exhibited membrane potential changes similar to those of type A ELMs and ILMs during coughing. During the aspiration reflex, ELMs and ILMs exhibited bell-shaped hyperpolarization and depolarization trajectories, respectively. We conclude that central drives to LMs, consisting of complex combinations of excitation and inhibition, vary during vocalization and upper airway defensive reflexes. This study provides data for analysis of the neuronal networks that produce these various behaviors and analysis of network reorganization caused by changes in dynamic connections between the respiratory and nonrespiratory neuronal networks.     

The physiological motor patterns produced by neurons in the nucleus retroambiguus in the rat and their modulation by vagal,peripheral chemosensory,and nociceptive stimulation

The nucleus retroambiguus (NRA) is a neuronal cell group in the medullary ventrolateral tegmentum, rostrocaudally between the obex and the first cervical spinal segment. NRA neurons are premotor interneurons with direct projections to the motoneurons of soft palate, pharynx, and larynx in the nucleus ambiguus in the lateral medulla as well as to the motoneurons in the spinal cord innervating diaphragm, abdominal, and pelvic floor muscles and the lumbosacral motoneurons generating sexual posture. These NRA premotor interneurons receive very strong projections from the periaqueductal gray (PAG) in the context of basic survival mechanisms as fight, flight, freezing, sound production, and sexual behavior. In the present study in rat we investigated the physiological motor patterns generated by NRA neurons, as the result of vagal, peripheral chemosensory, and nociceptive stimulation. The results show that the NRA contains phasic respiratory modulated neurons, as well as nonphasic tonically modulated neurons. Stimulation in the various rostrocaudal levels of the NRA generates site‐specific laryngeal, respiratory, abdominal, and pelvic floor motor activities. Vagal and peripheral chemosensory stimulation induces both excitatory and inhibitory modulation of phasic NRA‐neurons, while peripheral chemosensory and nociceptive stimulation causes excitation and inhibition of nonphasic NRA‐neurons. These results are in agreement with the concept that the NRA represents a multifunctional group of neurons involved in the output of the emotional motor system, such as vomiting, vocalization, mating, and changes in respiration.     

A reinvestigation of the analgesic effects induced by stimulation of the periaqueductal gray matter in the rat. I. The production of behavioral side effects together with analgesia

It has been shown that stimulation-produced-analgesia (SPA) in the cat elicited from the periaqueductal gray matter (PAG) is obtained from sites located in the ventral part, particularly the dorsal raphé nucleus (DRN). These data contrast with the numerous studies performed in the rat in which efficient sites seem widely distributed throughout the PAG. These discrepancies led us to reinvestigate SPA from PAG and adjacent structures in the rat. Central stimulation was delivered through bipolar concentric electrodes (one for each animal). Analgesia was evaluated (before and during central stimulation) by measuring the modification in the vocalization threshold induced by electrical tail shocks or by considering the reaction of the animal to pinch. In contrast with the majority of previous studies, these experiments were performed on the totally freely-moving rat. The most striking result was that, in order to obtain analgesia from all regions of the PAG, it was necessary to apply intensities of central stimulation which also triggered other strong behavioral reactions. With intensities of PAG stimulation which did not induce such side effects, very few effective analgesic sites were found (21/129 sites of which 14/83 were strictly located in the PAG). However, it was possible to define two 'pure analgesic regions', both located in the ventral PAG: one centered on the dorsomedial part of the DRN and the other one situated in the ventrolateral PAG. No modification of nociceptive thresholds was observed when stimulating the dorsal and dorsolateral parts of the PAG as well as structures adjacent to these regions; in some rats, an increase in pain reactivity was even noted. When the intensity of central stimulation (applied to the various parts of the PAG) was increased, some stereotyped 'behavioral responses' occurred depending on the location of the stimulation site: motor effects (gnawing, rotation or tremor) in the ventral PAG and aversive effects (flight, jumping and on occasions, distress vocalizations) in the dorsal, dorsolateral PAG and in the ventral region just surrounding the cerebral aqueduct. Under these conditions, analgesia was obtained from practically the entire PAG, the vocalization threshold being increased dramatically on occasions. It must be emphasized that antinociceptive effects associated with other obvious behavioral manifestations (aversive ones) were also obtained from sites located outside the PAG (colliculi and tectum adjacent to the dorsal and dorsolateral PAG).(ABSTRACT TRUNCATED AT 400 WORDS)     

An electrophysiological characterization of the projection from the central nucleus of the amygdala to the periaqueductal gray of the rat: the role of opioid receptors

The midbrain periaqueductal gray (PAG) and the central nucleus of the amygdala (CNA) are both known to be involved in fear and anxiety, analgesia, vocalization, cardiovascular and respiratory changes, and freezing. Anatomical studies have shown that a connection between these two regions exists but little is known about the physiology or the neurochemical constituents of this pathway. The goals of this study were to characterize the projection from the CNA to the PAG using electrophysiological techniques and to determine whether μ- and/or δ-opioid receptors, which play a large role in a majority of the functions of the PAG, are involved in this pathway. Of the 38 PAG cells tested with single shock stimulation of the CNA, 44% responded; of those, 46% were excited and 54% were inhibited. The latency to onset of response for the inhibitory cells (12.71 ± 6.61 ms) was shorter than that of the excitatory cells (22.33 ± 4.04 ms). Forty-six percent of the 129 PAG cells tested with train electrical stimulation of the CNA responded; 44% were excited and 56% were inhibited. Chemical stimulation of the CNA (10 mMd,l-homocysteic acid) produced similar results; 48% (62/128) of PAG cells responded; 45% of cells were excited and 55% were inhibited. The baseline firing rate of the inhibitory cells was significantly higher compared to the excitatory cells. Chemical stimulation of the CNA produced an increase in blood pressure in 12 animals, a decrease in two animals, and had no effect on the blood pressure of 68 animals. The blood pressure changes ranged between 8.5 and 26.3 mmHg with a mean of 16.2 ± 2.2 mmHg. The effect of naloxone (given either on site in the PAG or systemically) on the response to CNA stimulation was tested in 21 cells. Twenty-five percent of the excitatory cells (2/8) and 77% (10/13) of the inhibitory cells were blocked by naloxone with the majority of the blocked cells located in the ventrolateral PAG. It is concluded that: (1) Approximately 50% of cells in the lateral and ventrolateral columns of the PAG respond to CNA stimulation; (2) the inhibitory response is mediated by a faster conducting or a more direct pathway than the pathway that mediates the excitatory response; (3) neurons that are inhibited by CNA stimulation have a significantly higher baseline firing rate than neurons that are excited, suggesting that they may be tonically active interneurons; and (4) at least one link in the CNA-PAG pathway utilizes μ- or δ-opioid receptors.     

Possible roles for GABAergic inhibition in the vocal control system of the zebra finch

The final common output from the telencephalic vocal control system in songbirds is the projection from nucleus RA, which drives respiratory and syringeal muscles via medullary nuclei. We examined the possible role of GABAergic inhibition in RA of adult male zebra finches by micro-injecting bicuculline, an antagonist of inhibitory GABA(A) receptors, while recording simultaneously with multiple microelectrodes. Following bicuculline injection, the normally high spontaneous activity of RA neurons exhibited a pattern of rhythmic bursting lasting up to 30 min. The bursts were often accompanied by involuntary vocalizations: monosyllabic notes resembling calls. Other experiments used microinjections that were below threshold for involuntary vocalization. When the bird sang to a female during a period after the injection, song structure was degraded: song duration was lengthened, noisiness increased, and novel syllables appeared. The results suggest that GABA normally contributes to regulating excitability in RA. When this regulation is blocked, activity increases sufficiently to engage the respiratory and vocal musculature. Synaptic inputs that affect GABAergic interneurons in RA could thus play a role in initiation and control of vocalization. The abnormal vocalizations produced in the presence of bicuculline suggest that GABAergic inhibition may normally help to shape the pattern of learned vocalizations, as well as to regulate overall RA activity.     

Afferent projections to pharynx and soft palate motoneurons: a light and electron microscopical tracing study in the cat

Pharynx and soft palate are muscles for respiration, vocalization, swallowing, and vomiting. In cat, motoneurons innervating pharynx/soft palate are located in the dorsal group of the nucleus ambiguus (dgNA) in the medulla oblongata. In cat, dgNA is the only part of nucleus ambiguus that can be distinguished as a separate cell group, which makes it possible to study its afferent input. In two cats, WGA-HRP injections in dgNA and surrounding tegmentum resulted in retrogradely labeled cells at several levels of the neuraxis. In 170 cases anterograde tracers were injected in areas in which the cells of origin were identified. Results demonstrate that dgNA afferents originate from the tegmentum dorsolateral to the superior olivary complex, medullary ventromedial tegmentum, caudal raphe nuclei, medullary lateral tegmental field, nucleus retroambiguus (NRA), and adjoining tegmentum, extending into the first cervical segment of the spinal cord. In order to determine whether periaqueductal gray (PAG) and parabrachial nuclei (PB) make synaptic contacts with dgNA, ultrastructural studies combined anterograde tracing from PAG, PB, and NRA with retrograde tracing of pharyngeal and soft palate motoneurons. The results showed that PB, but not PAG, projects to the dgNA and that NRA afferent synapses are three times as numerous as those from PB. The morphology of PB and NRA synapses is consistent with excitatory input. In conclusion, pharyngeal and soft palate motoneurons receive their afferents almost exclusively from the pontine and medullary tegmentum and first cervical spinal segment.     

A reinvestigation of the analgesic effects induced by stimulation of the periaqueductal gray matter in the rat. II. Differential characteristics of the analgesia induced by ventral and dorsal PAG stimulation

This study consists of a detailed analysis of the analgesic effects induced by stimulation of the various parts of the periaqueductal gray matter (PAG) in the freely moving rat. In order to characterize the analgesia, two criteria are considered: (1) the evaluation of the degree of analgesia and behavioral side effects evoked during central stimulation; and (2) the presence of post-effects. Central stimulation (50 Hz sine waves) was delivered via bipolar concentric electrodes and analgesia was quantified by the change in the vocalization threshold induced by electrical stimulation of the tail. Within the ventral PAG, the vocalization threshold increased gradually with the intensity of the central stimulation, the degree of analgesia generally being powerful. There was no relationship between the strength of the analgesic effects and the motor disturbances also produced by stimulation of this region. Antinociceptive effects generally disappeared when the stimulation ceased. Only when the intensity of the stimulation was strong enough to induce very powerful analgesic effects were post-stimulation analgesic effects noticed. Within the dorsal and dorsolateral PAG as well as in the ventral region just surrounding the aqueduct, analgesia appeared suddenly, was generally less pronounced and was always concomitant with strong aversive reactions. In contrast with the analgesia from the ventral PAG, marked post-effects were observed. These latter characteristics were also obtained from stimulation of regions located outside the PAG (colliculi, intercollicular commissure and tectum adjacent to the dorsolateral PAG) although these zones were not extensively studied. By consideration of various data in the literature, it is concluded from this study, which clearly distinguishes stimulation-produced-analgesia (SPA) from ventral PAG versus dorsal PAG, that analgesia induced from this midbrain area involves at least two different neuronal substrates. Whilst the ventral PAG seems to be more preferentially involved in pain modulation, the authenticity of 'analgesia' triggered by stimulation of aversive regions (which are widely spread over the PAG) is questioned and proposals to explain the simultaneous appearance of analgesic effects and aversion are considered.     

Differential effects of noxious and non-noxious input on neurones according to location in ventral periaqueductal grey or dorsal raphe nucleus

Nociceptive and non-nociceptive input to the dorsal raphe nucleus (DR) and to the surroundings periaqueductal grey (PAG) was studied in chloralose-anaesthetized rats. Single units in the midbrain responding to electrical stimulation of a coccygeal nerve were recorded with glass micropipettes. A fluorescence histochemical technique was applied to identify recording sites in the DR and PAG.109 DR-units, 141 PAG-units and 95 units from surrounding structures were tested for responsiveness to electrical nerve stimulation. In 53% of the DR-units, but in only 20% of the PAG- and SN-units, ongoing activity was inhibited by electrical stimulation (I-units) while 42% of the PAG- and SN-units but only 24% of the DR-units were electrically excited (E-units).40 E-units and 24 I-units were tested with repeated noxious radiant heat stimuli applied to the tail or hindpaws. 70% of the E-units were excited by heating, and in 54% of the I-units ongoing activity was inhibited by heating. The majority of the former units were located in the PAG, and most of the latter were proven to be DR-neurones. In 7.5% of the E-units and in 12.5% of the I-units the heat effect was in the opposite direction. The findings are discussed in terms of the now well-established role of the PAG-region in the descending control of pain. The properties of the PAG-E-units suggest that this system is involved in a negative feedback circuit by which pain transmission to the CNS limits itself. DR-I-units may be involved via an additional small loop with the PAG to disinhibit the activation of the PAG pain control system.     

The Effects of Periaqueductally Injected Transmitter Antagonists on Forebrain-elicited Vocalization in the Squirrel Monkey

In 15 squirrel monkeys, vocalization-eliciting electrodes were implanted into the following forebrain structures: anterior cingulate cortex, genu of the internal capsule, amygdala, bed nucleus of the stria terminalis, hypothalamus, midline thalamus, inferior thalamic peduncle and periventricular grey. Then, injections of 29 transmitter antagonists were made into the midbrain periaqueductal grey (PAG) and their effects tested on the elicitability of vocalization from the forebrain. Vocalization could be blocked completely with glutamate antagonists. NMDA receptor antagonists as well as kainate/quisqualate receptor antagonists were effective. Facilitatory effects, i.e. a decrease in threshold of forebrain-elicited vocalization, was obtained with GABA-A receptor, glycine and opioid antagonists. The facilitatory effect of the opioid antagonist naloxone was limited to vocalizations expressing aversive emotional states. GABA-A receptor antagonists not only facilitated forebrain-induced vocalization but also produced vocalization themselves, i.e. without concomitant forebrain stimulation. No effects were obtained with antagonists of muscarinic and nicotinic receptors, with the GABA-B receptor antagonist phaclofen and antagonists of the monoamines dopamine, noradrenaline, adrenaline, serotonin and histamine. It is concluded that the PAG represents a crucial relay station of the vocalization-controlling system. In this station, transmission of vocalization-relevant information depends upon the activation of glutamatergic synapses. Inhibitory control is exerted by GABA, glycine and endogenous opioids. Acetylcholine, dopamine, noradrenaline, adrenaline, serotonin and histamine may play a transient modulatory role; forebrain-induced vocalization, however, does not depend upon the cholinergic or monoaminergic activation of PAG neurons.     

Stimulation of the midbrain periaqueductal gray modulates preinspiratory neurons in the ventrolateral medulla in the rat in vivo

The midbrain periaqueductal gray (PAG) is involved in many basic survival behaviors that affect respiration. We hypothesized that the PAG promotes these behaviors by changing the firing of preinspiratory (pre‐I) neurons in the pre‐Bötzinger complex, a cell group thought to be important in generating respiratory rhythm. We tested this hypothesis by recording single unit activity of pre‐Bötzinger pre‐I neurons during stimulation in different parts of the PAG. Stimulation in the dorsal PAG increased the firing of pre‐I neurons, resulting in tachypnea. Stimulation in the medial part of the lateral PAG converted the pre‐I neurons into inspiratory phase‐spanning cells, resulting in inspiratory apneusis. Stimulation in the lateral part of the lateral PAG generated an early onset of the pre‐I neuronal discharge, which continued throughout the inspiratory phase, while at the same time attenuating diaphragm contraction. Stimulation in the ventral part of the lateral PAG induced tachypnea but inhibited pre‐I cell firing, whereas stimulation in the ventrolateral PAG inhibited not only pre‐I cells but also the diaphragm, leading to apnea. These findings show that PAG stimulation changes the activity of the pre‐Bötzinger pre‐I neurons. These changes are in line with the different behaviors generated by the PAG, such as the dorsal PAG generating avoidance behavior, the lateral PAG generating fight and flight, and the ventrolateral PAG generating freezing and immobility. J. Comp. Neurol. 521: 3083–3098, 2013. © 2013 Wiley Periodicals, Inc.     

The effects of periaqueductal gray and nucleus raphe magnus stimulation on the spontaneous and noxious-evoked activity of lateral reticular nucleus neurons in rabbits

In urethane-anesthetized rabbits the effects of periaqueductal gray (PAG) and nucleus raphe magnus (NRM) stimulation on the spontaneous and the noxious-evoked activity of the lateral reticular nucleus (LRN) neurons were studied. The PAG and the NRM stimulating electrodes were located in the optimal sites for suppressing the jaw-opening reflex (JOR) evoked by the tooth pulp stimulation. It was found that the 12% of neurons tested were affected by one or both stmuli. A total of 80 responsive neurons (52% antidromically activated by the cerebellum) were analyzed. Out of these neurons, 31 showed a convergence to both stimuli, 43 responded only to PAG and 6 only to NRM. Noxious heat stimulation of the contralateral foot was effective in altering the activity of 60% of these neurons. The PAG and NRM stimuli modified the noxious-evoked responses in most of these units. While the excitation was the predominant effect on the spontaneous activity (52 cells), the inhibition was predominant on the noxious-evoked activity (29 cells). These results indicate the presence of connections from PAG and NRM to LRN, probably devoted to the processing of the nociceptive information.     

The identification of brainstem neurones projecting to thoracic respiratory motoneurones in the cat as demonstrated by retrograde transport of HRP

Brainstem neurones which project to the immediate vicinity of the spinal motoneurones which supply the intercostal and abdominal respiratory muscles were identified by means of the retrograde transport of horseradish peroxidase (HRP). A combined electrophysiological and histological technique was used in which recording of phasic inspiratory or expiratory motoneurone activity within upper (T3-T4) or lower (T8-T9) thoracic segments was followed by the ion-tophoretic injection of HRP at these recording sites. HRP labelled cells were concentrated in those brainstem regions known to contain phasic respiratory neurones, namely the ventrolateral nucleus of the solitary tract (vl-NTS) or dorsal respiratory group (DRG), the ambiguus complex or ventral respiratory group (VRG) and the parabrachial pontine (PB) nuclei. In 18 cats, 248 cells were labelled in these three respiratory regions of the brainstem while 668 were much more diffusely distributed in other regions of the medulla and pons. The ipsilateral and contralateral contributions within the respiratory regions were respectively; 23%:77% (DRG), 33%:67% (VRG), 95%:5% (PB). These results are considered in the general context of previous electrophysiological and histological findings, but also with particular reference to a related study of the projections from brainstem neurones to the phrenic nucleus [32].     

Electrophysiological analysis of midbrain periaqueductal gray influence on cardiovascular neurons in the ventrolateral medulla oblongata

Stimulation of sites in the rostral or caudoventral periaqueductal gray (PAG) results in substantial increases in mean blood pressure (MBP) and heart rate (HR). The efferent pathways from these PAG subregions possibly include a relay in the ventrolateral medulla oblongata (VLM), where neurons involved in maintaining vasomotor tone are located. Extracellular recordings were made from 21 cardiovascular neurons in the rostral VLM (RVLM) and from 6 cardiovascular neurons in the caudal VLM (CVLM) of the rat. These neurons showed barosensitivity and cardiac rhythmicity. In addition, the activity of 54 noncardiovascular and nonrespiratory units was recorded. Responses to electrical stimulation of sites in the (rostral or caudal) PAG were studied in 16 of the 21 cardiovascular RVLM neurons, the 6 CVLM neurons, and 46 of the 54 noncardiovascular neurons. Eight of the RVLM neurons were excited by rostral PAG stimulation; the poststimulus time histograms showed a constant latency in five units (32 ± 3 ms). This suggests the presence of relatively direct (although not monosynaptic) excitatory pathways from the rostral PAG to cardiovascular neurons in the RVLM, consisting of slowly conducting fibers (0.2-0.3 m/s). Five RVLM neurons did not respond to rostral PAG stimulation. Three units were tested with caudal PAG stimulation: one was excited, one inhibited, and one was unresponsive. The six cardiovascular CVLM neurons did not respond to PAG stimulation. Of the 46 noncardiovascular neurons, 14 cells were excited, 7 inhibited, and 2 cells antidromically activated. These results confirm earlier findings, extending them to the rostral PAG. They supply further evidence for the influence of the PAG on the cardiovascular function-related neuronal circuitry in the VLM.     

Electrophysiological and functional identification of different neuronal types within the nucleus ambiguus in the cat

Extracellular single neuron recording in nucleus ambiguus demonstrates the existence of different neuronal classes within its electrophysiological limits. Laryngeal motoneurons were identified by their antidromic activation from the recurrent laryngeal and vagus nerves. Interneurons were identified by their antidromic activation from the contralateral cervical spinal cord. Stimulation of the ipsilateral superior laryngeal nerve activated synaptically all laryngeal motoneurons recorded but not interneurons. Most motoneurons and all interneurons showed spontaneous discharges during the inspiratory phase of the respiratory cycle. A few expiratory motoneurons were also recorded. In addition to laryngeal motoneurons and interneurons vagal stimulation also activated a population of efferent neurons located in the nucleus ambiguus with firing patterns not related to respiration. Functional implications of described findings are discussed.     

Divergent synaptic influences affecting discharge patterning of genioglossus motor units

Discharge patterns of single motor units were recorded from 5 inframandibular muscles in acutely prepared adult rhesus monkeys and cats. The alterations in interspike interval patterns were studied in genioglossus motor units during respiration, swallowing, laryngeal, and precentral cortex stimulation in animals anesthetized with urethane. Genioglossus motor units were active in both inspiration and swallowing with 87% of the units demonstrating a significant difference in distributions of their first order interspike interval histograms in swallowing as compared to their respiratory pattern. Interaction between several swallows and the prolonged inhibition of respiration (1–7 sec) altered the discharge pattern of a genioglossus unit from its activity during normal respiration. This significant change in the distribution of the interspike intervals of the motor unit in both first order and autocorrelation histograms occurred during the marked increase in intensity and duration of the gross EMG of the genioglossus muscle during the return of respiration. Stimulation of the precentral cortex of the rhesus monkey indicated a third central synaptic influence on the genioglossus motoneurons with a given motor unit discharging in masticatory movements and swallowing. A first order latency histogram demonstrated a fourth synaptic influence with a correlation between the stimulus pulse to the superior laryngeal nerve innervating the laryngeal region and the latency to when a genioglossus motor unit discharged. The latency analysis suggested a vago-hypoglossal reflex from sensory input of the laryngeal region synaptically affecting genioglossus motoneurons by a polysynaptic pathway.