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Membrane potential changes and discharges from 28 laryngeal motoneurons were recorded intracellularly in the caudal nucleus ambiguus of decerebrate, paralyzed and ventilated cats. Electrical activities were recorded from 17 expiratory laryngeal motoneurons (ELMs) with maximal depolarizing membrane potential in early expiration, and from 11 inspiratory laryngeal motoneurons (ILMs) with maximal depolarizing membrane potential in inspiration. Activities during breathing were compared with those observed during fictive coughing and swallowing evoked by electrical stimulation of the superior laryngeal nerves. These non-respiratory behaviors were evidenced in paralyzed animals by characteristic discharge patterns of the phrenic, abdominal nerves and pharyngeal branch of the vagus nerve. We recorded the activity of 11 ELMs and 5 ILMs during coughing in which ELMs, but not ILMs, exhibited increased membrane depolarization and discharge frequencies. Membrane depolarization and discharge frequencies of all ELMs were also significantly increased during swallowing. In addition, membrane depolarization of most ELMs (15/17) was preceded by a short-lasting hyperpolarization due to chloride-dependent inhibitory mechanisms occurring at the onset of swallowing. Out of 10 ILMs tested during swallowing, 7 exhibited membrane depolarization, preceded in 5 cases by a short-lasting hyperpolarization. Discharge frequencies of ILMs were significantly reduced during swallowing. The same pattern of phasic activities of ILMs and ELMs was observed during coughing and breathing, suggesting the involvement of similar excitatory pathways in both behaviors. These results imply that the duration of activation and the discharge frequency of neurons of the central generator for breathing that drive laryngeal motoneurons are enhanced during coughing. During swallowing, in addition to central excitatory mechanisms, laryngeal motoneurons are subjected to an initial inhibition of unknown origin. This inhibition probably contributes to the temporal organization of the swallowing motor sequence.

The distribution of retrogradely doubly labeled brainstem neurons were analyzed in the cat after injection of two different fluorescent markers into the phrenic and abdominal motor nuclei. Diamidino Yellow (DY) was first injected either ipsilaterally or bilaterally into the ventral horn of lumbar spinal cord, and then Fast Blue (FB) into the right ventral horn of cervical spinal cord. Doubly labeled neurons were mainly found in the caudal ventrolateral medulla (retroambiguus region), in the dorsomedial and dorsolateral regions of the nucleus of the tractus solitarius (NTS) and in the raphe nuclei. In addition, doubly labeled neurons were found in the parabrachial and Kölliker-Fuse nuclei. Our results give anatomical evidence that pontine and medullary neurons are the source of a common pathway to both phrenic and abdominal motor nuclei. These neurons might be involved in strain efforts for expulsion such as vomiting or defecation.

In decerebrate, paralyzed, and ventilated cats, we recorded the activity of 100 spontaneously active phrenic motor axons during the increased phrenic discharges characteristic of fictive vomiting (FV) and coughing (FC). During control respiratory cycles, approximately one-half the neurons were recruited in the first decile of inspiration; recruitment continued throughout inspiration. During FV, the duration of phrenic discharge was halved; 20 of 26 motoneurons studied were recruited in the first decile of the burst. During FC, recruitment times did not change compared with control, although the duration of the phrenic burst doubled. Discharge frequencies increased and recruitment order of phrenic motoneurons was virtually unaffected during FC and FV. Limited recruitment of previously inactive neurons in the filaments from which we recorded was found during FV and FC. During FV, 1 previously inactive motoneuron was recruited in 16 filaments containing 25 spontaneously active motor axons. During FC, 3 new motoneurons were recruited in addition to the 64 already active in 35 filaments. Recruitment during FV and FC was absent even when recording from filaments known, on the basis of antidromic activation, to contain inactive motor axons. During FV, 10 of 26 motoneurons began their discharges with doublets (interspike interval < 10 ms); doublets occurred in only 4 of 67 motoneurons during FC. Already active phrenic motoneurons contributed to the intense phrenic activity associated with both respiratory (coughing) and nonrespiratory (vomiting) behavior by increases in discharge frequency, earlier recruitment, and doublets; the contribution of previously quiescent motoneurons remains uncertain.

In decerebrate, paralyzed and ventilated cats, we monitored the intracellular responses of 30 inspiratory neurons of the dorsal respiratory group (DRG) to stimulation of vagal and expiratory muscle (internal intercostal and abdominal) afferents. We hypothesized that the inhibitory effects of stimulation of expiratory muscle afferents, previously reported, would block the excitatory responses of inspiratory neurons of the DRG to vagal stimulation. Although prolonged stimulus trains to expiratory muscle afferents caused respiratory phase-switching, single shocks or short trains elicited no responses in 17 bulbospinal neurons, excitatory responses in 6, and inhibitory responses in 2. Of the 4 propriobulbar neurons tested, 2 had inhibitory responses and 2 did not respond. In only 2 neurons, both bulbospinal, did conditioning stimuli to expiratory muscle afferents block or reduce the excitatory effects of vagal stimulation. These results suggest that interaction of vagal and expiratory muscle afferents, which might account for the absence of a change in inspiratory duration despite increased vagal afferent feedback at elevated end-expiratory lung volumes, does not occur within the DRG.

We used the expression of the immediate-early gene c-fos, a marker of neuronal activation, to localize brainstem neuronal populations functionally related to fictive cough (FC). In decerebrate, paralyzed, and ventilated cats, the level of Fos-like immunoreactivity (FLI) was examined in five groups of animals: (1) controls, sham-operated unstimulated animals; (2) coughing cats, including both animals in which FC was elicited by unilateral electrical stimulation of the superior laryngeal nerve (SLN) and (3) those in which FC was elicited by bilateral SLN stimulation; (4) stimulated–treated cats, in which bilateral SLN stimulation was applied after selective blockade of FC by codeine; and (5) codeine controls, sham-operated unstimulated cats subjected to administration of codeine. Fifteen brainstem structures were compared for numbers of labeled cells. Because codeine selectively blocks FC, brainstem nuclei activated specifically during FC were identified as regions showing increased FLI after FC and significant reductions in FLI after FC suppression by codeine in stimulated–treated cats. In coughing animals, we observed a selective immunoreactivity in the interstitial and ventrolateral subdivisions of the nucleus of the tractus solitarius, the medial part of the lateral tegmental field, the internal division of the lateral reticular nucleus, the nucleus retroambiguus, the para-ambigual region, the retrofacial nucleus, and the medial parabrachial nucleus. FLI in all these nuclei was significantly reduced in stimulated–treated cats. Our results are consistent with the involvement of neurons overlapping the main brainstem respiratory-related regions as well as the lateral tegmental field and the lateral reticular nucleus in the neural processing of laryngeal-induced FC.

Membrane potential changes and/or discharges from 36 inspiratory neurons were recorded intracellularly in the dorsal respiratory group (DRG; i.e., the ventrolateral subdivision of the nucleus tractus solitarii) in decerebrate, paralyzed, and ventilated cats. Electrical activities were recorded from both somata (n = 10) and axons (n = 26). Activities during quiet breathing were compared with those observed during fictive coughing and swallowing evoked by repetitive electrical stimulation of afferent fibers of the superior laryngeal nerve (SLN). These nonrespiratory behaviors were evident in paralyzed animals as characteristic discharge patterns of the phrenic, abdominal, and hypoglossal nerves. Twenty-six neurons exhibiting antidromic action potentials in response to electrical stimuli applied to the cervical (C3-5) spinal cord were classified as inspiratory bulbospinal neurons (IBSNs). These neurons were considered as premotoneurons. The remaining 10 inspiratory neurons (I-NAA) were not antidromically activated by electrical stimuli applied to either cervical spinal cord or ipsilateral cervical vagus. These neurons are thought to be propriobulbar neurons. We recorded the activity of 31 DRG inspiratory neurons (24 IBSNs and 7 I-NAA) during coughing. All but one (a late-recruited IBSN) discharged a burst of action potentials during the coughing-related phrenic nerve activity. Typically, ramp-like membrane depolarization trajectories and discharge frequencies during coughing were similar to those observed during inspiration. We recorded the activity of 33 DRG inspiratory neurons (23 IBSNs and 10 I-NAA) during swallowing. Most (28/33) neurons were briefly activated, i.e., discharged a burst of action potentials during swallowing, but peak discharge frequency decreased compared with that measured during inspiration. The membrane potentials of nine somata exhibited a brief bell-shaped depolarization during swallowing, the amplitude of which was similar to that observed during inspiration. These results suggest that some inspiratory premotoneurons and propriobulbar neurons of the DRG might be involved in nonrespiratory motor activities, even if clearly antagonistic to breathing (e.g., swallowing). We postulate the existence in the medulla oblongata of adult mammals of neurons exhibiting a "functional flexibility".

Intracellular recordings and labelings with horseradish peroxidase (HRP) of expiratory (E) neurons were performed in decerebrate, paralyzed, and ventilated rats. A total of 37 neurons were recorded, from which 4 cells and 1 axon were labeled. They were located in two regions of the ventrolateral medulla. One was in the rostral portion of the nucleus ambiguus just caudal to the facial nucleus, and the other in the nucleus retroambiguus at the level of the caudal medulla. These expiratory neurons had rhythmical changes in membrane potential similar to those reported in cat, i.e., a depolarization in the intervals between phrenic bursts which evolved in an augmenting (E-aug, n = 15), or bell-shaped or ‘plateau’ (E-all, n = 22) pattern until a rapid hperpolarization at the start of inspiration. Both types were hyperpolarized during inspiration by chloride-dependent, inhibitory postsynaptic potenials (IPSPs) which were demonstrated in 17 neurons (10 E-aug and 7 E-all) from which reversal was obtained. Such IPSPs also existed during post-inspiration (stage I of expiration) in 4 of the 10 augmenting E neurons. They were identified by antidromic stimulation or HRP labeling, or both, as bulbospinal neurons (n = 2), cranial motoneurons (n = 4), or not antidromically activated (NAA) neurons (n = 31). All the identified bulbospinal neurons and the motoneurons bulbospinal neuron with ipsilateral axon giving off intramedullary collaterals, and NAA neurons with rostral medullary projections or with axons crossing the midline. These results indicate that the distribution and the electrophysiological properties of the expiratory neurons in the rat medulla are similar to those described in the cat.

Intracellular recordings and labelings with horseradish peroxidase (HRP) of inspiratory neurons were performed in decerebrate, paralyzed and ventilated rats. A total of 58 neurons were located within the ventrolateral medulla. They were identified as bulbospinal neurons (n = 15), cranial motoneurons (n = 9) and not antidromically activated (NAA) neurons (n = 34) by antidromic stimulation or HRP labeling, or both. These inspiratory neurons had rhythmical changes in membrane potentials similar to those reported in cats, i.e. an abrupt depolarization at the onset of phrenic discharge followed by trajectories of depolarization which evolved into augmenting I, bell-shaped I or decrementing I patterns until a rapid repolarization at the start of expiration. All types were hyperpolarized during expiration by chloride-dependent inhibitory postsynaptic potentials (IPSPs) which were demonstrated in 13 neurons from which the reversal was obtained. Such IPSPs were apparent in two waves throughout expiration, an early one in post-inspiration (stage I of expiration) and a late one in late expiration (stage II of expiration). These properties are also similar to those of feline inspiratory medullary neurons. Four labeled bulbospinal neurons had axonal collaterals which were ipsi- and contralateral to the site of their somata. Two of 6 labeled NAA neurons exhibited profuse axonal arborizations within various medullary nuclei. No medullary axonal collateral was seen from 6 labeled motoneurons. These results indicate that even though in the rat a single concentration of inspiratory neurons within the ventrolateral medulla has been demonstrated, there is no fundamental difference in the organization of the inspiratory neuronal network compared to that of the cat.

The topographical neuronal distribution within the rat nucleus ambiguus has been studied with the simultaneous retrograde labeling technique by means of four different fluorochromes injected within the various muscles and/or nerves of the oro-pharyngeal region. This technique has permitted the identification of several types of neurons along the same coronal plane. Most were motoneurons innervating the various muscles of the upper airway, including pharyngeal constrictor, stylopharyngeal, intrinsic laryngeal and the upper portion of the esophagus. Some neurons may have been preganglionic parasympathetic neurons. No evidence of axonal branching of any of the labeled motoneurons or parasympathetic neurons was found.

The location of neurons within the ventral respiratory group (VRG) of rat was mapped following injections of 3 different fluorochrome tracers into different sites known to receive projections from VRG neurons. Injection sites included muscles innervated by the vagus (X) and glossopharyngeal (IX) nerves, and the sites of expiratory activity in the caudal medulla and of inspiratory activity in the spinal cord at the C4 level. Labeling of vagal motoneurons resulting from fluorochrome injections into muscles innervated by X and IX nerves was always ipsilateral to the site of injection. Both propriobulbar and bulbospinal neurons had primarily ipsilateral projections. No double-labeled cell bodies were observed. The cell bodies of the 3 types of neurons, propriobulbar, bulbospinal and vagal/glossopharyngeal, were unevenly distributed along the rostrocaudal axis of the VRG, suggesting a complex mosaic of neurons which regulate respiratory-related functions such as swallowing and vocalization.

We examined the membrane potentials of 63 respiratory neurons in the ventrolateral medulla of decerebrate rats, whose trajectories had the characteristics of the post-inspiratory neurons, i.e. exhibiting hyperpolarization during inspiration, rapid depolarization at end-inspiration and progressive repolarization with a decrementing pattern during the intervals between phrenic bursts. Synaptic responses of 6 post-inspiratory neurons which were tested by stimulation of cervical vagus or superior laryngeal nerves were excitatory. Eleven of these 63 post-inspiratory neurons were labeled by intracellular injection of horseradish peroxidase (HRP). Ten of these 11 labeled neurons were motoneurons since their axons exited the medulla after joining the roots of cranial nerves. However, only one of these motoneurons was antidromically activated by stimulation of the ipsilateral cervical vagus nerve. We assumed that most of the post-inspiratory medullary neurons of the present study were motoneurons, but not interneurons, although antidromic invasion was not possible after stimulation of the cervical vagus and superior laryngeal nerves. Two post-inspiratory neurons of this sample had bulbospinal axons, which were revealed by antidromical activation of spinal cord and HRP labeling, respectively. The axon of the labeled bulbospinal neuron had axonal collaterals which were distributed within the region of the nucleus ambiguus of the ipsilateral medulla. The functional significance of this type of post-inspiratory neuron is discussed.

We analyzed the membrane potential of 161 respiratory neurons in the medulla of decerebrate rats which were paralyzed and ventilated. Three types of inspiratory (I) neurons were observed: those displaying progressive depolarization in inspiration (augmenting I neurons), those which gradually repolarized after maximal depolarization at the onset of inspiration (decrementing I neurons) and those exhibiting a plateau or bell-shaped membrane potential trajectory throughout inspiration (I-all neurons). Three types of expiratory (E) neurons were also encountered: those in which the membrane potential progressively depolarized (augmenting E neurons), those in which the membrane potential repolarized during the interval between phrenic bursts (decrementing E or post-I neurons) and those exhibiting a plateau or bell-shaped membrane potential trajectory throughout expiration (E-all neurons). Axonal projections of these medullary neurons were identified in the cranial nerves (n = 34), or in the spinal cord (n = 19) as revealed by antidromic stimulation and/or by reconstruction following horseradish peroxidase (HRP) labeling. The other 108 neurons were not antidromically activated (NAA) by the stimulations tested, or had their axons terminating inside the medulla as revealed by HRP labeling. All these respiratory neurons, except for 3 which were hypoglossal motoneurons, had their somata within the ventrolateral medulla, in the region of the nucleus ambiguus, homologous to the ventral respiratory group (VRG) of the cat. No dorsal respiratory group (DRG) was detected within the medulla of the rats. Due to this absence of a DRG, it is concluded that the neural organization of respiratory centers is quite different in cats and rats.

Cancer therapy with cytotoxic drugs such as cisplatin or cyclophosphamide is usually associated with violent crisis of vomiting. Recently, it was shown that 5-HT 3 receptor antagonists block cisplatin-induced vomiting but the mechanisms and their sites of action remain unknown. We tested the hypothesis that these agents act on structures within the central nervous system by evaluating the effectiveness of vagal stimulation in eliciting fictive vomiting in decerebrate, paralyzed and ventilated cats before and after administration of such agents. Fictive vomiting was defined as a series of large bursts of synchronous activity in the phrenic and abdominal (expiratory) nerves (retching) followed by a burst in which the abdominal activity was prolonged (expulsion). The latency and number of these co-activations were measured before and after intravenous administration of three 5-HT 3 receptor antagonists (GR 38032F (Ondansetron), Zacopride, and BRL 43694A (Granisetron)). All compounds, administered at doses of 1 and 2 mg/kg failed to block vomiting behaviour in 100% and 68% of trials, respectively. Nor did their administration affect the latency and number of co-activations. We conclude that intravenous administration of 5-HT 3 receptor antagonists do not act centrally on either the brainstem neuronal network known as the “vomiting center” or related neuronal structures. Our results suggest that the anti-emetic effect of 5-HT 3 receptor antagonists in cisplatin-induced vomiting is mediated peripherally rather than centrally.

Activities of respiratory laryngeal and oropharyngeal respiratory nerves were studied during fictive vomiting elicited by supradiaphragmatic vagus nerve stimulation in the decerebrate cat. Inspiratory laryngeal nerves were strongly inhibited throughout the retching and expulsion phase. Glossopharyngeal, hypoglossal and expiratory laryngeal nerves were coactivated with the phrenic and abdominal nerve bursts. The pharyngeal branch of the vagus nerve discharged during the phrenic and abdominal inter-burst of the retching phase, and was silent during the abdominal expulsion. These activities permit speculation about the role of upper airway muscles during vomiting.

In decerebrate cats, stable intracellular recordings were made from 37 laryngeal motoneurones, the membrane potentials of which varied in relation to respiration. These motoneurones were identified as laryngeal since all were antidromically activated by stimulation of the recurrent laryngeal nerve, but in two, the antidromic activity could only be elicited by vagal stimulation (vagotomized cats). The cell bodies were all located within the nucleus ambiguus. Sixteen cells were depolarized during the phrenic burst and were classified as inspiratory laryngeal motoneurones (ILM). Thet repolarized at end-inspiration and received two successive waves of postsynaptic inhibition during expiration: an early, strong one and a late (end-expiratory), weaker one. The decay of the first wave was related to the duration of postinspiratory phrenic activity. Twenty-one cells depolarized abruptly in early expiration followed by a more-or-less gradual repolarization. They were classified as expiratory laryngeal motoneurones (ELM). All ELM were strongly inhibited during inspiration. Some of them received weak inhibition during end expiratory phase. The rapid and large depolarization observed during early expiration (and consequent maximal discharge frequency) can be explained by two summating mechanisms: a postinhibitory rebound resulting from the removal of inhibition during inspiration, and an excitatory phenomenon of unknown origin. The amplitude of this excitatory phenomenon was largest in cats with the most residual (early expiratory) phrenic activity. To explain the hyperpolarizations occurring in ELM during late expiratory and inspiratory phases and those occurring in ILM during early expiration, we hypothesize that reciprocal inhibition exists between networks controlling ILM and ELM activities or between these motoneurones themselves. The cause of the relationship between residual phrenic nerve activity and the events in early expiration in both ILM (hyperpolarization) and ELM (depolarization) remains to be elucidated. Our observations are consistent with the braking effect on expiratory flow in postinspiration caused by the activation of the adductor muscles driven by the expiratory laryngeal motoneurones, and the release of the abductor muscles driven by the inspiratory laryngeal motoneurones.

We have studied in cats the discharge pattern in response to lung inflation and deflation of 283 medullary respiratory neurones, 173 being inspiratory and 110 expiratory. The ventral respiratory nucleus, near the nucleus ambiguus, was particularly investigated. The neurones were classified into bulbo-spinal neurones, laryngeal motoneurones and propriobulbar neurones by antidromic invasion from the spinal cord or the vagus nerve (collision test). The bulbo-spinal neurones responded in the same direction as spinal motoneurones in the Hering-Breuer reflexes: depression of inspiratory neurones and facilitation of expiratory neurones by inflation of the lungs. All the expiratory laryngeal motoneurones and some inspiratory laryngeal motoneurons responded in the opposite direction to the Hering-Breuer reflexes: depression of expiratory motoneurones and facilitation of inspiratory motoneurones. The function of propriobular neurones could be inferred from their response to Hering-Breuer reflexes: one group of propriobulbar neurones responded in such a manner as to be linked to the activity of the bulbo-spinal neurones; the other group responded in accordance with the activity of the laryngeal motoneurones.

In anesthetized cats, electrical stimulation of the cerebral cortex and subcortical structures with short trains could induce onset or termination of activity of the phrenic nerve. Onset of phrenic discharge was obtained following stimulation of the primary sensory areas of the cerebral cortex, the periaqueductal gray, the ponto-mesencephalic reticular formation, the parabrachial nucleus, the vestibular nucleus, and the fastigial nucleus. When these structures were stimulated by repetitive trains at frequencies of 1–5 Hz, the respiratory rhythm was entrained to these same frequencies. The phrenic discharge during these repetitive stimulations exhibited a decrementing pattern similar to that during thermal polypnea. Termination of phrenic activity was obtained upon stimulation of the motor and visceral cortices, the internal capsule, the pyramidal tract, the red nucleus, the rubrospinal tract and the motor nucleus of the trigeminal nerve. Stimulation of these latter structures by repetitive trains resulted in a temporary or permanent arrest of phrenic discharge. These results are discussed in the context of the possible roles of medullary respiratory neurons in the onset or termination of inspiratory activity.

Intracellular recordings of the activities of 10 inspiratory bulbospinal neurones of the medulla were performed in decerebrate cats. Fictive vomiting was induced by repetitive stimulation of the supra-diaphragmatic vagus nerves and was defined by series of synchronous large bursts in both the phrenic (inspiratory) and abdominal (expiratory) nerves. During these synchronous bursts the inspiratory bulbospinal neurones of both the dorsal and ventral respiratory groups were strongly hyperpolarized by chloride-dependent inhibitory postsynaptic potentials (IPSPs). We concluded that during vomiting the central pattern generator is inhibited, and that another pattern generator drives the spinal respiratory motoneurones.

Intracellular recordings of the activities of 16 bulbar expiratory neurones of the rostral medulla were performed in decerebrate cats. Seven of these identified neurones were intracellularly injected with horseradish peroxidase for morphological examination. We observed 3 categories of expiratory neurones including two pharyngeal motoneurones of the retrofacial nucleus, one with an augmenting, the other with a decrementing discharge pattern. Augmenting patterns were also observed in neurones ventromedial to the retrofacial nucleus and in another located 320 μm from the ventral surface of medulla. Their possible functions are discussed in relation to their anatomical location and morphology.

The central distributions of efferent and afferent components of the pharyngeal branches of the vagus (PH-X) and glossopharyngeal (PH-IX) nerves in the cat were studied by soaking their central cut ends in a horseradish peroxidase (HRP) solution. HRP-labelled PH-X neurones were distributed ipsilaterally in the rostral part of the nucleus ambiguus (NA) and the retrofacial nucleus (RFN); HRP-labelled PH-IX neurones were found in the ipsilateral RFN and the bulbopontine lateral reticular formation (RF). Vagal pharyngeal neurones constituted a large population of brainstem motoneurones. The population of HRP-labelled glossopharyngeal neurones was divided into two components. Indeed, on the basis of their location and somal morphology, the most ventral cells were identified as cranial motoneurones and those scattered in the lateral RF as parasympathetic preganglionic neurones. Application of HRP to the PH-IX nerve resulted also in the labelling of fibres and terminals in the alaminar spinal trigeminal nucleus and the nucleus of the solitary tract (NTS). The afferent fibres entered the lateral medulla with the glossopharyngeal roots, ran dorsomedially, then turned caudally toward the NTS and the caudal part of the alaminar spinal trigeminal motor (V) nucleus. In the NTS, labelled fibres ran mainly along the solitary tract, projecting to terminals in the dorsal and dorsolateral nuclei of the NTS.

In anesthetized or decerebrated cats, extracellular activities of pairs of respiratory neurons located in the regions of the dorsal (DRN), ventral (VRN) and retrofacial (RFN) medullary respiratory nuclei were recorded using two separate microelectrodes. Neurons were classified as bulbospinal or laryngeal if stimulation of the spinal cord or vagus nerve elicited antidromic action potentials, or as propriobulbar if they were not antidromically activated. Of 163 pairs of single unit activities, either inspiratory (143 pairs) or expiratory (20 pairs), cross-correlation analyses indicated that 23% had short latency peaks, either broad (12%) or sharp (1%) in their cross-correlograms, 3% had short latency troughs and 74% had flat cross-correlograms. When the two neurons were located in the DRN (68 pairs) the probability of obtaining a positive cross-correlogram was high for inspiratory bulbospinal neurons, indicating shared inputs and excitatory relationships. When one neuron of the pair was located in the RFN and the other in either the DRN or VRN (95 pairs), cross-correlation analysis revealed shared inputs, excitatory and inhibitory relationships. Among expiratory neurons interactions were only inhibitory with a more frequent incidence (3/20) than between inspiratory neurons (2/143). Our results indicate that: short time scale synchrony due to shared inputs (broad peaks) are largely distributed in the respiratory neuronal network and operate over long distance (i.e. RFN, caudal medulla); excitatory coupling may exist between remote neurons but is more frequent between inspiratory bulbospinal neurons located in the DRN.

1. We recorded the membrane potentials of sixty-three respiratory neurones in the rostral, ventral medulla of decerebrate vagotomized cats. Stable recordings were obtained in thirty-eight expiratory and twenty-five inspiratory neurones. Axonal projections were identified by antidromic invasion after electrical stimulation of the region of the dorsal respiratory group (DRG), spinal cord, and the cervical vagus, superior laryngeal and pharyngeal nerves. 2. Two types of expiratory neurones were encountered: those in which the membrane potential progressively depolarized (augmenting neurons, n = 22) and those in which the membrane potential repolarized (decrementing or post-inspiratory neurones, n = 16) during the interval between phrenic bursts. Both types were hyperpolarized during inspiration by chloride-dependent, inhibitory postsynaptic potentials (IPSPs) which decreased membrane resistance. In augmenting neurones two waves of IPSPs appeared, one early and one late in inspiration. 3. Five out of seventeen augmenting expiratory neurones tested were antidromically activated by contralateral stimulation of the spinal cord (n = 3) or the DRG (n = 2). Spinal axons were not detected in any of the sixteen decrementing expiratory neurones tested. Of thirteen expiratory neurones tested with pharyngeal nerve stimulation, one (an augmenting type) was antidromically activated. Superior laryngeal or vagal axons could not be demonstrated for any expiratory neurones. 4. Two types of inspiratory neurones were also encountered: those displaying progressive depolarization throughout inspiration (n = 5) and those which gradually repolarized after maximal depolarization at the onset of inspiration (n = 10). None of the former had identifiable spinal or medullary axons, but superior laryngeal axons were demonstrated in three and pharyngeal axons were found in three. None of the latter was antidromically activated from any of the sites stimulated. 5. Stimulation of the superior laryngeal or pharyngeal nerves evoked excitatory postsynaptic potentials (EPSPs) in all neurones except in post-inspiratory neurones. In these, stimulation of the superior laryngeal or pharyngeal nerves evoked IPSPs in five of twelve neurones tested. 6. We conclude that a spectrum of respiratory neurones lie within or ventral to the retrofacial nucleus. These neurones may control upper-airway muscles or may play a role in chemoreception.

In decerebrate, paralyzed and artificially ventilated cats, we recorded the discharge of 64 motor axons supplying the pharyngeal muscles. Filaments containing motor axons, with discharges related to the respiratory cycle (phrenic nerve activity), were teased from the pharyngeal branches of the vagus and glossopharyngeal nerves. Most units (n = 41) fired only during expiration and exhibited a steady, a decreasing or a late augmenting discharge pattern. These units were found only in vagal filaments. Twenty three units discharged during inspiration and exhibited a steady, a late augmenting or a tonic discharge pattern. The inspiratory-related units were present in both the vagus (n = 13) and glossopharyngeal (n = 10) nerves. Nineteen of 20 pharyngeal inspiratory-related units tested were activated at short latency (range 3.4 to 8.0 ms) by stimulation of afferents in the superior laryngeal nerve (SLN). In 13 of these, such stimulation also suppressed their spontaneous activity, SLN stimulation elicited in all 17 pharyngeal expiratory-related units tested a short latency (range 0 to 8 ms) reduction of activity, followed in 7 units by an increase in activity. SLN stimulation occasionally evoked single or rhythmic multifibre bursts in the vagal pharyngeal filaments. These bursts, involving expiratory-related units, likely correspond to the buccopharyngeal stage of swallowing.

The effects of inhibitory (γ-aminobutyric acid (GABA) and glycine) and excitatory ( l -glutamate and dl -homocysteate, DLH) amino acids on the excitability of respiratory bulbospinal neurons were studied in decerebrate, paralyzed, bilaterally vagotomized, artificially ventilated cats. Unit activities were recorded extracellularly in the medulla in both the ventrolateral portion of the nucleus tractus solitarius and the para-ambigual region in the vicinity of the nucleus ambiguus (dorsal and ventral respiratory groups, respectively). All neurons were bulbospinal since they could be antidromically activated by electrical stimuli to the spinal cord. We used variations in antidromic latency (ADL) as a measure of changes in excitability of the soma. All neurons exhibited variations in ADL related to the respiratory cycle, being shortest (minimum ADL) during neural activity and longest (maximum ADL) in the silent period. Neurons whose discharge frequencies fell during application of putative inhibitory amino acids showed an increase of minimum ADL compared to control, indicating hyperpolarization. Minimum ADL, in some cells, became shorter during application of excitatory amino acids, indicating depolarization; in others, mechanisms secondary to increased neuronal firing likely obscured their effects. The transient maximum ADL usually present at the onset of the silent period was increased by excitatory amino acids and, in some units, was reduced or eliminated by inhibitory amino acids. These effects are discussed in terms of a modulation by synaptic inputs and neurotransmitters of the cumulative afterhyperpolarization which follows bursts of action potentials.

Activities of respiratory laryngeal motoneurones were recorded intracellularly in the nucleus ambiguus of the cat. Some of them were intracellularly injected with peroxidase for morphological reconstruction. Stimulation of the superior laryngeal nerve (SLN) evoked excitatory responses in both somata and axons of expiratory laryngeal motoneurones. In inspiratory laryngeal motoneurones, the responses induced by the SLN depended on the site of recording: inhibition and decrease of excitatory input in somata, excitation in axons. We conclude that excitatory synaptic effects of SLN stimulation acts mainly on, or close to, the initial segment of inspiratory motoneurones, while inhibition reaches the somato-dendritic region.

The influence of halothane anesthesia on respiratory neurons of the region of the retrofacial nucleus were studied in decerebrate, bivagotomized, paralyzed and artificially ventilated cats. Compared with the control situation without anesthesia, the infusion of halothane induced an important decline of spike activity of inspiratory neurons, at times abolishing their activity, but had no significant effect on the expiratory neurons. The functional significance of these findings is briefly discussed.