Inspiratory on-switch evoked by stimulation of the mesencephalon: Activity of phrenic and laryngeal motoneurones

Summary In anaesthetized cats (chloralose-urethan) the effects of brief tetanic electrical stimulation (50 to 100 ms) of the mesencephalic central gray matter and reticular formation on the inspiratory on-switch were studied during the expiratory (E) phase on the gross and unitary activities of phrenic, laryngeal inspiratory and laryngeal expiratory nerves. On the inspiratory laryngeal and phrenic nerves, stimulation elicited a short latency gross response concomitant with the train: the inspiratory Primary Response (Prim.R.) which is followed by an inspiratory Patterned Response (Patt.R.) of longer duration which corresponded to the inspiratory on-switch. The Patt.R. generally appeared from the Prim.R. within a latent period (Silent Phase: Sil.P.) as long as 100 ms. On the expiratory laryngeal nerve, stimulation elicited a brief activation (expiratory Prim.R.) concomitant with the beginning of the inspiratory laryngeal Prim.R. and which rapidly stopped as the latter continued during the stimulus train. The inspiratory Prim.R. corresponded to a simultaneous activation of both early and late (so defined during their spontaneous discharge) inspiratory motoneurones. The laryngeal motoneurones were more strongly activated than the phrenic ones. During the inspiratory Patt.R. all the phrenic motoneurones presented a recruitment delay earlier, compared with the spontaneous one, whereas the recruitment drastically changed from an inspiratory laryngeal motoneurone to another. Thus, the two pools of motoneurones presented different properties of activation. During the inspiratory Sil.P. no concomitant expiratory laryngeal activation was observed when most of the inspiratory motoneurones were inactive. As some inspiratory laryngeal motoneurones did not stop firing, the existence of some central respiratory neurones exhibiting a similar persistent activity and subserving the inspiratory on-switch mechanisms may be hypothesized.Supported by CNRS (LA 205 and ATP n 4188) and Fondation pour la Recherche Médicale     

Inspiratory on-switch evoked by stimulation of mesencephalic structures: A patterned response

The effects of brief tetanic electrical stimulation (50 to 100 ms) of the mesencephalic central gray matter and reticular formation on the inspiratory "on-switch" mechanism were studied during expiration (E phase) in cats anaesthetized with urethan-chloralose. Stimulation during the E phase evoked powerful effects on the phrenic nerve discharge comprising (1) a primary response (Prim. R.) during the train; (2) a patterned response (Patt. R.) resembling that of the normal inspiratory (I) phase and lasting 170 to 1,000 ms. The patterned response corresponded to activation of the inspiratory on-switch (E-I switching) and appeared either immediately after the Prim. R. or within a latent period of 100 ms. The primary response was always obtained during the E phase whatever the stimulation intensity (0.1-1.0 mA). The patterned response was a function (a) of the stimulus time: the later the stimulus in the E phase, the longer the duration of the response; (b) of the stimulus intensity: with 1.0 mA current strength the response was obtained throughout the E phase; with weaker stimuli (0.4 to 0.5 mA) the response was always evoked by trains delivered early (0-300 ms) or late (1,000-1,800 ms) in the E phase; while it occurred irregularly to stimulation in the middle (300-1,000 ms) of the E phase. These results indicate that: (a) the system which promotes inspiration is progressively rather than abruptly depressed at the beginning of expiration, followed by a progressive inspiratory-promoting facilitation at the end of expiration; (b) the patterned response, mostly independent of the stimulus parameter, is not stereotyped and displays considerable plasticity.     

Chemical activation of caudal medullary expiratory neurones alters the pattern of breathing in the cat.

1. The purpose of this work was to ascertain whether the activation of caudal expiratory neurones located in the caudal part of the ventral respiratory group (VRG) may affect the pattern of breathing via medullary axon collaterals. 2. We used microinjections of DL-homocysteic acid (DLH) to activate this population of neurones in pentobarbitone-anaesthetized, vagotomized, paralysed and artificially ventilated cats. Both phrenic and abdominal nerve activities were monitored; extracellular recordings from medullary and upper cervical cord respiratory neurones were performed. 3. DLH (160 mM) microinjected (10-30 nl for a total of 1.6-4.8 nmol) into the caudal VRG, into sites where expiratory activity was encountered, provoked an intense and sustained activation of the expiratory motor output associated with a corresponding period of silence in phrenic nerve activity. During the progressive decline of the activation of abdominal motoneurones, rhythmic inspiratory activity resumed, displaying a decrease in frequency and a marked reduction or the complete suppression of postinspiratory activity as its most consistent features. 4. Medullary and upper cervical cord inspiratory neurones exhibited inhibitory responses consistent with those observed in phrenic nerve activity, while expiratory neurones in the caudal VRG on the side contralateral to the injection showed excitation patterns similar to those of abdominal motoneurones. On the other hand, in correspondence to expiratory motor output activation, expiratory neurones of the Bötzinger complex displayed tonic discharges whose intensity was markedly lower than the peak level of control breaths. 5. Bilateral lignocaine blockades of neural transmission at C2-C3 affecting the expiratory and, to a varying extent, the inspiratory bulbospinal pathways as well as spinal cord transections at C2-C3 or C1-C2, did not suppress the inhibitory effect on inspiratory neurones of either the ipsi- or contralateral VRG in response to DLH microinjections into the caudal VRG. 6. The results show that neurones within the column of caudal VRG expiratory neurones promote inhibitory effects on phrenic nerve activity and resetting of the respiratory rhythm. We suggest that these effects are mediated by medullary bulbospinal expiratory neurones, which may, therefore, have a function in the control of breathing through medullary axon collaterals.     

Switching of the respiratory phases and evoked phrenic responses produced by rostral pontine electrical stimulation

1. In midcollicular-decerebrate, gallamine-paralysed, vagotomized cats, efferent phrenic discharge was recorded as an indicator of the central respiratory cycle. Electrical stimulation (50-250/sec) delivered in the rostral lateral pontine ;pneumotaxic centre' region (in and near nucleus parabrachialis), and set to occur at specified times in the cycle, produced powerful respiratory effects: (a) at dorsolateral points, inspiratory-facilitatory effects (increase of phrenic discharge, shortening of the expiratory phase); (b) at ventrolateral points, expiratory-facilitatory effects (decrease of phrenic discharge, shortening of the inspiratory phase, lengthening of the expiratory phase).2. At both inspiratory-facilitatory and expiratory-facilitatory points, a single stimulus delivered during the inspiratory phase produced a short-latency (4-7 msec) reduction of phrenic discharge, followed by a wave of increased activity. The short latency of the response indicates the existence of paucisynaptic descending inhibitory pathways. Succeeding stimuli in a high-frequency train produced alternating waves of evoked activity and depression; the form of the responses depended on stimulus frequency and on locus of stimulation.3. At inspiratory-facilitatory points, short stimulus trains (10-30 stimuli) of adequate strength delivered in the middle and late expiratory phase caused early termination of the phase (latency 100-300 msec) and switching to a complete inspiratory phase, in which the phrenic discharge pattern resembled that in a normal inspiratory phase. Similarly, adequate stimulus trains applied at expiratory-facilitatory points during the middle and late inspiratory phase caused early termination of the phase and switching to a complete expiratory phase.4. The threshold for occurrence of each type of phase-switching response depended on stimulus current, frequency, number of stimuli, and time of stimulus delivery. As stimulus trains were delivered later in the phase, the threshold for switching to the succeeding phase was progressively reduced. Moreover, the nature of the evoked effects was a non-linear function of stimulus characteristics: a small increase of stimulus efficacy changed the system's response from (a) moderate shortening of the phase or transient change in phrenic discharge, to (b) complete termination of the phase.5. These results indicate that, as each respiratory phase progresses, there is a steady increase of excitability in systems which promote the onset of the succeeding phase. Further, the existence of a relatively sharp threshold for switching of the respiratory phases suggests that the phase transitions occur when critical levels of excitation and inhibition are reached synchronously in populations of respiratory neurones.     

Converse motor output of inspiratory bulbospinal premotoneurones during vomiting

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.     

Activity of Brainstem Respiratory Neurones just before the Expiration-Inspiration Transition in the Rat

Inspiratory activity of the hypoglossal nerve (XIIn) often precedes that of the phrenic nerve (PHRn). By manipulating artificial respiration, this preceding activity (pre-I XIIn activity) can be lengthened or isolated prematurely (decoupled XIIn activity) without developing into overt PHRn-associated inspiratory bursts. We hypothesized that these pre-I and decoupled XIIn activities, collectively termed 'XIIn-w/o-PHRn activity', reflect certain internal states of the respiratory centre at the period just prior to the transition from the expiratory phase to the inspiratory phase. In decerebrate, neuromuscularly blocked and artificially ventilated rats, the firing properties of medullary respiratory neurones were examined during the period of the XIIn-w/o-PHRn activity. The majority of the inspiratory neurones examined could be classified into two types: one was active (XIIn-type) and the other was inactive (PHRn-type) during the XIIn-w/o-PHRn period. On the other hand, augmenting expiratory (E-AUG) neurones of the Bötzinger complex (BOT) and the caudal ventral respiratory group (VRG) fired intensively during this period. Their firing stopped at the onset of the overt inspiratory bursts in the XIIn and PHRn, suggesting that BOT E-AUG neurones inhibit PHRn-type, but not XIIn-type, inspiratory neurones. We hypothesize that XIIn-type inspiratory activity facilitates the phase change from expiration to inspiration, through activation of certain inspiratory neurones that inhibit the firing of BOT E-AUG neurones and generation of the overt inspiratory bursts in XIIn-type and PHRn-type inspiratory neurones.     

Bötzinger-complex, bulbospinal expiratory neurones monosynaptically inhibit ventral-group respiratory neurones in the decerebrate rat

Extracellularly recorded action potentials from 49 Bötzinger-complex, bulbospinal expiratory neurones were used as triggers to compute 162 spike-triggered averages (STAs) of intracellular potentials recorded from 167 respiratory neurones in the ventral respiratory group (VRG) near the obex in 15 vagotomized, paralysed, ventilated and decerebrated rats. All of the Bötzinger-complex expiratory neurones were antidromically activated from the ipsilateral border between the C2/C3 segments of the spinal cord and discharged only during the late part of expiration with an augmenting pattern. We found evidence for monosynaptic inhibitory post-synaptic potentials (IPSPs) in 74 (～44%) of the STAs computed using 34 (～69%) of the trigger neurones. For vagal motoneurones, IPSPs were found in 24 of the 53 STAs of expiratory motoneurones, but in none of the 12 STAs of inspiratory motoneurones. For inspiratory neurones, IPSPs were found in 23 of the 33 STAs of bulbospinal neurones and in 6 of the 26 STAs of not antidromically activated (NAA) neurones. For expiratory neurones, IPSPs were found in one of the two STAs of bulbospinal neurones and in 20 of the 36 STAs of NAA neurones. We conclude that Bötzinger-complex, bulbospinal expiratory neurones monosynaptically inhibit bulbospinal inspiratory neurones, expiratory vagal motoneurones and other unidentified inspiratory and expiratory neurones in the VRG of rats during the late part of expiration.     

Influence of rubrospinal tract and the adjacent mesencephalic reticular formation on the activity of medullary respiratory neurons and the phrenic nerve discharge in the rabbit

Suprapontine brain sites acting on the central respiratory system have been demonstrated to give rise to inspiratory as well as expiratory facilitatory effects. In the present study the inspiratory inhibitory effect which has been reported in the cat to be elicited consistently by electrical stimulation of the rubrospinal tract and the adjacent mesencephalic reticular formation was examined in the urethane-anaesthetized rabbit. Stimulation of these sites with single electrical shocks of moderate intensity induced a short latency (onset after 3.0 ms) transient (duration: 29 ms) inhibition of the phrenic nerve activity (PHR). Short volleys of stimuli applied in mid- to late-inspiration led to a premature off-switch of inspiration. The extracellularly recorded discharge activity of the different types of medullary respiration-related units (RRU) reflected these alterations, accordingly. Axonal connections of RRU with mesencephalic structures were evaluated. Examination of orthodromic responses of medullary RRU to stimulation of this pathway revealed that most bulbospinal inspiratory neurons (10 out of 13) were paucisynaptically inhibited after short latency (at least 1.2 ms). The conduction time from bulbospinal inspiratory neurons to the recording site of PHR was 1.6 ms. Thus, a disynaptic pathway — including bulbospinal inspiratory neurons — is suggested inducing inspiratory inhibition 3.0 ms after single shock midbrain stimulation. This inhibition results in disfacilitation of phrenic motoneurons. The fact that extensive electrolytic lesions of the pneumotaxic center in rostral pons did not abolish the observed inspiratory inhibitions excludes these structures from being involved. A direct pathway from the red nucleus and the adjacent reticular formation to phrenic nuclei of the spinal cord, however, can not be excluded from being involved in the demonstrated inspiratory inhibition. The described effects may play a role in behavioral or voluntary control of respiration.     

Electrical stimulation of arterial and central chemosensory afferents at different times in the respiratory cycle of the cat: II. Responses of respiratory muscles and their motor nerves

The response patterns of the electrical activity of the respiratory motor nerves and muscles to brief electrical stimulation of the arterial and the intracranial chemosensory afferents were studied in anesthetized cats. Stimulation during inspiration increased the activity of phrenic nerve and the inspiratory muscles (intercostal, diaphragm) with a latency of 15–25 ms, whereas expiratory muscle activity in the following expiration remained almost unaltered. Stimulation during expiration increased the activity of expiratory nerves and muscles (intercostal, abdominal) after a delay of 80–120 ms. The later the stimulation occurred in the insor expiratory period the larger the increase in amplitude and in steepness of rise of the respective integrated activity in respiratory nerves and muscles. Stimulation in early inspiration shortened the discharge period of inspiratory muscles, whereas excitation in early expiration caused an earlier onset and prolonged the activity in the expiratory muscles. Stimulation in the late phase of ins- or expiration prolonged the discharge of the respective nerves and muscles. Both the arterial (carotid sinus nerve, CSN, and aortic nerve, AN) and intracranial chemosensory (VM) afferents stimuli were able to affect both the inspiratory and the expiratory mechanisms. The restriction of the effects to the phase of the stimulus suggests a mechanism by which these afferents, when activated during inspiration, effectively project only to inspiratory neurones, and vice versa for expiration.Supported by the Deutsche Forschungsgemeinschaft, SFB 114 Bionach     

Reorganisation of respiratory network activity after loss of glycinergic inhibition

gamma-Aminobutyric acid (GABA)-ergic and glycinergic inhibition is believed to play a major role in the respiratory network. In the present study we tested whether specific blockade of glycinergic inhibition resulted in changes in respiratory network interaction and function. Using the working heart-brainstem preparation from adult mice, we recorded phrenic nerve activity and the activity of different types of respiratory neurones located in the ventrolateral medulla. Strychnine (0.03-0.3 microM) was given systemically to block glycine receptors (Gly-R). During exposure to strychnine, post-inspiratory (PI) neurones shifted their onset of discharge into the inspiratory phase. As a consequence, the post-inspiratory phase failed and the rhythm changed from a three-phase cycle (inspiration, post-inspiration, expiration, with a frequency of about. 0.24 Hz) to a faster, two-phased cycle (inspiration expiration, frequency about 0.41 Hz). Inspiratory and expiratory neurones altered their augmenting membrane potential pattern to a rapidly peaking pattern. Smaller voltage oscillations at approximately 10 Hz and consisting of excitatory and inhibitory postsynaptic potential sequences occurred during the expiratory interval. Due to their high frequency and low amplitude, such oscillations would be inadequate for lung ventilation. We conclude that, under physiological conditions, glycinergic inhibition does indeed play a major role in the generation of a normal respiratory rhythm in adult mice. After failure of glycinergic inhibition a faster respiratory rhythm seems to operate through reciprocal GABAergic inhibition between inspiratory and expiratory neurones, while phase switching is organised by activation of intrinsic membrane properties.     

Dopamine1 receptor agonists reverse opioid respiratory network depression, increase CO2 reactivity

In adult pentobarbital-anesthetized and unanesthetized decerebrate cats, the D(1)R agonists (6-chloro-APB, SKF-38393, dihydrexidine) given intravenously restored phrenic nerve and vagus nerve respiratory discharges and firing of bulbar post-inspiratory neurons after the discharges were abolished by the micro-opioid receptor agonist fentanyl given intravenously. Reversal of opioid-mediated discharge depression was prevented by the D(1)R antagonist SCH23390. Iontophoresis of the micro-opioid receptor agonist DAMGO depressed firing of medullary bulbospinal inspiratory neurons. Co-iontophoresis of SKF-38393 did not restore firing and had no effect on bulbospinal inspiratory neuron discharges when applied alone. The D(1)R agonists given intravenously prolonged and intensified phrenic nerve and bulbospinal inspiratory neuron discharges. They also increased reactivity to CO(2) by lowering the phrenic nerve apnea threshold and shifting the phrenic nerve-CO(2) response curve to lower et(CO(2)) levels. Intravenous fentanyl on the other hand decreased CO(2) reactivity by shifting the phrenic nerve apnea threshold and the response curve to higher et(CO(2)) levels. Fentanyl effects on reactivity were partially reversed by D(1)R agonists.     

The differential organization of medullary post-inspiratory activities

Membrane potential trajectories of 68 bulbar respiratory neurones from the peri-solitary and peri-ambigual areas of the brain-stem were recorded in anaesthetized cats to explore the synaptic influences of post-inspiratory neurones upon the medullary inspiratory network.A declining wave of inhibitory postsynaptic potentials resembling the discharge of postpinspiratory neurones was seen in both bulbospinal and non-bulbospinal inspiratory neurones, including alpha- and beta-inspiratory, early-inspiratory, late-inspiratory and ramp-inspiratory neurones.Activation of laryngeal and high-threshold pulmonary receptor afferents excited bulbar post-inspiratory neurones, whilst in the case of inspiratory neurones such stimulation produced enhanced postsynaptic inhibition during the same period of the cycle. Activation of post-inspiratory neurones and enhanced post-inspiratory inhibition of inspiratory bulbospinal neurones was accompanied by supression of the after-discharge of phrenic motoneurones.These results suggest that a population of post-inspiratory neurones exerts a widespread inhibitory function at the lower brain-stem level. Implications of such an inhibitory function for the organization of the respiratory network are discussed in relation to the generation of the respiratory rhythm.     

Spinal connections of ventral-group bulbospinal inspiratory neurons studied with cross-correlation in the decerebrate rat

We examined the synaptic connections from ventral-group bulbospinal inspiratory neurons to upper cervical inspiratory neurons and phrenic and intercostal motoneurons in decerebrate rats using cross-correlation. Inspiratory neurons were recorded in the medulla (n=28) at the level of the obex and from the upper-cervical segments (C1 and C2) of the spinal cord (n=29) in 18 vagotomized, paralyzed, ventilated, and decerebrated rats. The neurons were identified by their inspiratory firing pattern and antidromic activation from the spinal cord at C7. Whole-nerve recordings were made using bipolar electrodes from the central cut ends of the C5 phrenic nerve and the external and internal intercostal nerves at various thoracic levels. Cross-correlation histograms were computed between these recordings to detect short time scale synchronizations indicative of synaptic connections. Cross-correlation histograms (n=20), computed between the activities of ventral-group bulbospinal inspiratory neurons and the phrenic nerve, all showed peaks (mean half-amplitude width±SD, 1.1±0.3 ms) at short latencies (mean latency±SD, 2.0±0.6 ms) suggestive of monosynaptic excitation. Cross-correlation histograms (n=33), computed between the activities of ventral-group bulbospinal inspiratory neurons and upper-cervical inspiratory neurons, displayed four (12%) peaks (mean halfamplitude width±SD, 0.9±0.1 ms) at short latencies(mean latency±SD, 1.8±0.6 ms) suggestive of monosynaptic excitation, and six (18%) peaks (mean half-amplitude width±SD, 1.4±0.4 ms) at latencies near zero suggestive of excitation fro m a common source. Cross-correlation histograms (n=34), computed between the activities of ventral-group bulbospinal inspiratory neurons and the internal and external intercostal nerves at various thoracic levels (T2-8), showed six (18%) peaks (mean half-amplitude width±SD, 2.5±0.5 ms) at short latency (mean latency±SD, 4.5±1.1 ms) suggestive of oligosynaptic connections. Cross-correlation histograms (n=42) computed between activities of intercostal nerves at various levels of the thoracic spinal cord showed central peaks suggestive of excitation from a common source. Although the size of the peaks decreased with segmental separation, the displacement of the peaks from time zero did not increase with segmental separation (mean displacement±SD, 0.6±0.6 ms) as would be expected if the common excitation resulted from a descending monosynaptic excitation by a source such as the ventral-groupbulbospinal inspiratory neurons. We conclude that all ventral-group bulbospinal inspiratory neurons make monosynaptic connections to phrenic motoneurons, a few make monosynaptic connections to upper-cervical inspiratory neurons, but connections to intercostal motoneurons are made via interneurons.     

Decrementing expiratory neurons of the Bötzinger complex

Summary In Nembutal-anesthetized, immobilized and artificially ventilated cats, decrementing expiratory (E-DEC) neurons which were excited by lung inflation were isolated in the vicinity of the Bötzinger complex. Then intracellular recordings were made from the respiratory neurons in the contralateral ventral respiratory group (VRG). The intracellular membrane potentials were averaged using extracellular spikes of the E-DEC neurons as triggers (spike-triggered averaging method). Hyperpolarizing potentials locked to the triggering spikes were obtained and they were shown to be unitary IPSPs since their polarity was reversed when averaged during passage of hyperpolarizing current. The latencies of antidromic activation of the E-DEC-neurons from the area of intracellular recordings were shorter by about 0.2 ms than those of unitary IPSPs. This showed that the connections were monosynaptic. A total of 47 pairs were analyzed and unitary IPSPs were found in 12 pairs. The E-DEC neurons inhibited both inspiratory and expiratory neurons, including bulbospinal inspiratory neurons, propriobulbar inspiratory neurons, and vagal motoneurons with expiratory activity. These inhibitory E-DEC neurons, receiving excitatory inputs from the stretch receptors of the lungs, presumably intervene in reflex loops such as the Hering-Breuer reflex and may make some contribution to normal breathing.Supported by grants-in-aid for science research nos. 60304044, 62570068 from the Japan Ministry of Education, Science and Culture     

Effects of hypocapnia on respiratory timing and inspiratory activities of the superior laryngeal, hypoglossal, and phrenic nerves in the vagotomized rat

Effects of acute hypocapnia on respiratory timing (inspiratory and expiratory times (TI, TE) ) and on inspiratory activities of the efferent superior laryngeal (Xs1), hypoglossal (XII), and phrenic (Phr) nerves were studied in artificially ventilated vagotomized, and anesthetized rats. Hyperventilation induced a decrease in respiratory frequency exclusively due to prolongation of TE and resulted in expiratory apnea. Inspiratory activities of three nerves decreased with reduction in CO2 concentration of end-tidal gas (FETCO2), and disappeared simultaneously at a threshold FETCO2 for apnea. The decrease in the peak inspiratory activity by hypocapnia was larger in the XII than in the Phr or Xs1 nerve (XII greater than Phr greater than Xs1). The results suggest that the CO2 stimulus (mainly via a central chemosensor) plays an important role in the process of terminating expiration or of expiratory-inspiratory phase switching and that the responses of the XII or Xs1 motoneurons to variation in CO2 stimulus differ from that of the Phr motoneurons (or of the Phr driving medullary neurons). A possible functional significance of these observations is discussed.     

Synaptic interactions between respiratory neurons during inspiratory on-switching evoked by vagal stimulation in decerebrate cats

To elucidate neuronal mechanisms underlying phase-switching from expiration to inspiration, or inspiratory on-switching (IonS), postsynaptic potentials (PSPs) of bulbar respiratory neurons together with phrenic nerve discharges were recorded during IonS evoked by vagal stimulation in decerebrate and vagotomized cats. A single shock stimulation of the vagus nerve applied at late-expiration developed an inspiratory discharge in the phrenic neurogram after a latency of 79+/-11 ms (n = 11). Preceding this evoked inspiratory discharge, a triphasic response was induced, consisting of an early silence (phase 1 silence), a transient burst discharge (phase 2 discharge) and a late pause (phase 3 pause). During phase 1 silence, IPSPs occurred in augmenting inspiratory (aug-I) and expiratory (E2) neurons, and EPSPs in postinspiratory (PI) neurons. During phase 2 discharge, EPSPs arose in aug-I neurons and IPSPs in PI and E2 neurons. These initial biphasic PSPs were comparable with those during inspiratory off-switching evoked by the same stimulation applied at late-inspiration. In both on- and off-switching, phase-transition in respiratory neuronal activities started to arise concomitantly with the phrenic phase 3 pause. These results suggest that vagal inputs initially produce a non-specific, biphasic response in bulbar respiratory neurons, which consecutively activates a more specific process connected to IonS.     

Responses of ventral respiratory neurones in the rat to vagus stimulation and the functional division of expiration.

In anaesthetized rats, extracellular and intracellular recordings were taken from 106 respiratory neurones in the intermediate region of the nucleus ambiguus. We observed unprovoked shortening of expiratory time accompanied, in all classes of respiratory neurone, by the elimination of the changes in membrane potential that were characteristic of stage II expiration. The demonstration of the elimination of stage II expiration in both the rat and cat strongly supports the functional division of expiration into stage I expiration (post-inspiration) and stage II expiration. In order to identify the neurones in the rat that receive inputs from vagal afferents and modulate the central respiratory rhythm, we examined whether any respiratory neurones responded to stimulation of the vagus nerve. Some post-inspiratory and stage II expiratory neurones responded. The short latency (< 2 ms) of four of the responses indicates that some vagal afferents act on post-inspiratory neurones via a disynaptic pathway. While repetitive stimulation of the vagus nerve could inhibit the phrenic rhythm, it appears that most inspiratory neurones in the intermediate region of the nucleus ambiguous complex are not directly involved in integrating the information from vagal afferents with the central respiratory rhythm.     

Activity patterns in respiratory muscles and in respiratory neurones of the rostral medulla of the cat

1. In the rostral medulla the activity patterns of respiratory neurones are of two main types: inspiratory and expiratory (Fig. 1). Some expiratory neurones begin activity early enough to fit Cohen & Wang's (1959) category of ;inspiratory-expiratory'. However, in the rostral medulla, ;inspiratory-expiratory' neurones do not constitute a distinct category within the expiratory group, since all intermediate activity patterns are observed. Similarly, no distinct categories of activity patterns can be distinguished within the inspiratory group.2. Expiratory neurones are rare in the extreme medial and lateral portions of the respiratory regions on each side of the rostral medulla (Fig. 3). Respiratory neurones are not bunched in anatomical clusters within each region, though a medio-dorsal group of neurones on each side is somewhat anatomically separated from the rest.3. At any one time there are at the very least 1000 respiratory neurones on each side of the rostral medulla.4. Many of the patterns of single unit activity recorded in respiratory muscles of the nose, throat and pharynx are different from the patterns of activity typical of respiratory neurones in the medulla near the cranial motor nuclei (Figs. 1 and 4). This suggests that many of the medullary recordings are from cells other than the motor neurones innervating these muscles.     

Subtype composition and responses of respiratory neurons in the pre-botzinger region to pulmonary afferent inputs in dogs

The brain stem pre-Botzinger complex (pre-BC) plays an important role in respiratory rhythm generation. However, it is not clear what function each subpopulation of neurons in the pre-BC serves. The purpose of the present studies was to identify neuronal subpopulations of the canine pre-BC and to characterize the neuronal responses of subpopulations to experimentally imposed changes in inspiratory (I) and expiratory (E) phase durations. Lung inflations and electrical stimulation of the cervical vagus nerve were used to produce changes in respiratory phase timing via the Hering-Breuer reflex. Multibarrel micropipettes were used to record neuronal activity and for pressure microejection in decerebrate, paralyzed, ventilated dogs. The pre-BC region was functionally identified by eliciting tachypneic phrenic neural responses to localized microejections of DL-homocysteic acid. Antidromic stimulation and spike-triggered averaging techniques were used to identify bulbospinal and cranial motoneurons, respectively. The results indicate that the canine pre-BC region consists of a heterogeneous mixture of propriobulbar I and E neuron subpopulations. The neuronal responses to ipsi-, contra-, and bilateral pulmonary afferent inputs indicated that I and E neurons with decrementing patterns were the only neurons with responses consistently related to phase duration. Late-I neurons were excited, but most other types of I neurons were inhibited or unresponsive. E neurons with augmenting or parabolic discharge patters were inhibited by ipsilateral inputs but excited by contra- and bilateral inputs. Late-E neurons were more frequently encountered and were inhibited by ipsi- and bilateral inputs, but excited by contralateral inputs. The results suggest that only a limited number of neuron subpopulations may be involved in rhythmogenesis, whereas many neuron types may be involved in motor pattern generation.     

Excitatory amino acid-mediated transmission of inspiratory drive to phrenic motoneurons

1. The role of excitatory amino acids (EAAs) in the bulbospinal transmission of inspiratory drive was studied by intracellular and single-electrode voltage-clamp recordings from phrenic motoneurons in the in vitro neonatal rat brain stem spinal cord. 2. In all brain stem-spinal cord preparations there were spontaneously generated rhythmic membrane depolarizations and associated spiking of phrenic motoneurons during the inspiratory phase of the respiratory cycle. The envelope of the motoneuron drive potential had a rapid onset to peak (50 ms) followed by a plateau/declining phase that lasted 400-700 ms. The peak potential was approximately 10-20 mV above base-line potential. The drive current under voltage clamp had a similar shape and duration to the drive potential with a peak current greater than 1.5 nA. 3. The involvement of EAAs in the bulbospinal transmission of inspiratory drive was demonstrated by checking the effects of various EAA receptor antagonists on the phrenic motoneuron inspiratory drive. When kynurenic acid (KYN), an antagonist acting on all three subtypes of EAA receptors, was applied to the solution bathing the spinal cord, the motoneuron action potentials were abolished, and the amplitude of inspiratory drive potential was significantly reduced. To further classify the role of the different EAA receptor subtypes in the synaptic transmission of inspiratory drive, the effects on the drive potential of either 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), a specific non-N-methyl-D-aspartic acid (non-NMDA) receptor antagonist, or DL-2-amino-5-phosphonovaleric acid (AP5), DL-2-amino-7-phosphonoheptanoic acid (AP7), and (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imin emaleate (MK-801), NMDA receptor antagonists, were investigated. Bath or local application of CNQX induced a dose-dependent decrease of the inspiratory drive potential without changing intrinsic motoneuron membrane properties. On the other hand, application of AP7 or MK 801 had a small effect on the inspiratory drive potential or the inspiratory drive current when the motoneuron membrane potential was clamped near end-expiratory potentials (-60 to -75 mV). 4. To establish the presence of EAA receptors on the phrenic motoneuronal membrane and to provide information on the available receptor subtypes for action of the endogenously released transmitter, we tested the effects of agonists for the major EAA receptor subtypes after blocking synaptic transmission (produced by axonal action potentials) by bath application of tetrodotoxin (TTX).(ABSTRACT TRUNCATED AT 400 WORDS)