Inspiratory on-switch evoked by mesencephalic stimulation: activity of medullary respiratory neurones

Summary The activity of medullary inspiratory and expiratory neurones was studied in urethan-chloralose anaesthetized cats during stimulus — evoked inspiratory phase (inspiratory on-switch). All neurones were characterized according to their axonal destination (i.e. bulbospinal neurones or vagal motoneurones) or the absence of such axonal projections (i.e. propriobulbar neurones), and to their location in the dorsal or ventral respiratory nuclei. 1. The inspiratory on-switch effects were elicited during expiration (E phase) by brief tetanic electrical stimulation (50 to 100 ms duration; 0.5 mA; 300 Hz) delivered to the mesencephalic periaqueductal central gray and the adjacent reticular formation. The evoked inspiratory effects observed on the phrenic nerve discharge consisted of: (i) an immediate response (latency 20 ± 5 ms) of stable duration related to the stimulus (primary response: Prim.R.), (ii) a delayed response (patterned response: Patt.R.) appearing after a latent period (silent phase: Sil.P.) of 100 ms maximal duration. The later the stimulus in the E phase, the longer was the duration of the Patt.R. (300 to 1000 ms). 2. The stimulation evoked an earlier activation of the inspiratory bulbospinal neurones (latency 12 ± 6 ms) than that obtained in the phrenic nerve (Prim.R.). Hence, the Prim.R. originated from the bulbospinal pathway and not from a pathway directly impinging on the motoneurones. Conversely during stimulation very few inspiratory propriobulbar neurones were activated and no expiratory neurone discharged. 3. During the phrenic Sil.P., 46% of the inspiratory bulbospinal neurones continued to discharge with a firing rate lower than that during the stimulus train, while most of the inspiratory propriobulbar and expiratory neurones were not active. 4. During the Patt.R. all inspiratory bulbospinal neurones discharged early and were strongly activated whatever the Patt.R. duration whereas the expiratory neurones were not active. Inspiratory propriobulbar neurones were either not recruited or recruited later, and the number of active neurones increased as the duration of the Patt.R. lengthened. 5. Our results suggest that the eliciting of the stimulus-evoked inspiration (Patt.R.) primarily depends on the activation of the inspiratory bulbospinal neurones. These neurones therefore would not only be the output neurones of the medullary respiratory centres, but they would serve other roles such as building up of the excitation in other respiratory neurones, thus acting as a component of the inspiratory ramp generator.Abbreviations Prim.R Primary response - Patt.R Patterned response - Sil.P Silent phase - I phase Inspiratory phase - E phase Expiratory phase - IBSN Inspiratory bulbospinal neurones - IPBN Inspiratory propriobulbar neurones - EBSN Expiratory bulbospinal neurones - EPBN Expiratory propriobulbar neurones - DRN Dorsal respiratory nucleus - VRN Ventral respiratory nucleus Supported by CNRS (LA 205 and ATP no 4188) and Fondation pour Ia recherche médicale     

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.     

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     

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.     

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.     

Interaction of pulmonary afferents and pneumotaxic center in control of respiratory pattern in cats.

The interaction between the pulmonary afferents (PA) and the pneumotaxic center (PC) in control of respiratory pattern was studied in lightly anesthetized paralyzed cats before and after bivagotomy or lesions of the PC using inflations controlled by the onset or cessation of phrenic nerve discharge, i.e., cycle-triggered inflations. This interaction was also studied using electrical stimulation of the central stumps of cut vagi. Introduction of a delay between inspiratory onset and the commencement of an inflation at constant flow and duration resulted in increases of the durations of inspiration (T1) and expiration (TE) and amplitude of the integrated phrenic nerve discharge (A). The lung volume at inspiratory cutoff, i.e., the volume threshold, increased markedly as T1 increased. There were linear relationships between T1 and TE and between T1 and A. At constant alveolar CO2 and tidal volume, the quantitative effects of delay were dependent on the rate of inflation; i.e., when the flow increased, the volume threshold for a given T1 decreased. Bilateral vagotomy abolished the effects of delay and flow. PC lesions, which resulted in apneusis when the cycle-triggered inflations were stopped, produced the following changes compared to the delay effects seen in intact cats: a) the volume threshold for zero delay doubled and its rate of decrease with increased T1 was significantly smaller, and b) the change in TE for a given change in T1 was reduced markedly. Introduction of a delay between inspiratory onset and the start of electrical stimulation of the afferent vagi resulted in effects similar to those seen for delays in cycle-triggered inflations. The T1-TE relationship remained linear when the stimulus trains ended with inspiratory cessation. These results suggest that: a) the inspiratory cutoff mechanism is responsive to the rate, as well as the level, of lung inflation; b) all of the lung volume information affecting inspiratory cutoff in paralyzed cats is carried via the vagi; c) an intact PC is necessary for the generation of a normal time dependence of the volume threshold for inspiratory cutoff; d) the PC plays an important role in matching TE to T1 when the latter changes. For inflations and vagal stimulations applied during expiration, with introduction of a delay between inspiratory cessation and the start of cycle-triggered inflation or vagal stimulation, the results indicated that the expiratory cutoff mechanism has an irrevocable phase of 300-450 ms.     

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     

Respiratory depression caused by either morphine microinjection or repetitive electrical stimulation in the region of the nucleus parabrachialis of cats

In chloralose-urethane anesthetized, vagotomized, paralyzed and artificially ventilated cats, respiratory response to either repetitive electrical stimulation or micro-injection of morphine in the rostral pons was studied by recording the phrenic nerve discharges. In the region of the nucleus parabrachialis (PBN) and its ventral reticular formation, electrical stimulation delivered in 20 successive expiratory periods caused the respiratory depression to last long after the termination of stimulation. This respiratory-depressant effect could be reversed by naloxone. By a single electrical stimulation delivered in most of these effective sites, a phasic phrenic excitation was consistently elicited in the period of both expiration and inspiration, and the reduction in expiratory duration could be observed when the stimulation was delivered in expiratory period. In the microinjection study of 2.66 nmol morphine in 0.1 l in the localized area of the dorsolateral portion of the PBN, a significant reduction in both respiratory outputs and the rate of increase in inspiratory activity could be induced within 1 min after the application. The respiratory depression thus caused by both methods was quite similar in several respiratory variables. Thus an involvement of the PBN region in long-lasting respiratory modulation mediated by endogenous opioid system is suggested.     

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.     

D1-dopamine receptor blockade slows respiratory rhythm and enhances opioid-mediated depression

Previous studies indicate that dopamine modulates the excitability of the respiratory network and its susceptibility to depression by exogenous opioids, but the roles of different subtypes of dopamine receptor in these processes are still uncertain. In this study, D1-dopamine receptor (D1R) involvement in dopaminergic modulation of respiratory rhythm and mu-opioid receptor mediated depression were investigated in pentobarbital-anesthetized cats. Intravenous administration of the D1R blocker SCH-23390 (100-200 microg/kg) slowed phrenic nerve and expiratory neuron respiratory rhythms by prolonging the inspiratory and expiratory phases. Phrenic nerve discharge intensity also increased more gradually during the inspiratory phase. SCH-23390 (150 microg/kg) also enhanced dose-dependent depression of phrenic nerve and expiratory neuron excitability, as well as rhythm disturbances, produced by the mu-opioid receptor agonist fentanyl (2-20 microg/kg, i.v.). The results suggest an important role for the D1-subtype of receptor in respiratory rhythm modulation, and indicate that this type of receptor participates in dopaminergic compensatory mechanisms directed against opioid-mediated network depression.     

Augmentation of muscle nociceptive respiratory reflex facilitation by vagal afferents

Vagal influence on the facilitation of phrenic neural activity during respiratory phase-locked, gastrocnemius muscle nerve nociceptive electrical stimulation was examined in anesthetized, glomectomized, paralyzed, and artificially ventilated cats. (1) In the vagi-intact state, respiratory reflex facilitation was characterized by a sharp rise in peak amplitude, maximum rate of rise or slope, and mean rate of rise of integrated phrenic nerve activity. This was greater during inspiratory phase-locked (T1-locked) muscle nerve electrical stimulation than during expiratory phase-locked (TE-locked) muscle nerve electrical stimulation. "Evoked post-inspiratory phrenic activity" during the early expiratory phase was also observed during TE-locked muscle nerve electrical stimulation. (2) Bilateral vagotomy significantly attenuated the respiratory facilitation during both T1- and TE-locked muscle nerve electrical stimulation. In particular, the "evoked post-inspiratory phrenic activity" during TE-locked muscle nerve electrical stimulation was also attenuated or almost completely abolished. (3) Conditioning electrical stimulation of the vagus nerve revealed facilitatory reflexes which co-exist with inspiratory inhibitory reflexes. (4) The "evoked post-inspiratory phrenic activity" during TE-locked muscle nerve electrical stimulation, which was attenuated or abolished after vagotomy, was restored after vagal T1-locked conditioning stimuli combined with TE-locked muscle nerve electrical stimulation. The results suggest that vagal facilitatory reflexes augment the respiratory reflex facilitation during muscle nociceptive stimulation.     

Modification by chemical stimuli of temporal difference in the onset of inspiratory activity between vagal (superior laryngeal) or hypoglossal and phrenic nerves of the rat

Temporal differences in the onset of inspiratory activities between the efferent vagal (superior laryngeal, Xsl) or hypoglossal (XII) and phrenic (Phr) nerves were measured at various levels of chemical stimuli in the halothane-anesthetized, vagotomized, and artificially ventilated rat. The onset of Xsl (XII) inspiratory activities always preceded the abrupt start of the Phr discharge. Hyperoxic hypocapnia due to hyperventilation delayed the start of inspiratory activity (reduction in respiratory frequency) and shortened the difference in onset time between the cranial (Xsl, XII) and Phr nerve discharges (Td). During respiratory stimulation due to asphyxia (progressive hypoxia and hypercapnia), the start of Xsl (XII) inspiratory activity became progressively earlier than that of Phr discharge, which extremely prolonged the Td. Severe asphyxia, however, retarded the start of inspiratory activities with accompanying long Td and slow respiratory frequency. The early but gradually augmenting inspiratory activity of the Xsl (XII) nerve was always followed by large bursts synchronized with Phr discharges during altered chemical stimuli. The termination of inspiratory activity, which occurred simultaneously in the three respiratory nerves, was not significantly affected by changes in chemical stimuli except for extreme hypocapnia. The results indicate that changes in chemical stimuli not only alter the start of inspiratory activity but also influence the transition from the initial slow onset to the final synchronized inspiratory activity in the Xsl (XII) nerve. The apparent dissociation of the onset time between the Xsl (XII) and Phr nerve discharges shows that the temporal aspect of the brain stem process(es) for starting inspiratory activities may not be determined from the trajectory of Phr discharges only.     

Possible involvement of the facial nucleus in regulation of respiration in rats

The facial nucleus (FN) has been known as a motor nucleus to control the activity of the facial skeletal muscles by its efferent somatic motoneurons. Much less, however, is known about the non-motor control functions of its interneurons. The present study was designed to investigate if the interneurons of the FN participate in controlling rhythmic respiration in the sodium thiopental-anesthetized and vagotomized Sprague-Dawley rats with facial motoneurons retrogradely degenerated with techniques of electrical and chemical stimulation of the FN and extracellular recording of discharge of neurons in the FN. Single pulse stimulation (75-100 microA, 0.2 ms) of the FN during inspiration caused a transient restrain in phrenic discharge. Short train stimulation (75-100 microA, 0.2 ms, 100 Hz, 3-5 pulses) delivered during the early- or mid-term of inspiration augmented the inspiratory duration, but switched the inspiration off when delivered during the later stage of inspiration. Short train stimulation delivered during expiration prolonged the expiratory duration. Continuous stimulation could inhibit the inspiration. Microinjection of kainic acid into the FN caused an augmentation in inspiratory duration and amplitude and in expiratory duration. These data indicate that the interneurons of the FN might participate in the modulation of respiration. Different discharge patterns of interneurons in the FN, interestingly some respiratory related patterns, were observed, which provide a possible structural basis for the role of the FN in respiratory regulation.     

Respiratory interneurons of the lower cervical (C4-C5) cord: membrane potential changes during fictive coughing,vomiting, and swallowing in the decerebrate cat

The possible roles of interneurons in the C4-C5 cervical spinal cord in conveying central drives to phrenic motoneurons during different behaviour patterns were investigated using intracellular recordings in decerebrate, paralysed, artificially ventilated cats. Eleven cells were tentatively classified as respiratory interneurons since they: (i) could not be antidromically activated from the ipsilateral whole intrathoracic phrenic nerve, and (ii) exhibited large membrane potential changes during eupnea (7.3 mV±3.6, range 2–13.5 mV) or non-respiratory behaviour patterns. Six neurons depolarized in phase with phrenic discharge; four others depolarized during the expiratory phase; one neuron exhibited depolarization during the end of both expiration and inspiration. A variety of responses was observed during fictive coughing, vomiting, and swallowing. The results are consistent with C4-C5 expiratory interneurons conveying inhibition to phrenic motoneurons during different behaviour patterns. The responses of inspiratory and multiphasic neurons suggest that the roles of these interneurons are mode complex than simply relaying central excitatory or inhibitory drive to phrenic motoneurons.     

Extra-segmental reflexes derived from intercostal afferents: phrenic and laryngeal responses

1. Phrenic and recurrent laryngeal efferent responses were evoked by brief tetani or single shocks to the cut external intercostal nerves of anaesthetized cats. The reflexes derived from middle thoracic segments (T5 and 6) were compared with those emanating from caudal thoracic segments (T9 and 10).2. During inspiration, middle intercostal nerve stimulation transiently inhibited the spontaneous discharge in both efferent neurograms, whereas stimulation of caudal intercostal nerves facilitated phrenic discharge and usually inhibited recurrent laryngeal activity.3. During expiration, stimulation at either thoracic level enhanced recurrent laryngeal discharge while provoking little or no phrenic response.4. Superficial lesions of the lateral cervical cord, ipsilateral to the stimulus sites, above or below the phrenic outflow, eliminated all reflex responses except the phrenic response to caudal thoracic stimuli. Similarly, in the spinal animal, middle intercostal afferents could not be shown to decrease phrenic excitability. Caudal intercostal afferents cause phrenic excitation by a spinal reflex.5. Group I afferents of the mid-thoracic segments and group II afferents of the caudal thoracic segments initiate these extra-segmental reflexes.6. The recurrent laryngeal responses manifest, for the most part, changes in the discharge of fibres innervating the posterior cricoarytenoid muscle. The responses fit the overall pattern of response to middle intercostal nerve stimulation, namely, inhibition of inspiratory muscles and excitation of expiratory muscles. Intercostal afferent stimulation also activated the laryngeal adductor muscles.7. The results support the view that intercostal mechanoreceptors initiate an array of extra-segmental respiratory reflexes, including spinal and supraspinal arcs. The simplest way to account for the various responses to stimulation of middle intercostal afferents is to postulate a reflex involving supraspinal respiratory neurones.8. The observed reflexogenic differences correlate with anatomical differences between the middle and caudal ribs. Possible functional implications of this relationship are discussed.     

The importance of timing on the respiratory effects of intermittent carotid sinus nerve stimulation

1. The respiratory response, measured directly as tidal volume or indirectly by using integrated peak phrenic activity, to intermittent electrical stimulation of the carotid sinus nerve was determined in anaesthetized cats.2. Stimulation at rates of 20-25 Hz for 0.5 sec had a rapid effect, increasing inspiratory airflow and phrenic discharge, but only if applied during inspiration. An increase in tidal volume or peak level of integrated phrenic discharge occurred only if the stimulus was exhibited during the second half of inspiration. Continuous stimulation had no greater effect on size or frequency of breathing than did intermittent inspiratory stimuli alone. Stimulation during expiration had no effect on the form or magnitude of subsequent breaths.3. Stimuli in expiration led to a prolongation of expiration. Stimuli in late inspiration caused a prolongation of both inspiration and expiration. Because of these effects, the respiratory rate could be changed by stimulation; in some instances entrainment of respiration by the intermittent carotid sinus nerve stimuli occurred.4. The findings are attributable to modulation of incoming carotid sinus nerve information by the central respiratory neurones, which use primarily that which arrives during inspiration. They show a possible mechanism by which oscillating signals may have a different effect than their mean level would indicate.     

Effects of lesions in the parabrachial nucleus on the mechanisms for central and reflex termination of inspiration in the cat.

The effects of PCO2 and body temperature on the time course and peak amplitude of the central inspiratory activity (CIA) and the inspiratory "off-switch" threshold was studied in apneustic and non-apneustic cats. Apneusis resulted from lesions of the inspiratory inhibiting structures of the medial parabrachial nucleus (NPBM) and by interrupting vagal volume feedback. The cats were paralyzed and ventilated either proportionally to their phrenic output or at predetermined rate and volume. The dependence of the rate of rise and maximal amplitude of phrenic activity on PCO2 and body temperature were comparable in apneustic and non-lesioned animals. The Hering-Breuer volume threshold for inspiratory termination was increased following the rostral pontine lesions. Both hyperthermia and hypercapnia caused augmentation of the absolute rate of rise of inspiratory activity but hypercapnia, in contrast to hyperthermia, caused virtually no change in the fractional increment per unit time. With hypercapnia the inspiratory "off-switch" threshold was raised in the apneustic animals in intact ones, whereas hyperthermia did not seem to influence this threshold. In apneustic conditions expiratory duration remained constant, independent of the large variations in the inspiratory durations. Our results suggest that the NPBM merely provides an excitatory, threshold-lowering input to the inspiratory "off-switch" mechanism.     

The bulbar network of respiratory neurons during apneusis induced by a blockade of NMDA receptors

Summary Our aim was to study the mechanisms producing the transition from the inspiratory phase to the expiratory phase of the breathing cycle. For this purpose we observed the changes affecting the discharge patterns and excitabilities of the different types of respiratory neurons within the respiratory network in cat medulla, after inducing an apneustic respiration with the N-methyl-D-aspartate (NMDA) antagonist MK-801 given systemically. Respiratory neurons were recorded extracellularly through the central barrel of multibarrelled electrodes, in the ventral respiratory area of pentobarbital-anesthetized, vagotomized, paralyzed and ventilated cats. Inhibitions exerted on each neuron by the presynaptic pools of respiratory neurons were revealed when the neuron was depolarized by an iontophoretic application of the excitatory amino-acid analogue quisqualate. Cycle-triggered time histograms of the spontaneous and quisqualate-increased discharge of respiratory neurons were constructed in eupnea and in apneusis induced with MK-801. During apneustic breathing, the activity of the respiratory neuronal network changed throughout the entire respiratory cycle including the post-inspiratory phase, and the peak discharge rates of all types of respiratory neurons, except the late-expiratory type, decreased. During apneusis, the activity of the post-inspiratory neuronal pool, the post-inspiratory depression of other respiratory neurons, and the phrenic nerve after-discharge were reduced (but not totally suppressed), whereas the discharge of some post-inspiratory neurons shifted into the apneustic plateau. The shortened post-inspiration (stage 1 of expiration) altered the organization of the expiratory phase. Late-expiratory neurons (stage 2 of expiration) discharged earlier in expiration and their discharge rate increased. The inspiratory on-switching was functionally unaffected. Early inspiratory neurons of the decrementing type retained a decrementing pattern followed by a reduced discharge rate in the apneustic plateau, whereas early-inspiratory neurons of the constant type maintained a high discharge rate throughout the apneustic plateau. Inspiratory augmenting neurons, late-inspiratory and offswitch neurons also discharged throughout the apneustic plateau. During the apneustic plateau, the level of activity was constant in the phrenic nerve and in inspiratory neurons of the early-constant, augmenting, and late types. However, progressive changes in the activity of other neuronal types demonstrated the evolving state of the respiratory network in the plateau phase. There was a slowed but continued decrease of the activity of early-inspiratory decrementing neurons, accompanied by an increasing activity and/or excitability of off-switch, postinspiratory and late-expiratory neurons. In apneusis there was a decoupling of the duration of inspiration and expiration. The variability of inspiratory duration increased five-fold whereas the variability of expiration was unchanged. We conclude that in the apneustic state, (1) inspiratory on-switching and the successive activation of the different inspiratory neuronal types are preserved; (2) near the end of the inspiratory ramp, the reversible phase of inspiratory off-switching is prolonged, producing the apneustic plateau, and (3) the irreversible phase of offswitching is impaired by a reduced activity of postinspiratory neurons. These results support the 3-phase model of respiratory rhythm generation, in which key roles are played by early-inspiratory and post-inspiratory neurons.     

Role of the ventrolateral region of the nucleus of the tractus solitarius in processing respiratory afferent input from vagus and superior laryngeal nerves

Summary The role of respiratory neurons located within and adjacent to the region of the ventrolateral nucleus of the tractus solitarius (vlNTS) in processing respiratory related afferent input from the vagus and superior laryngeal nerves was examined. Responses in phrenic neural discharge to electrical stimulation of the cervical vagus or superior laryngeal nerve afferents were determined before and after lesioning the vlNTS region. Studies were conducted on anesthetized, vagotomized, paralyzed and artificially ventilated cats. Arrays of 2 to 4 tungsten microelectrodes were used to record neuronal activity and for lesioning. Constant current lesions were made in the vlNTS region where respiratory neuronal discharges were recorded. The region of the vlNTS was probed with the microelectrodes and lesions made until no further respiratory related neuronal discharge could be recorded. The size and placement of lesions was determined in subsequent microscopic examination of 50 m thick sections. Prior to making lesions, electrical stimulation of the superior laryngeal nerve (4–100 A, 10 Hz, 0.1 ms pulse duration) elicited a short latency increase in discharge of phrenic motoneurons, primarily contralateral to the stimulated nerve. This was followed by a bilateral decrease in phrenic nerve discharge and, at higher currents, a longer latency increase in discharge. Stimulation of the vagus nerve at intensities chosen to selectively activate pulmonary stretch receptor afferent fibers produced a stimulus (current) dependent shortening of inspiratory duration. Responses were compared between measurements made immediately before and immediately after each lesion so that changes in response efficacy due to lesions per se could be distinguished from other factors, such as slight changes in the level of anesthesia over the several hours necessary in some cases to complete the lesions. Neither uni- nor bi-lateral lesions altered the efficacy with which stimulation of the vagus nerve shortened inspiratory duration. The short latency excitation of the phrenic motoneurons due to stimulation of the superior laryngeal nerve was severely attenuated by unilateral lesions of the vlNTS region ipsilateral to the stimulated nerve. Neither the bilateral inhibition nor the longer latency excitation due to superior laryngeal nerve stimulation was reduced by uni- or bi-lateral lesions of the vlNTS region. These results demonstrate that extensive destruction of the region of the vlNTS: a) does not markedly affect the inspiratory terminating reflex associated with electrical stimulation of the vagus nerve in a current range selective for activation of pulmonary stretch receptor afferents, and b) abolishes the short-latency increase, but not the bilateral decrease or longer latency increase in phrenic motoneuronal discharge which follows stimulation of the superior laryngeal nerve. We conclude that respiratory neurons in the region of the vlNTS do not play an obligatory role in the respiratory phase transitions in this experimental preparation. Neurons in the vlNTS region may participate in other reflexes, such as the generation of augmented phrenic motoneuronal discharge in response to activation of certain superior laryngeal or vagus nerve afferents.     

Functional associations among simultaneously monitored lateral medullary respiratory neurons in the cat. I. Evidence for excitatory and inhibitory actions of inspiratory neurons

Data were obtained from 45 anesthetized (Dial), paralyzed, artificially ventilated, bilaterally vagotomized cats. Arrays of extracellular electrodes were used to monitor simultaneously the activities of lateral medullary respiratory neurons located in the rostral and caudal regions of the ventral respiratory group. The average discharge rate as a function of time in the respiratory cycle was determined for each neuron and concurrent phrenic nerve activity. Most cells were tested for axonal projections to the spinal cord or the ipsilateral vagus nerve using antidromic stimulation techniques. Seven hundred and sixty-one pairs of ipsilateral respiratory neurons that contained at least one neuron whose maximum discharge rate occurred during the inspiratory phase were analyzed by cross-correlation of the simultaneously recorded spike trains. Twenty-three percent of the 410 pairs of inspiratory (I) neurons showed short time scale correlations indicative of functional association due to paucisynaptic connections or shared inputs. Eight per cent of the 351 pairs composed of an I cell and and expiratory (E) neuron were correlated. We found evidence for excitation of both bulbospinal I neurons and I cells that were not antidromically activated by stimulation of the spinal cord and vagus nerve (NAA neurons) by NAA I cells. We also obtained data suggesting inhibitory actions of cells whose maximum discharge rate occurred in the first half of the I phase (I-DEC neurons). These actions included inhibition of other I-DEC neurons, inhibition of cells whose greatest firing rate occurred in the last half of the I phase (I-AUG neurons), inhibition of E-DEC neurons, and inhibition of E-AUG cells. Sixty-two percent (31/50) of the correlations that could be interpreted as evidence for an excitatory or inhibitory paucisynaptic connection were detected in pairs composed of a caudal and a rostral ventral respiratory group neuron. Eighty-eight percent (14/16) of proposed intergroup excitatory connections involved a projection from the rostral neuron of the pair to the caudal cell, whereas 73% (11/15) of proposed inhibitory connections involved a caudal-to-rostral projection. These results support and suggest several hypotheses for mechanisms that may help to control the development of augmenting activity in and the timing of each phase of the respiratory cycle.