Influence of phasic volume feedback on abdominal expiratory nerve activity

Our purpose was to examine the influence of phasic lung volume feedback on the activities of motor nerves innervating the diaphragm and transversus abdominis muscles during hypercapnia and hypoxia. We studied seventeen decerebrate cats that were paralyzed and ventilated with a servo-respirator controlled by the integrated phrenic neurogram. The effects of phasic lung volume feedback were assessed by withholding pulmonary inflation during the central inspiratory period. Withholding lung inflation for a single respiratory cycle under hyperoxic, normocapnic conditions consistently prolonged the durations of the inspiratory and expiratory periods, and caused marked increases in the peak electrical activities of both phrenic and abdominal nerves. Hyperoxic hypercapnia (PaCO2 50-80 mmHg) and isocapnic hypoxia (PaO2 60-35 mmHg) increased peak phrenic and abdominal neural activities, and withholding pulmonary inflation under these conditions caused even greater augmentations of inspiratory and expiratory motor output. The augmentation of expiratory activity by withholding lung inflation was proportionately greater than the concomitant prolongation of the central expiratory period. All responses to non-inflation maneuvers were abolished following bilateral cervical vagotomy. The results indicate that vagally mediated volume feedback during inspiration can attenuate the output of abdominal motoneurons in the subsequent expiratory period. Moreover, hypoxia, which attenuates abdominal motor activity in vagotomized animals, enhances this activity when the vagi are intact.     

Behavior of expiratory neurons in response to mechanical and chemical loading.

The response of medullary expiratory neurons to added mechanical and chemical loads was studied in anesthetized cats. Alterations in burst characteristics and central timing were compared in the intact and bilaterally vagotomized cat. The following results were obtained: (1) Graded expiratory airflow resistances caused progressive increases in burst duration, spikes per burst and firing rate; similar effects were noted for end-inspiratory tracheal occlusions and continuous positive breathing; all facilitation was eliminated by vagotomy. (2) Graded inspiratory airflow resistances delayed the onset of an expiratory burst but did not change the overall burst characteristics. (3) Acute hypercapnia increased ventilation without noticeable changes in expiratory burst characteristics; acute hypoxia produced a reduction in burst duration concomitant with changes in ventilation. It is concluded that (1) expiratory neurons are responsive to vagally mediated volume information and (2) transient hypoxia and hypercapnia sufficient to increase ventilation does not increase the firing rate of expiratory neurons but exerts differential effects with respect to timing. It is suggested that expiratory duration is related to the time integral of expired volume and that the increase in FRC imposed by expiratory loads does not alter the central timing of the next inspiration.     

Respiratory-modulated activities of motor units of the facial nerve

The purpose of this work was to characterize the influence of activity of vagal pulmonary receptors upon the discharge pattern of motor units of the facial nerve. Decerebrate and paralyzed cats were ventilated with a servo-respirator which produced pulmonary inflations in parallel with activity of the phrenic nerve. At normocapnia, facial units discharged phasically during neural inspiration, expiration or across both phases or discharged tonically throughout the respiratory cycle. When pulmonary inflation was withheld, the tonic discharge of some units became phasic; others changed the pattern of phasic discharge. In hypercapnia, the number of tonic fiber activities increased and, again, some phasic discharge patterns were altered. Withholding inflation caused similar alterations as in normocapnia. Activities of facial fibers in vagotomized animals differed in that no tonic activities were recorded, and no change in phasic discharge patterns was induced by hypercapnia. We conclude that afferents from pulmonary stretch receptors influence ventilatory activity throughout the entire respiratory cycle. The concept is discussed that the tonic, as well as phasic discharge of these receptors, is important for the regulation of activity of motoneurons to upper airway muscles.     

Respiratory-associated thalamic activity is related to level of respiratory drive.

We recorded phrenic nerve activity and thalamic single unit firing in unanesthetized, suprathalamically decerebrated, paralyzed and ventilated cats, in which vagi and carotid sinus nerves (CSN) had been ablated. Seventy-six (14%) of 545 neurons in regions of the thalamus related to the ascending reticular system, which had been tonically firing at low respiratory drives, developed rhythmic increases of firing associated with each respiration when drive had been increased by CSN stimulation or hypercapnia. The increases of neuronal firing occurred in late inspiration/post-inspiration but sometimes lasted into expiration; the magnitude of change was graded according to the magnitude of respiratory activity. Thalamic neurons also fired with a rhythm related to ventilator-induced chest expansion, some units showing both the respiratory-associated and the ventilator-related rhythms. Simultaneously recorded mesencephalic and thalamic neurons developed similar rhythms when drive was increased. We suggest that these neuronal activities reflect the conveyance of information about respiration to the cortex, where it may lead to the sensation of dyspnea and perhaps to arousal.     

The effects of hypercapnia and hypoxia on single hypoglossal nerve fiber activity

The respiratory related modulation of hypoglossal nerve activity has been studied at the single fiber level in cats under hyperoxic hypercapnia and hypoxic conditions and their conduction velocities determined. Changes in fiber activity were compared to simultaneous changes occurring in phrenic activity. Three different kinds of discharge patterns were observed: (a) inspiratory, (b) phasic activity during both inspiration and expiration, and (c) continuous random activity with no respiratory modulation. These fibers could be grouped into three categories according to their pattern of discharge during CO2 breathing. Type I fibers, mean conduction velocity of 30.0 m/sec, exhibited only an inspiratory phasic discharge during 100% O2 breathing. Their discharge frequency increased rapidly with higher levels of CO2 and hypoxia. Type II fibers, mean conduction velocity of 36.7 m/sec, had three different kinds of inspiratory-expiratory discharge patterns during 100% O2 breathing. With increasing hypercapnia or hypoxia fibers of this group discharged phasically during inspiration and discharge at low frequency during expiration. Type III fibers had a non phasic discharge pattern at 100% O2 breathing and at all levels of CO2 tested (up to 10%). Discharge frequency rose during CO2 rebreathing and hypoxia, but the rate of increase was much less than Type I and Type II fibers. Their mean conduction velocity was 41.3 m/sec. The inspiratory activity of Type I and II fibers increased their activity more than the phrenic during hypercapnia and hypoxia. Type II and Type III fibers are responsible at least in part for the tonic activity of the nerve.     

Differential effects of hypercapnia on expiratory phases of respiration in the piglet

Hypercapnia induces prolongation of expiratory time (TE) during early development. In the present study, we determined the response to steady state hypercapnia of three neural phases of the total respiratory cycle, inspiration (TI), stage 1 or passive expiration, TE-1 and stage 2 or active expiration, TE-2. Experiments were performed in decerebrate, vagotomized, spontaneously breathing piglets aged 5-10 days. Neural phases of the respiratory cycle were based on electrical activities of the thyroarytenoid (TA, laryngeal adductor) and triangularis sternii (TS, chest wall expiratory muscle) in relation to diaphragm (D) activity. We observed that hypercapnia induced prolongation of both expiratory phases. The greater prolongation of TE-1 was associated with an increase in TA activity and an increase in laryngeal resistance, which peaked early in TE-1, and then progressively decreased. These findings demonstrate that, in early postnatal life, a hypercapnia induced increase in respiratory drive is associated with centrally mediated prolongation of both phases of expiration, a greater prolongation of TE-1, and an increase in laryngeal resistance during post-inspiration. We speculate that the latter serves to optimize gas exchange by reducing large fluctuations in functional residual capacity.     

Activity of the respiratory muscles during natural defecation: a study on experimental animals]

Activity of the respiratory muscles during natural defecation was studied in two anesthetized and two decerebrate dogs. In anesthetized dogs, excitation of the abdominal muscles and an increase in gastric pressure were observed during defecation. However, pleural pressure was little influenced by such increase in abdominal pressure, maintaining the same rhythmic changes as observed during spontaneous respiration. The rhythmic changes in pleural pressure were associated with rhythmic activity of the diaphragm. When gastric pressure increased during defecation, the diaphragmatic activity also increased during both the inspiratory and expiratory phases. In a decerebrate dog, airflow and airway pressure changed similarly to during defecation. The diaphragm was continuously active, with superimposed rhythmic augmentation. In a paralyzed and artificially ventilated dog with open-chest, the phrenic nerve similarly developed discharges. We conclude that the non-respiratory activity and rhythmic augmentation of phrenic nerve discharge during defecation is pre-programmed in the command for defecation. The activity of phrenic motoneurons may be further modulated by changes in thoracic and abdominal pressure. These mechanisms may act together to coordinate respiration and defecation.     

Responses of recurrent laryngeal motoneurons to changes of pulmonary afferent inputs

In decerebrate, paralyzed cats ventilated with a cycle-triggered pump, the discharges of the recurrent laryngeal (whole nerve or single fibers) and phrenic nerves, and the changes produced by pulmonary afferent inputs (lung inflation), were compared. When lung inflation was in phase with neural inspiration, four types of laryngeal fiber activities were observed: (a) phasic-inspiratory; (b) tonic-inspiratory; (c) expiratory-inspiratory; (d) early-expiratory. The firing patterns during inspiration were plateau-like, whereas the phrenic pattern was augmenting. When inflation was withheld, the plateau patterns usually became augmenting, indicating inhibition of laryngeal inspiratory activity by pulmonary afferents. Secondary effects of withholding inflation were (a) increases of early-expiratory activity (both whole nerve and individual fiber), indicating increased post-inhibitory rebound excitation; (b) decreased activity of tonic-inspiratory and expiratory-inspiratory fibers during early neural expiration, indicating increased inhibition by early-expiratory neurons. The discharge patterns of different types of laryngeal motoneuron, as well as their changes with inflation, are interpreted in relation to the function of regulating airway resistance.     

The effect of antigen-induced bronchoconstriction on phrenic nerve activity

The effects of acute bronchoconstriction, produced by inhalation of Ascaris suum antigen, on both the amplitude and timing of phrenic nerve activity were studied in anesthetized dogs. Blood gas tensions and inspiratory air flow were maintained constant. Bronchoconstriction resulted in a significant increase in the magnitude and rate of rise of the phrenic neurogram during both normal respiratory cycles and cycles when lung inflation was prevented. The increase in the slope of the phrenic neurogram that results from lung inflation was further increased during bronchoconstriction. In addition, in half the animals, there was a significant increase in the tonic phrenic activity measured during expiration. All of these changes were abolished by bilateral cervical vagotomy. Antigen administration did not affect the timing of the different phases of the cycle equally; inspiratory duration was reduced by 51.8% and expiratory duration by 67.6%. Postinspiratory activity of the phrenic (PIIA) was reduced by only 34.7%; thus, during bronchoconstriction PIIA occupied a proportionally greater fraction of the expiratory phase. Vagotomy abolished the changes in respiratory timing and eliminated all PIIA.     

CO2-induced prolongation of expiratory time during early development.

In these studies, we determined the contribution of central mechanisms and the role of GABA(A)-receptor signal transduction pathways in mediating hypercapnia-induced slowing of breathing frequency. Experiments were performed in decerebrate, vagotomized, paralyzed and mechanically ventilated piglets of 3-5 days and 2-3 weeks of age (n=19). Repeated exposure to progressive hyperoxic hypercapnia induced a reproducible increase in phrenic nerve activity, accompanied by a CO2 concentration-dependent increase in expiratory duration. No differences were observed in piglets with intact or cut carotid sinus nerves. Intravenous administration of bicuculline (2 mg/kg: n=7), a gamma-aminobutyric acid (GABA(A)) receptor antagonist, significantly reduced the CO2-induced prolongation of TE. These data demonstrate for the first time that in early postnatal life, hypercapnia induced increase in phrenic activity is associated with centrally mediated prolongation of expiratory duration. Furthermore. the results suggest that brainstem GABAergic mechanisms play an important role in CO2-induced prolongation of expiratory time during early development.     

Functions of the retrofacial nucleus in chemosensitivity and ventilatory neurogenesis

The hypothesis was evaluated that neurons within the retrofacial nucleus of medulla integrate afferent stimuli from the central chemoreceptors. In decerebrate, vagotomized, paralyzed and ventilated cats, activity of the phrenic nerve was monitored. Peak integrated phrenic activity increased in hypercapnia; the frequency of phrenic bursts typically declined slightly. The retrofacial nucleus was ablated by radio-frequency lesions or neurons within this nucleus were destroyed by microinjections of kainic acid. Results were similar following lesions or injections. Following unilateral ablations, peak phrenic activity was greatly reduced at normocapnia and hypercapnia; the frequency of phrenic bursts typically rose. Both frequency and peak phrenic activity fell further after the contralateral destruction with a cessation of all phasic phrenic discharge being observed in most animals. Injections of kainic acid in regions rostral, caudal or medial to the retrofacial nucleus produced no consistent changes in phrenic activity. We conclude that neuronal activities in the region of the retrofacial nucleus are important both in the integration of stimuli from the central chemoreceptors and in defining the discharge patterns of respiratory neurons.     

Respiratory changes in abdominal configuration in supine dogs

In an attempt to understand the respiratory changes in abdominal muscle length in supine dogs (Ninane et al., 1988), we have recorded the electromyographic (EMG) activity of the transversus abdominis, external oblique, and rectus abdominis in eight supine, lightly anesthetized animals, and we have measured the respiratory changes in anteroposterior (AP) and transverse (T) diameters of the abdomen. Five animals had phasic expiratory EMG activity in the transversus during room air breathing, while only two animals had expiratory activity in the external oblique; no animal had phasic expiratory activity in the rectus. Activation of the transversus during expiration was invariably associated with a decrease in the abdominal T diameter and a rise in gastric pressure. In contrast, the abdominal AP diameter tended to increase. These alterations in abdominal configuration remained unchanged after denervation of the triangularis sterni, but decreased in magnitude when activation of the transversus was reduced by supplemental anesthesia. Conversely, these alterations in abdominal configuration increased in magnitude when expiratory activation of the transversus was increased by hyperoxic hypercapnia. These observations indicate that in supine dogs: (1) Expiratory contraction of the transversus acts primarily to reduce the transverse diameter of the abdomen; (2) This reduction, in turn, promotes an increase in abdominal pressure which results secondarily in an outward motion of the ventral abdominal wall; and (3) The latter may explain why the rectus abdominis, although electrically silent, shortens during expiration below its in situ relaxation length. The present observations also establish that in supine dogs breathing at rest, the abdomen does not move with a single degree of freedom.     

Stimulating fastigial nucleus alters central mechanisms regulating phrenic activity

In paralyzed, mechanically ventilated, anesthetized cats, 30-sec trains of electrical stimulation in the rostral fastigial nucleus caused either respiratory excitation or inhibition (apnea) of respiration. The response elicited by stimulation was dependent on the frequency of stimulation: the duration of apnea decreased and respiratory excitation occurred more frequently at lower frequencies of stimulation. Excitation was characterized by increases of f and the rate of rise and peak (TNA) integrated activity of the phrenic nerve. TI and TE decreased. TNA and f remained elevated for at least three minutes after excitatory respiratory responses. Also, respiration was frequently elevated after an inhibitory response. Brief stimulation (100-200 ms) administered during expiration or inspiration altered either TE or TI and TNA, respectively. In addition, brief stimulation elicited short-latency inhibition or excitation of phrenic nerve activity. These effects were often unassociated with other phase changes. We conclude that activation of neurons or axons within the rostral fastigial nucleus can modulate activity of the phrenic nerve by altering the activity of at least three separate central mechanisms.     

Comparison of respiratory-related trigeminal, hypoglossal and phrenic activities

In decerebrate, paralyzed and vagotomized cats, we recorded activities of hypoglossal and phrenic nerves and of the mylohyoid branch of the trigeminal nerve. At normocapnia, a respiratory-modulated trigeminal discharge could be discerned in most cats. This discharge was characterized by a diminution of activity during neural inspiration and a peak in expiration. In hypercapnia or hypoxia, peak activity increased and its time of occurrence moved to late inspiration. Augmentations of peak trigeminal, hypoglossal and phrenic activities were proportional. Peak trigeminal and hypoglossal activities increased more than phrenic following administrations of protriptyline, strychnine and, in some cats, cyanide or doxapram. Peak trigeminal activity fell more than phrenic after diazepam. Pentobarbital or halothane reduced peak hypoglossal, but not trigeminal, activity more than phrenic. However, after these anesthetics, trigeminal activity became restricted to the inspiratory-expiratory junction. We conclude that trigeminal and hypoglossal activities are more dependent upon processes within the reticular formation than is the bulbospinal-phrenic system. Central and peripheral chemoreceptor influences are distributed equivalently upon trigeminal, hypoglossal and phrenic motoneurons.     

Role of the medullary raphe nuclei in the respiratory response to CO2

We characterized the role of neurons within the midline of the medulla oblongata on phrenic and hypoglossal nerve responses to hypercapnia during early development. Studies were performed on decorticate or anesthetized, vagotomized and mechanically ventilated 14–20 day old piglets. Reversible withdrawal of midline neuronal activity was induced by microinjections of lidocaine (2%, 300 nl; n=10) and lesioning was caused by microinjections of the neurotoxic agent, ibotenic acid (n=12), at the same sites. At any given end-tidal CO₂, peak phrenic and hypoglossal activities after lidocaine were significantly lower than in the control period (P<0.01). Similarly, 1–2 h after injections of ibotenic acid, both phrenic and hypoglossal nerve responses to CO₂ were significantly lower than in the control period (P<0.01). The results indicate for the first time that the medullary midline neurons are required for full expression of ventilatory responses to hypercapnia and raise the possibility that dysfunction of these nuclei may contribute to respiratory instability during early postnatal life.     

Long term facilitation of phrenic motor output

Episodic hypoxia or electrical stimulation of carotid chemoafferent neurons elicits a sustained, serotonin-dependent augmentation of respiratory motor output known as long term facilitation (LTF). The primary objectives of this paper are to provide an updated review of the literature pertaining to LTF, to investigate the influence of selected variables on LTF via meta-analysis of a large data set from LTF experiments on anesthetized rats, and to propose an updated mechanism of LTF. LTF has been demonstrated in anesthetized and awake experimental preparations, and can be evoked in some human subjects during sleep. The mechanism underlying LTF requires episodic chemoafferent stimulation, and is not elicited by similar cumulative durations of sustained hypoxia. Meta-analysis of phrenic nerve responses following episodic hypoxia in 63 experiments on anesthetized rats (conducted by four investigators over a period of several years) indicates that phrenic LTF magnitude correlates with peak phrenic responses during hypoxia and hypercapnia, but not with the level of hypoxia during episodic exposures. Potential mechanisms underlying these relationships are discussed, and currently available data are synthesized into an updated mechanistic model of LTF. In this model, we propose that LTF arises predominantly from episodic activation of serotonergic receptors on phrenic motoneurons, activating intracellular kinases and, thus, phosphorylating and potentiating ionic currents associated with the glutamate receptors that mediate respiratory drive.     

Comparison of activities of medullary respiratory neurons in eupnea and apneusis

We evaluated patterns of antidromic latencies of medullary respiratory neurons in eupnea and apneusis to define how afferent influences from the pneumotaxic center regulate their activities. Apneusis was reversibly produced by cold block in decerebrate, vagotomized, paralyzed and ventilated cats. Most neurons, which discharged during all of eupneic inspiration or expiration, maintained the same pattern in apneusis. However, those active during only portions of these phases or spanning both changed markedly with alterations in periods of discharge, including tonic patterns or cessations of activity. Such marked changes were observed for activities of all laryngeal expiratory neurons. Upon termination of eupneic discharge, most bulbospinal and laryngeal neurons had transient peaks of latencies, indicating hyperpolarizations; declines from these peak values were greatly reduced in apneusis. Moreover, reflecting depolarizations, latencies of some inspiratory and expiratory neurons declined during eupneic expiration and inspiration, respectively; these declines were much reduced in apneusis. We conclude that the pneumotaxic center influences medullary respiratory neuronal activities not only at end-inspiration, but throughout the entire respiratory cycle.     

Physiological role of respiratory-related bronchial constriction]

The respiration-related rhythmic constriction of the fifth-generation bronchi was analyzed in 11 tracheostomized dogs. During spontaneous breathing, the bronchial pressure (Pbr) estimated with a balloon-tipped catheter increased almost in parallel with the pleural pressure (Ppl) in the early inspiratory phase, but decreased in the late inspiratory phase. The parallel duration/inspiratory duration was 0.72 +/- 0.19 (mean +/- SD). This finding was more prominent in hypercapnia, but statistical significance was not obtained. When the efferent phrenic nerve fibers were electrically stimulated (pulse train, 0.1 ms, 30 Hz, 5 V, 2 s), Pbr changed almost in parallel with Ppl during the inspiratory phase, and expiration was completed significantly earlier than during spontaneous breathing (time constant 0.17 +/- 0.06 s vs 0.26 +/- 0.07 s). Bronchial constriction in early expiration may increase airway pressure and keep patency of the peripheral bronchi.     

Responses of early and late onset phrenic motoneurons to lung inflation

In anesthetized or decerebrate cats that were paralyzed and ventilated with a cycle-triggered pump, we produced changes in activity of the whole phrenic nerve and of individual phrenic motoneurons (fibers or cells in the spinal cord) by withholding lung inflation during the inspiratory (I) phase. The neurons were classified into early- and late-onset types (discharge onset less or greater than 80 msec, respectively, after whole phrenic onset). Both unit and whole phrenic activity exhibited a variety of responses to inflation (excitation, depression, or no effect); but there were no consistent differences between responses of early- and late-onset neurons. The distribution of responses was quite different from that of dorsal respiratory group (DRG) I neurons (Cohen and Feldman, 1984); in particular there was no group of phrenic neurons corresponding to the late-onset I-beta neurons (I neurons excited by inflation). We conclude that the inputs to late-onset phrenic neurons are not predominantly or exclusively from late-onset DRG neurons.     

Properties of postganglionic sympathetic neurons with axons in phrenic nerve.

The aim of the study was to test the reflex and resting properties of postganglionic sympathetic neurons with axons located in the right phrenic nerve. The experiments have been performed on chloralose-anesthetized cats with both vago-aortic nerves cut. The somata or the postganglionic sympathetic neurons were located in the stellate ganglion. Axons of these neurons passed through the upper and lower phrenic nerve roots and through the phrenic nerve itself. The presence of cardiac and respiratory rhythmicities was detected in the activity of the phrenic postganglionic sympathetic neurons. Hyperventilation, which abolished burst discharges of the phrenic nerve, decreased the sympathetic activity by 14%. Systemic hypoxia (ventilating the animals for 2 min with 8% O2 in N2) increased the sympathetic activity threefold. The results of our experiments suggest that axons of the sympathetic neurons located in the right phrenic nerve could possibly be diaphragmatic muscle vasoconstrictors.