Postinspiratory neuronal activities during behavioral control, sleep, and wakefulness.

Cells that discharge in early expiration and inhibit other respiratory cells purportedly cause a separate phase of the respiratory cycle that has been named "postinspiration." Our objective was to study these postinspiratory cells in the intact unanesthetized cat during sleep, wakefulness, and behavioral inhibition of inspiration, but we were unable to find cells with strong and consistent activity confined to early expiration. Instead, we found that various cell types were active in early expiration. They included inspiratory-expiratory phase-spanning cells, retrofacial augmenting expiratory cells with bursts in early expiration, retrofacial decrementing expiratory cells, tonic expiratory cells, and cells with variable activity in the early part of expiration. Just as the cell types active during early expiration were heterogeneous so too were their activities during behavioral inhibition of inspiration and during sleep. These results suggest that the state of early expiration is determined by many different cell types rather than a single class of postinspiratory cells.     

Respiratory effects of brief baroreceptor stimuli in the anesthetized dog

To quantify the immediate isocapnic respiratory response to baroreceptor stimulation, pressure in the isolated externally perfused carotid sinuses (CS) of 24 vagotomized alpha-chloralose-anesthetized dogs was increased selectively during either inspiration or expiration as a step (from time of onset to end of respiratory phase) or a pulse (500 ms). The rise time (150 ms), base-line pressure (80 mmHg), and stimulus magnitude (40 mmHg) were similar for the two stimuli. The time of stimulus onset (delay), expressed as a percent of control time of inspiration (TI) or expiration (TE), was varied. TI, TE, and tidal volume (VT) were expressed as percent changes from control. Stimuli delivered early in inspiration lengthened TI [23.5 +/- 6.4% (SE) for step and 11.7 +/- 6.3% for pulse stimuli at 5% delay] more effectively than late stimuli. VT was essentially unaltered. In contrast, step stimuli delivered during expiration caused a lengthening of TE (32.7 +/- 6.3% at 5% delay) that did not depend on the delay (up to 75%). Very late (85%) pulse stimuli lengthened TE (15.2 +/- 5.7%) more effectively than early stimuli. For both stimuli, the expiratory VT was unaltered. When the responses are compared before and after separation of the blood supply of the carotid bodies from the CS region and when they are compared before and after inhibition of reflex systemic hypotension by ganglionic blockade, the observed responses were shown to be due solely to CS baroreceptor stimulation and not to alterations in carotid body blood flow or reflex changes in systemic cardiovascular variables.     

Phase resetting of the respiratory cycle before and after unilateral pontine lesion in cat.

The pontine respiratory group (PRG) facilitates the mechanism for terminating the inspiratory phase but may influence other phases in the respiratory cycle as well. We determined the effects of PRG lesions on the response of the respiratory cycle to superior laryngeal nerve stimulation delivered in each phase of the cycle in decerebrate, vagotomized, paralyzed, and ventilated cats (n = 6). We measured the duration of inspiration (TI) and expiration (TE) for three breaths before and in the perturbed breath and TI for three breaths after the perturbation. The delay to next inspiration was plotted against the phase at which the stimulus was delivered. Before lesioning, premature inspiratory termination was followed by phase-dependent shortening of TE. After lesioning, premature inspiratory termination did not systematically change the following TE. Breath-by-breath variability (measured 50 breaths) increased and stimulus after-effects (prolonged TI in the subsequent cycle) were augmented following lesions. These data indicate that the PRG plays an important role in the control of TE after perturbation and in the stability of the respiratory central pattern generator.     

Activity of bulbar respiratory neurones during cough and other respiratory tract reflexes in cats

Changes evoked by mechanical stimulation of the relevant parts of the respiratory tract in the activity of inspiratory and expiratory neurones in the ventral respiratory group of the medulla oblongata, and in pleural pressure and the diaphragmatic electromyogram, were determined during cough, sneeze and the aspiration and expiration reflexes in 17 anaesthetized (but not paralysed) cats. The results of 72 tests of elicitation of the given reflexes showed that: Compared with the control inspiration, both the mean and the maximum discharge frequency of spontaneously active inspiratory neurones rose during the inspiratory phase of cough, sneeze and the aspiration reflex. Regular recruitment of new inspiratory units was also observed in the inspiratory phase of cough and the aspiration reflex. Compared with the control expiration, both the mean and the maximum discharge frequency of spontaneously active expiratory neurones rose during the cough, sneeze and expiration reflex effort. Recruitment of latent expiratory neurones was always observed in the expulsive phase of the given respiratory processes. The recruitment of latent expiratory neurones was accompanied by reciprocal inhibition of the activity of inspiratory units and recruitment of latent inspiratory neurones by inhibition of the activity of expiratory units and recruitment of latent inspiratory neurones by inhibition of the activity of expiratory units. Regular recruitment of the same expiratory neurones in all expulsive respiratory processes, together with the similar incidence of inspiratory neurones in the inspiratory phase of sneeze and the aspiration reflex, indicates that they are "nonspecific" in character.     

Time-dependent responses of expiration reflex in cats

We investigated the effectiveness of the "expiration reflex" in 10 anesthetized spontaneously breathing cats. The expiration reflex was produced by mechanical stimulation of the vocal folds and electrical stimulation of the superior laryngeal nerve at different moments in the respiratory cycle and at various levels of respiratory chemical drive. The effectiveness of the expiration reflex was evaluated from sudden changes in expiratory flow immediately following the stimulation. Both mechanical and electrical stimulations given during early inspiration caused little or no expiratory efforts, whereas stimulations given during early expiration or hypocapnic apnea produced a typical expiration reflex. Changes in arterial CO2 and O2 partial pressures influenced neither the relationships between the stimulation and its effect on the expiration reflex nor the strength of the expiration reflex. These results indicate that the timing of stimulation with relation to the phase of the respiratory cycle is critical to its effect on the expiration reflex and that changes in respiratory chemical drive do not modify the expiration reflex characteristics.     

Trigeminal reflex regulation of the glottis depends on central glycinergic inhibition in the rat

In an unanesthetized decerebrate in situ arterially perfused brain stem preparation of mature rat, strychnine (0.05-0.2 microM) blockade of glycine receptors caused postinspiratory glottal constriction to occur earlier, shifting from early expiration to inspiration. This resulted in a paradoxical inspiratory-related narrowing of the upper airway. Stimulation of the trigeminal ethmoidal nerve (EN5; 20 Hz, 100 micros, 0.5-2 V) evoked a diving response, which included a reflex apnea, glottal constriction, and bradycardia. After strychnine administration, this pattern was converted to a maintained phrenic nerve discharge and a reduced glottal constriction that was interrupted intermittently by transient abductions. The onset of firing of postinspiratory neurons shifted from early expiration into neural inspiration in the presence of strychnine, but neurons maintained their tonic activation during EN5 stimulation, as observed during control. Inspiratory neurons that were hyperpolarized by EN5 stimulation in control conditions were powerfully excited after loss of glycinergic inhibition. Thus the integrity of glycinergic inhibition within the pontomedullary respiratory network is critical for the coordination of cranial and spinal motor outflows during eupnea but also for protective reflex regulation of the upper airway.     

The interaction between breathing and swallowing in healthy individuals

In this article, we aimed at investigating the interaction between breathing and swallowing patterns in normal subjects. Ten healthy volunteers were included in the study. Diaphragm EMG activity was recorded by a needle electrode inserted into the 7th or 8th intercostal space. Swallowing was monitored by submental EMG activity, and laryngeal vertical movement was recorded by using a movement sensor. A single voluntary swallow was initiated during either the inspiration or expiration phases of respiration, and changes in EMG activity were evaluated. When a swallow coincided with either inspiration or expiration, the duration of the respiratory phase was prolonged. Normal subjects were able to voluntarily swallow during inspiration. During the inspiration phase with swallowing, diaphragmatic activity did not ceased and during the expiration phase with swallowing, there was a muscle activity in the diaphragm muscle.     

Synchronization of respiratory frequency by somatic afferent stimulation.

In cats anesthetized with chloralose-urethan, vagotomized, paralyzed, and artifically ventilated, superficial radial (cutaneous) and hamstring (muscle) nerve afferents were stimulated while phrenic nerve electrical activity was recorded. The results obtained with both types of nerves were similar. Stimulation in mid and late expiration advanced the onset of the next inspiration, shortening its duration. Stimulation in early inspiration advanced, while that in late inspiration delayed, the onset of the next expiration. These effects were often accompanied by changes in phrenic motoneuron firing patterns (earlier recruitment, increased discharge frequency, increased slope of integrated phrenic neurogram). Repetitive somatic afferent stimulation produced sustained increases in respiratory frequency in all cats and in half of them entrainment of respiratory frequency to the frequency of stimulation occurred at ratios such as 4:3, 4:5, 1:2, 1:3, 1:4, and 1:7. The lowest stimulus intensity required for evoking these phase shifts was between 5 and 10T (threshold of most excitable fibers) for muscle afferents and between 1 and 2T for cutaneous afferents. These results demonstrate the existence of a reflex mechanism capable of locking respiratory frequency to that of a periodic somatic afferent input. They also provide an experimental basis for the hypothesis that reflexes are resposible for the observed locking between step or pedal frequency and respiratory rate during exercise in man.     

Analysis of entrainment of respiratory rhythm by somatic afferent stimulation in cats using phase response curves

To elucidate how peripheral somatic afferents synchronize the respiratory rhythm to the exercise rhythm, the phrenic nerve activity in the vagotomized, paralyzed, and artificially ventilated cats anesthetized with chloralose-urethane was recorded during electrical stimulation of the superficial radial nerve afferents. At first, a single pulse train was given at various times of the respiratory cycle to obtain a phase-response curve (PRC). The stimulation given at mid to late expiration produced a phase advance, but the stimulation during inspiration produced no measurable phase shifts in most animals (8/10). The maximum phase advance changed depending on the stimulus intensity. The stronger the stimulus intensity, the greater became the maximum phase advance. Repetitive somatic afferent stimulation produced 1:1 entrainment of the respiratory frequency to the repetitive stimulation. Theoretical predictions on the stable entrainment phase and on the entrainment frequency range from the obtained PRC were close to the experimental results. The present study demonstrated the presence of a neuronal circuit synchronizing the respiratory rhythm to the periodic somatic afferents and the manner of how such entrainment occurs.     

Respiratory neuronal activity during apnea and other breathing patterns induced by laryngeal stimulation

Respiration cycles through three distinct phases (inspiration, postinspiration, and expiration) each having corresponding medullary cells that are excited during one phase and inhibited during the other two. Laryngeal stimulation is known to induce apnea in newborn animals, but the cellular mechanisms underlying this effect are not known. Intracellular recording of ventral respiratory group neurons was accomplished in intact anesthetized, paralyzed, and mechanically ventilated piglets. Apnea was induced by insufflation of the larynx with ammonia-saturated air, smoke, or water. Laryngeal insufflation induced phrenic nerve apnea, stimulation of postinspiratory neurons, and stable membrane potentials in inspiratory and expiratory cells consistent with postinspiratory inhibition. Usually the membrane potential of each neuronal type cycled through an expiratory level before onset of the first recovery breath. Variants of the apnea response, probably reflecting the aspiration reflex or sniffing, sneezing, coughing, and swallowing, were also observed. These latter patterns showed oscillation between inspiration and postinspiration without an apparent intervening stage II expiratory phase. However, stage II expiratory activity always preceded onset of the first ramp inspiration after such a pattern. These findings suggest that activation of postinspiratory mechanisms causes profound alterations in the respiratory pattern and that stage II expiration importantly modulates recovery of ramp inspiratory activity. The mechanism of this latter effect may be inhibition of early inspiratory neurons with consequent postinhibitory rebound.     

Facilitation of the diaphragm response to transcranial magnetic stimulation by increases in human respiratory drive.

The human respiratory neural drive has an automatic component (bulbospinal pathway) and a volitional component (corticospinal pathway). The aim of this study was to assess the effects of a hypercapnia-induced increase in the automatic respiratory drive on the function of the diaphragmatic corticospinal pathway as independently as possible of any other influence. Thirteen healthy volunteers breathed room air and then 5 and 7% hyperoxic CO2. Cervical (cms) and transcranial (tms) magnetic stimulations were performed during early inspiration and expiration. Transdiaphragmatic pressure (Pdi) and surface electromyogram of the diaphragm (DiEMG) and of the abductor pollicis brevis (apbEMG) were recorded in response to cms and tms. During inspiration, Pdi,cms was unaffected by CO2, but Pdi,tms increased significantly with 7% CO2. During expiration, Pdi,cms was significantly reduced by CO2, whereas Pdi,tms was preserved. DiEMG,tms latencies decreased significantly during early inspiration and expiration (air vs. 5% CO2 and air vs. 7% CO2). DiEMG,tms amplitude increased significantly in response to early expiration-tms (air vs. 5% CO2 and air vs. 7% CO2) but not in response to early inspiration-tms. DiEMG,cms latencies and amplitudes were not affected by CO2 whereas 7% CO2 significantly increased the apbEMG,cms latency. The apbEMG,tms vs. apbEMG,cms latency difference was unaffected by CO2. In conclusion, increasing the automatic drive to breathe facilitates the response of the diaphragm to tms, during both inspiration and expiration. This could allow the corticospinal drive to breathe to keep the capacity to modulate respiration in conditions under which the automatic respiratory control is stimulated.     

Transmural pressure in the right atrium during positive pressure respiration in cats

The influence of continuous positive pressure breathing (cm H2O) on the breathing mechanics, central venous pressure, and transmural pressure in the right atrium, were studied in anaesthetised cats separately during inspiration and expiration. It's shown that hemodynamics effects are directly connected with the influence of increased intrathoracic pressure during whole breathing cycles in contrast with the phase changes in natural expiration and inspiration. The inversion of relation of intrathoracic and central venous pressure due to displacement of the mechanical respiratory characteristics became the factors defining the fall of the right atrium filling pressure.     

Integration-differentiation and gating of carotid afferent traffic that shapes the respiratory pattern.

The phase-dependent plasticity of carotid chemoafferent signaling was studied with electrical stimulation of a carotid sinus nerve during either inspiration or expiration in anesthetized, glomectomized, vagotomized, paralyzed, and ventilated rats. Stroboscopic and interferometric analyses of the resulting phase-contrast disturbances of the respiratory rhythm revealed that carotid chemoafferent traffic was dynamically filtered centrally by a parallel bank of leaky integrators and differentiators, each being logically gated to the inspiratory or expiratory phase in a stop-and-go manner as follows: 1) carotid short-term potentiation of inspiratory drive was mediated by dual integrators that both shortened inspiration and augmented phrenic motor output cooperatively in long and short timescales; 2) carotid short-term depression of respiratory frequency was mediated by a (possibly pontine) integrator that lengthened expiration with a relatively long memory; and 3) carotid "chemoreflex" shortening of expiration was mediated by an occult fast integrator, which, together with carotid short-term depression, formed a differentiator. These effects were modulated anteriorly by integrators in the nucleus tractus solitarius that were "auto-gated" to, or recruited by, the carotid sinus nerve input. Such phase-selective and activity-dependent time-frequency filtering of carotid chemoafferent feedback in parallel neurological-neurodynamic central pathways may profoundly affect respiratory stability during hypoxia and sleep and could contribute to the dynamic optimization of the respiratory pattern and maintenance of homeostasis in health and in disease states.     

Gate mechanism in breathlessness caused by chest wall vibration in humans

The effects of bilateral alternating out-of-phase vibrations were studied in 10 normal healthy subjects and five asthmatic patients. The second or third intercostal spaces were vibrated during expiration, and the seventh to ninth intercostal spaces were vibrated during inspiration. Most subjects sensed breathlessness during such vibrations, and 100 Hz was most effective. The degree of breathlessness correlated positively with increased respiratory rate. Respiratory rate increased from 14.1 +/- 3.78 (mean +/- SD) to 22.3 +/- 7.14 breaths/min (P less than 0.05) during relatively severe breathlessness and to 20.39 +/- 5.66 breaths/min (P less than 0.05) during less uncomfortable sensation. Slight or negligible breathlessness induced no significant increase in rate (15.33 +/- 4.19 breaths/min). All asthma patients described the sensations during vibration as similar to those during asthma attacks, and their respiratory rates increased 20.7 +/- 11.03% during 100 Hz vibration (P less than 0.01). It is suggested that the uncomfortable sensation of breathlessness may be induced by muscle spindles in the intercostal muscles being activated out of phase with the respiratory cycle. The central mechanism that receives the intercostal afferents may have a certain gate that operates in relation to the sensation of breathlessness.     

Respiratory effects of stimulation of intercostal muscles and saphenous nerve in kittens

Effects of intercostal muscle stimulation were studied in 2- to 7-day-old kittens under ketamine-acepromazine anesthesia. Animals were vagotomized, paralyzed, and artificially ventilated. Stimuli applied during inspiration (TI) inhibited this phase. Stimulus strength necessary for TI inhibition decreased with time. However, an all-or-nothing effect was not always observed. Stimulation during expiration (TE) prolonged this phase. The responsiveness increased with increasing stimulus delay. The effects of intercostal muscle stimulation were compared with those recorded during saphenous nerve stimulation. Stimulation during TI prolonged this phase. Phrenic activity increased after a short-lasting decrease in the on-going activity. Stimulation during the first 50% of TE had variable effects, whereas stimulation with longer delay shortened this phase. Our results indicated that the pattern of breathing in newborns can be affected by both intercostal muscle and other somatic efferents. However, the mechanisms controlling respiratory timing may differ in newborns and in adults. Different effects of respiratory muscle and saphenous nerve stimulation suggest different transmitters involved or different sites of interaction of these inputs with the medullary respiratory rhythm generator.     

Effects of negative upper airway pressure on pattern of breathing in sleeping infants

Negative upper airway (UAW) pressure inhibits diaphragm inspiratory activity in animals, but there is no direct evidence of this reflex in humans. Also, little is known regarding reflex latency or effects of varying time of stimulation during the breathing cycle. We studied effects of UAW negative pressure on inspiratory airflow and respiratory timing in seven tracheostomized infants during quiet sleep with a face mask and syringe used to produce UAW suction without changing lower airway pressure. Suction trials lasted 2-3 s. During UAW suction, mean and peak inspiratory airflow as well as tidal volume was markedly reduced (16-68%) regardless of whether stimulation occurred in inspiration or expiration. Reflex latency was 42 +/- 3 ms. When suction was applied during inspiration or late expiration, the inspiration and the following expiration were shortened. In contrast, suction applied during midexpiration prolonged expiration and tended to prolong inspiration. The changes in flow, tidal volume, and timing indicate a marked inhibitory effect of UAW suction on thoracic inspiratory muscles. Such a reflex mechanism may function in preventing pharyngeal collapse by inspiratory suction pressure.     

Coordination of swallowing and respiration in unconscious subjects

We investigated the coordination of swallowing and breathing in 11 unconscious patients with an endotracheal tube in place during the recovery period from general anesthesia. Swallows occurred during both the inspiratory and expiratory phases with no preponderant occurrence during either phase. When a swallow occurred during inspiration, the inspiration was interrupted immediately and was followed by expiration, but the durations of both inspiration and expiration were progressively increased as the time from the onset of inspiration to the onset of swallowing was progressively delayed. A swallow coinciding with the expiratory phase progressively prolonged the duration of the expiration that had been interrupted as the timing of swallowing was progressively delayed. Repeated swallows invariably and in a predictable manner caused changes in the breathing pattern. Thus when the frequency of regularly repeated swallows was relatively high, the breathing pattern was characterized by regular, shallow, and rapid breaths. When the frequency of regularly repeated swallows was relatively low, the breathing pattern was characterized by regular, deep, and slow breaths. When the frequency of repeated swallows was irregular, the breathing patterns were characterized by inconsistent changes in tidal volume and respiratory frequency. Our results indicate that, in unconscious subjects, some mechanisms integrating respiration and swallowing are operative and responsible for changes in breathing patterns during swallowing.     

The expiration reflex in the mouse.

The authors determined the elicitability of the expiration reflex and the mechanoreceptive sensitivity of the respiratory tract in 35 anaesthetized albino mice. They found that the expiration reflex could be elicited from both the oral and the tracheal end and that its intensity was statistically significantly higher in elicitation from the oral end. The only other pronounced respiratory reflexes evoked by mechanical stimulation of the airways were the sneeze reflex and the aspiration reflex, which is present in most animals. The mechanoreceptors of the tracheal and bronchial mucosa were less sensitive to stimulation. In most cases they did not react at all, or with just a single forced inspiration, and at other times only by a change in respiration frequency and amplitude.     

Respiratory‐induced vasoconstriction measured by light transmission and by laser Doppler signal

Changes in finger tissue blood volume (TBV) measured by light transmission and in laser Doppler flow (LDF) were obtained during long breathing (of 12 s period) and associated with the respiratory phases, inspiration and expiration. For fifteen out of sixteen subjects TBV and LDF started to decrease 0–2 s after the start of expiration and increased during inspiration but the start of increase occurred before the start of inspiration, showing that the respiratory‐induced changes in TBV and LDF are mainly associated with the expiration. Decrease of TBV and LDF after expiration was also found during the inspiratory gasps (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Respiratory phasic effects of inspiratory loading on left ventricular hemodynamics in vagotomized dogs.

Exaggerated inspiratory swings in intrathoracic pressure have been postulated to increase left ventricular (LV) afterload. These predictions are based on measurements of LV afterload by use of esophageal or lateral pleural pressure. Using direct measurements of pericardial pressure, we reexamined respiratory changes in LV afterload. In 11 anesthetized vagotomized dogs, we measured arterial pressure, LV end-systolic (ES) and end-diastolic transmural (TM) pressures, stroke volume (SV), diastolic left anterior descending blood flow (CBF-D), and coronary resistance. Dogs were studied before and while breathing against an inspiratory threshold load of -20 to -25 cmH2O compared with end expiration. Relative to end expiration, SV and LVES TM pressures decreased during inspiration and increased during early expiration, effects exaggerated during inspiratory loading. In all cases, LV afterload (LVES TM pressure) changed in parallel with SV. LV end-diastolic TM pressure did not change. CBF-D paralleled arterial pressure, and there were no changes in coronary resistance. In two dogs, regional LVES segment length paralleled calculated changes in LVES TM pressure. We conclude that 1) LV afterload decreases during early inspiration and increases during early expiration, changes secondary to those in SV; 2) changes in CBF-D are secondary to changes in perfusion pressure during the respiratory cycle; and 3) the use of esophageal or lateral pleural pressure to estimate LV surface pressure overestimates changes in LV TM pressures during respiration.