Respiratory activity in the superior laryngeal nerve of the rabbit

We studied the respiratory modulation of laryngeal afferents and their response to transmural pressure in 24 anesthetized, spontaneously breathing rabbits. Laryngeal afferent activity has a predominant inspiratory augmentation during tracheal breathing or tracheal occlusion that can be accounted for by the respiratory movement transmitted to the larynx through the trachea. During upper airway breathing or upper airway occlusion SLN afferent activity increases in expiration and decreases in inspiration. This respiratory modulation is due to changes in upper airway pressure (Pua). In fact, positive pressure stimulates SLN afferent activity, while negative pressure inhibits it. Mechanical restriction of epiglottal movement reduced the response to Pua changes during upper airway occlusion and application of maintained positive (0.1-0.5 kPa) and negative (-0.1 to -0.5 kPa) pressures (P less than 0.005). Furthermore, surgical removal of epiglottis decreased the baseline activity of SLN to 16.5% of control. These experiments suggest that in the rabbit the epiglottis is the main source of SLN afferent activity and that its displacement, due to changes in Pua, is the most important factor for modulating SLN activity. Most of the laryngeal receptors showed an inspiratory augmentation with tracheal breathing and occlusion, were stimulated by positive pressure and inhibited by negative pressure, reflecting the behavior observed in the whole nerve.     

The respiratory activity of the superior laryngeal nerve in the rat.

The aim of this study was to characterize the laryngeal afferent activity of the rat. The animals were anesthetized and breathing spontaneously. Laryngeal afferent activity was recorded from both the whole superior laryngeal nerve (SLN) and from single fibers isolated from this nerve. An overall inspiratory augmenting activity was observed in the whole SLN during tracheostomy breathing, tracheal occlusion and upper airway breathing, but an expiratory augmenting activity was present during upper airway occlusion. The inspiratory modulated activity was abolished by bilateral section of the hypoglossal nerves but not the recurrent laryngeal nerves. A great number of receptors (46/80, 58%) were identified as 'drive' receptors, and others as 'pressure' (22/80, 28%) and 'irritant' type receptors (9/80, 11%). Nineteen pressure receptors were stimulated by positive transmural pressure, while only three stimulated by negative pressure. Nine drive receptors were also stimulated by positive pressure and inhibited by negative pressure. Such response to pressure was further evaluated by applying maintained pressures to the functionally isolated upper airway. These results are essentially consistent with findings obtained in the rabbit, but differ from those reported for the dog.     

Sulphur dioxide block of laryngeal receptors in rabbits

In 6 rabbits moving average of activity of superior laryngeal nerve(SLN) increased when pressure in upper airways (Pua) was positive and decreased when it was negative. After SO2 exposure of upper airways SLN activity at Pua = 0 decreased to 40% and was no longer affected by changes in Pua. Activity of 67 fibers of SLN was recorded in 11 rabbits: 35 came from 'pressure' receptors, 27 from 'drive', and 5 from 'flow'. Thirty-three pressure receptors discharged at Pua = 0: 32 increased their firing rate with positive Pua and decreased it with negative Pua, one did the reverse. One pressure receptor silent at Pua = 0 fired with positive Pua, the other with negative Pua. Pressure receptors were slowly adapting. SO2 blocked within 3-9 min 84% of pressure receptors, 56% of drive receptors, and 4 out of 5 flow receptors. The receptors recovered control activity within 5-10 min after SO2 removal. SO2 block of laryngeal receptors may represent a convenient experimental tool for studies of laryngeal reflexes.     

Reflex partitioning of inputs from stretch receptors of bronchi and thoracic trachea

A selective block of slowly adapting stretch receptors in anesthetized rabbits was induced by exposing to SO2 all thoracic airways (T) or the carina and bronchi alone (B). Increment of inspiratory time (TI) relative to control was 61% greater under B than T. The reverse would have happened if input responsible for Breuer-Hering inflation reflex originated from both bronchi and trachea. Hence, bronchial input activates inspiratory off-switch, while tracheal input delays its activation. During single inspiratory efforts with airways closed at end expiration diaphragm activity (Adi) decreased and TI0 increased relative to control equally under B and T. Hence, the input facilitating central inspiratory activity at end expiratory volume does not stem from trachea. At end of inspiratory ramp Adi stopped within 43 msec at control and 57 msec under B and T. Hence, bronchial input speeds up off-switching of inspiration. Postinspiratory Adi was greater under B than T, and nearly nil at control. Hence, bronchial input inhibits postinspiratory Adi, while tracheal input facilitates it. Inspiratory and expiratory flows were more damped under B than T, and under T than at control.     

Influence of negative pressure applied to the upper airway on the breathing pattern in unanesthetized awake dogs

We examined the influence of changes in upper airway pressure on the breathing pattern in 5 unanesthetized awake dogs. The dogs breathed through an endotracheal tube or through a comfortably fitting fiberglass snout mask. With matched resistances and volume of the dead space, the inspiratory duration, tidal volume, and minute ventilation were higher during nasal breathing compared to tracheal breathing. Nasal and tracheal occlusion produced prolongation of inspiration in the first occluded breathing attempt, but the prolongation was more marked in nasal occlusion tests. Augmentation of genioglossus muscle activity occurred on the first occluded breath in nasal but not tracheal occlusion. In another series of experiments, negative pressure was applied to the isolated upper airway while the dog breathed through a tracheostomy tube. Negative pressure caused a prolongation of inspiratory duration which was proportional to the level of the applied pressure. However, the prolongation of inspiratory duration was significantly more marked when application of negative pressure was timed simultaneously with tracheal occlusion. Our results demonstrate that the upper airway has a powerful effect on the control of breathing, which becomes more evident during tracheal occlusion.     

Respiratory afferent activity in the superior laryngeal nerves

This study evaluates the afferent activity in the superior laryngeal nerve (SLN) during breathing as well as during occluded inspiratory efforts. Experiments were performed in 11 anesthetized and spontaneously breathing dogs. Electroneurographic activity was recorded from the peripheral cut end of the SLN and, in 3 dogs, also from the contralateral vagus nerve. A tracheal cannula with a side arm allowed the bypass of the larynx during breathing and occluded efforts. A clear inspiratory modulation was present in all experimental conditions. Both peak and duration of the SLN activity decreased (87% and 89%) when breathing was diverted from the upper airway to the tracheostomy. Peak and duration of the SLN activity (as % of upper airway breathing) increased during occluded efforts; however, the increase was greater when the larynx was not by-passed (peak = 118% vs 208%, duration = 143% vs 178%). Section of the ipsilateral recurrent laryngeal nerve reduced the inspiratory modulation. Vagal afferent activity increased equally during tracheostomy and upper airway breathing and decreased markedly during tracheal and upper airway occlusions. Our results indicate that collapsing pressure in the larynx is the major stimulus in activating laryngeal afferents.     

Ventilation in kittens with chronic section of the superior laryngeal nerves

The breathing pattern of conscious newborn kittens one-to-two weeks old was studied by the barometric method about 5 days after bilateral section of the superior laryngeal nerve (SLN-denervated group) or a sham operation (SLN-sham operated group). None of the ventilatory variables differed between the two groups, whether during normoxia or acute hypoxia (10 min of 10% O2). After anesthesia, delivery of steady airflows in the expiratory direction through the upper airways of the SLN-sham operated had marked inhibitory effects on ventilation which entirely disappeared after SLN section. A small inhibition was still present in the SLN-denervated group, possibly indicating that other non-SLN upper airways receptors developed inhibitory ventilatory effects during the period of chronic denervation. Intermittent expiratory upper airway airflows were much less effective than steady flows and no inhibition was seen with oscillatory flows, indicating that the mode of application of the stimulus to the laryngeal receptors is crucial in determining the magnitude of their reflex response. Under anesthesia, acute bilateral section of the SLN determined a small increase of the integrated peak EMG activity of the diaphragm. We conclude that laryngeal SLN afferents are inhibitory on ventilation in newborn kittens, but this effect is very small during normal conscious conditions. Only under special circumstances, including anesthesia and sustained upper airways flows and pressures, the ventilatory inhibition can be disproportionately magnified.     

Breathing pattern in rats with chronic section of the superior laryngeal nerves

In a first set of experiments we registered the integrated afferent activity of the superior laryngeal nerve (SLN) in adult anaesthetized rats. The activity increased with positive upper airway pressure (Pua); with progressively more negative Pua, the SLN activity at first declined then increased again. A second set of adult rats underwent bilateral section of the SLN (SLN denervated) or a sham operation (controls). Both groups appeared to recover promptly from the operation and 6 days later their resting breathing pattern was recorded by the barometric method. SLN denervated rats had a shorter inspiratory time (80%), hence higher frequency and mean inspiratory flow, than controls. During hypoxia (10 min at 10% O2) both groups hyperventilated with an almost identical pattern. The rats were then again anaesthetized and the right vagus cut in an attempt to reduce the afferent component from the lower airways, which may have masked the SLN regulatory contribution. One week after this second operation both SLN denervated and controls breathed more deeply and slowly than before vagotomy, but the pattern was not significantly different between the two groups, either in normoxia or hypoxia. Finally, the rats were anaesthetized and integrated diaphragm activity recorded during spontaneous breathing and the first effort against closure of the nostrils. With both vagi cut, the duration of the occluded effort was slightly longer in SLN denervated than in controls. These results suggest that in adult awake rats laryngeal afferent activity tends to decrease mean inspiratory flow. However, this regulatory contribution is small during eupnea and insignificant during hypoxic hyperventilation.     

Reflex influences from the extrathoracic trachea during airway occlusion.

We have performed experiments in 26 dogs anesthetized with pentobarbital and fitted with an endotracheal tube. The inflatable cuff of this tube was positioned either at the level of the cricoid cartilage or at the thoracic inlet. In this latter situation the extrathoracic trachea (E.T.T.) is not subjected to any change in transmural pressure both during breathing and airway occlusion. We have compared the inspiratory output in term of the integrated phrenic discharge during airway occlusion at FRC with the tracheal tube positioned at either one of the two levels. In most of the experiments (16 out of 26) the inspiratory output during airway occlusion is significantly greater (157%) when the E.T.T. is not by-passed and this difference disappears after bilateral vagotomy. We interpret these results by the asymmetrical response of the tracheal stretch receptors to positive and negative transmural pressure (Pt); most of these receptors are active at FRC and decrease their activity at low negative Pt, as that attained in the first occluded breath. These results seem to suggest that the reflex influences from the extrathoracic tracheal receptors on the inspiratory output are similar to those originating from the intrathoracic airway stretch receptors.     

Role of intrinsic muscles and tracheal motion in modulating laryngeal receptors

Recording from the superior laryngeal nerve discloses a respiratory modulated activity even in the absence of airflow and pressure changes in the larynx. The present study evaluates the relative contribution of intrinsic laryngeal muscle activity and transmitted tracheal movement on the respiratory modulation of laryngeal mechanoreceptors. Seventy-four receptors were studied in 22 anesthetized spontaneously breathing dogs. The modulation of 31 receptors depended solely on laryngeal muscle activity since it was abolished by cold block of laryngeal nerves. Twelve receptors were primarily activated by tracheal movement since tracheal stabilization alone reduced or abolished their modulation. The respiratory modulation of the remaining 31 receptors was found to be dependent on both laryngeal muscle activity and tracheal movements. Lidocaine (2%) was applied to the receptor field of 13 endings; the results indicate that while some receptors are located superficially (blocked within 1 min) others are located in deeper structures (not affected in 30 min). These receptors may be involved in the precise coordination of laryngeal muscle activity and could play a role in the regulation of breathing pattern and airway patency due to their pressure sensitivity.     

Effects of cooling on laryngeal reflexes in the dog

We investigated the reflex effects of laryngeal cooling on posterior cricoarytenoid (PCA) muscle activity, breathing pattern, arterial blood pressure and heart rate. We performed experiments on 9 anesthetized, spontaneously breathing dogs. Laryngeal temperature was decreased by passing cold air through the functionally isolated larynx while the dog was breathing through a tracheostomy. Inspiratory and expiratory durations, esophageal pressure, peak PCA activity, heart rate and blood pressure did not change significantly during laryngeal cooling. Upon interruption of cold airflow, while the laryngeal temperature was returning to control values, we assessed PCA response to upper airway occlusion. At laryngeal temperatures of 20-25 degrees C the peak PCA activity during upper airway occlusion was approximately 2/3 of that observed at control temperature (approximately equal to 33 degrees C). This difference was abolished by topically applied anesthetics or by superior laryngeal nerve section. In addition, we recorded from 4 laryngeal mechanoreceptors stimulated by negative pressure; their response to upper airway occlusion was reduced to 1/2 by laryngeal cooling. These results indicate that laryngeal cooling has a marked depressive effect on the PCA response to collapsing pressure in the larynx, thereby compromising the mechanism subserving upper airway patency.     

Role of airway mechanoreceptors in the inhibition of inspiration during mechanical ventilation in humans

The purpose of this study was to demonstrate a neuromechanical inhibitory effect on respiratory muscle activity during mechanical ventilation and to determine whether upper and lower airway receptors provide this inhibitory feedback. Several protocols were completed during mechanical ventilation: (1) positive and negative pressure changes in the upper airway, (2) airway anesthesia to examine the consequences of receptor blockade on respiratory muscle activity, (3) increasing FRC with positive end-expiratory pressure to study the effect of hyperinflation or stretch on respiratory muscle activity, and (4) use of heart-lung transplant patients to determine the effects of vagal denervation on respiratory muscle activity. All subjects were mechanically hyperventilated with positive pressure until inspiratory muscle activity was undetectable and the end-tidal PCO2 decreased to less than 30 mm Hg. End-tidal PCO2 (PETCO2) was increased by either adding CO2 to the inspired gas or decreasing tidal volume (50 ml/min). The PETCO2 where a change in inspiratory muscle activity occurred was taken as the recruitment threshold (PCO2RT). Neuromechanical feedback caused significant inspiratory muscle inhibition during mechanical ventilation, as evidenced by the difference between PCO2RT and PETCO2 during spontaneous eupnea (45 +/- 4 versus 39 +/- 4 mm Hg) and a lower PCO2RT when tidal volume was reduced with a constant frequency and fraction of inspired CO2. Recruitment threshold was unchanged during positive and negative pressure ventilation, during upper and lower airway anesthesia, and in vagally denervated lung transplant patients. These findings demonstrate that neuromechanical feedback causes highly significant inhibition of inspiratory muscle activity during mechanical ventilation; upper and lower airway receptors do not appear to mediate this effect.     

Effect of nasal obstruction on upper airway muscle activation in normal subjects

It is not known whether nasal occlusion produces obstructive sleep apnea (OSA) by decreasing upper airway muscle activation via nasal reflexes or by increasing upper airway resistance and hence lowering the pressure in the pharnyx. The purpose of this study was to determine the effect of nasal occlusion on upper airway muscle activation. We studied seven men and measured alae nasi (AN) and genioglossal (GG) electromyograms (EMGs) during two nights of sleep, one with their nose open and the other with their nose occluded. Nasal occlusion produced OSA in all subjects and also increased the percentage of time during sleep in which phasic AN and GG EMG activity was present. Apneas tended to occur at the nadirs of EMG activity. This suggests that nasal occlusion generally increases respiratory drive to upper airway muscles during sleep and that it does not cause OSA by merely decreasing respiratory drive to these muscles.     

Effects of halothane,enflurane, and isoflurane on laryngeal receptors in dogs

The effects of halothane, enflurane, and isoflurane on laryngeal receptors were investigated in 6 anethetized dogs breathing spontaneously through a tracheostomy. Single unit actiion potentials were recorded from the peripheral cut end of the superior laryngeal nerve (SLN) while different concentrations of volatile anesthetics (1.25, 2.5., 5.0%) were administered in the expiratory direction at a constant air-flow (6 1/min) for 1 min through the functionally isolated upper airway. A total of 21 respiratory-modulated mechanoreceptors, 18 “irritant” receptors, and 7 cold receptors were studied. The overall results obtained from the 16 respiratory-modulated mechanoreceptors challenged with the 3 anesthetic gases disclosed a prevalent inhibitory effect and halothane proved to be the most effective of the 3 gases. The activity during both the inspiratory and expiratory phase was significantly reduced only by halothane (inspiratory phase, P<0.01; expiratory phase, P<0.05), while neither isoflurane nor enflurane caused significant changes in receptor activity. Of the 18 irritant receptors, 14 receptors increased their activity in a dose-related manner in response to one or more of the anesthetics although the effect of halothane was more pronounced than those of enflurane and isoflurane. All of the 7 cold receptos consistently increased their activity in a dose-related manner in response to halothane whereas 3 of 7 receptors were insensitive to enflurane and 4 of 7 receptors were insensitive to isoflurane. Our results indicate that, while all three commonly used anethetics can have an effect on different types of laryngeal receptors, the effects of halothane are more pronounced than those of the other two gases in terms of changes in receptor activity.     

Effect of upper airway negative pressure on respiratory timing

The effects of upper airway negative pressure on respiratory timing and respiratory muscle activity were investigated in 13 urethane-pentobarbital anesthetized adult rabbits. Diaphragm and upper airway muscle EMGs were recorded with fine wire electrodes. The upper airway was converted into a closed system and negative pressure changes were made at will with a syringe attached to a laryngeal cannula. Both inspiratory and expiratory durations (Ti and Te) were prolonged during the negative pressure trials. Maximal prolongation occurred on the first experimental breath for Te and on second breath for Ti. Decreased effects were seen during maintained negative pressures. Peak diaphragm EMG and average slope of diaphragm EMG decreased during these trials. Diaphragmatic apnea (Te greater than or equal to 5 sec) occurred in 15% of trials. In some of these trials apnea lasted as long as the negative pressure stimulus whereas in others spontaneous breathing resumed after a period of apnea. Phasic upper airway muscle activity occurred during diaphragmatic apnea in most of these trials. The superior laryngeal nerve section markedly reduced the effects of negative pressure, indicating that its afferents primarily mediate this response. Our results suggest that upper airway negative pressure acts centrally on both inspiratory and expiratory timing as well as on the motor output of thoracic and upper airway respiratory muscles.     

Laryngeal receptors responding to transmural pressure, airflow and local muscle activity

The larynx has a rich sensory supply which is the main source of several respiratory reflexes. These reflexes, that influence both the patency of the upper airway and the pattern of breathing, are related to transmural pressure and/or airflow in the upper airway. Yet hardly any information is available on the response of laryngeal mechanoreceptors to transmural pressure and airflow. We recorded action potentials from single fibers separated from the superior laryngeal nerve of anesthetized dogs, breathing spontaneously either through a tracheostomy or the upper airway. The airway could be occluded above or below the larynx. On the basis of their behavior during tracheostomy breathing, upper airway breathing, tracheal occlusion and upper airway occlusion, laryngeal mechanoreceptors were classified as pressure receptors, flow receptors or 'drive' receptors (stimulated by the respiratory activity of upper airway muscles). Pressure receptors were encountered most frequently, representing 63.6% of our sample of 110 receptors, 'drive' receptors constituted 21.8% and flow receptors the remaining 14.6%. Our findings indicate that, even though the three types of receptors differ in sensory modality, they concur in exhibiting a predominant activity during inspiration. In fact, 65% of all receptors are active during eupneic inspiration. Moreover, their activity increases markedly during upper airway obstruction.     

Influence of the upper airway on breathing pattern and expiratory time constant in unanesthetized dog pups

Unanesthetized dog pups (2 to 31 days old) respond to sudden opening of a tracheal cannula to atmospheric pressure with a marked increase in breathing frequency. This response is achieved with a 25% decrease in inspiratory and 40% decrease in expiratory times. Expiratory thyroarytenoid muscle activity increased concomitantly, while inspiratory diaphragmatic and posterior cricoarytenoid muscle activities were reduced. These responses are interpreted as a compensatory mechanism for maintenance of an elevated end-expiratory lung volume with functional loss of the upper airway. The changes in expiratory time and thyroarytenoid muscle activity were not observed when positive pressure was applied at the trachea. The expiratory time constant was assessed during spontaneous breathing. The mean value was twice as long during nasal breathing than during tracheal breathing. The nasal value was substantially increased when the thyroarytenoid muscle was active during expiration.     

Inspiratory effort sensation to added resistive loading in patients with obstructive sleep apnea

STUDY OBJECTIVES: Repeated episodes of upper-airway occlusion are the main characteristics of patients with obstructive sleep apnea (OSA) during sleep. It has been reported that an impairment in the sensation of detection and a depression of ventilatory compensation to added load could be observed in such patients. In this study, we examined patients with OSA to evaluate the inspiratory effort sensation (IES), ventilation, and mouth occlusion pressures during added resistive loading while awake and to determine whether they can be reversed by nasal continuous positive airway pressure (CPAP) treatment. DESIGN: A hospital-based case-control study. SETTING: A sleep laboratory of a medical unit in Japan. SUBJECTS: Seventeen patients with moderate to severe OSA and 10 control subjects were included in this study. MEASUREMENTS: All patients with OSA had undergone standard nocturnal polysomnography. Patients with OSA and control subjects were evaluated for IES measured by a modified Borg score, ventilation, and mouth occlusion pressure during control and inspiratory resistive loaded breathing. These tests were repeated in all patients with OSA after 2 weeks of nasal CPAP treatment. RESULTS: IES to inspiratory resistive loading was lower in patients with OSA than in control subjects. There were no differences in ventilation and mouth occlusion pressure between patients and control subjects during loaded breathing. After 2 weeks of nasal CPAP, the decreased IES was increased in patients with OSA. CONCLUSION: In patients with OSA, the decreased IES to inspiratory resistive loaded breathing is reversible with nasal CPAP. This could be one additional benefit of nasal CPAP in the treatment of OSA.     

Effect of laryngeal anesthesia on pulmonary function testing in normal subjects

Pulmonary function tests (PFT) were performed on 11 normal subjects before and after topical anesthesia of the larynx. The PFT consisted of flow volume loops and body box determinations of functional residual capacity and airway resistance, each performed in triplicate. After the first set of tests, cotton pledgets soaked in 4% lidocaine were held in the pyriform sinuses for 2 min to block the superior laryngeal nerves. In addition, 1.5 ml of 10% cocaine was dropped on the vocal cords via indirect laryngoscopy. PFT were repeated 5 min after anesthesia. Besides routine analysis of the flow volume loops, areas under the inspiratory (Area I) and expiratory (Area E) portions of the loops were calculated by planimetry. Area I, peak inspiratory flow (PIF), as well as forced inspiratory flow at 25, 50, and 75% forced vital capacity (FVC), decreased after anesthesia. Peak expiratory flow decreased after anesthesia, but Area E and forced expiratory flow at 25, 50, and 75% FVC were unchanged. This protocol also was performed in 12 normal subjects with isotonic saline being substituted for the lidocaine and cocaine. In this group, no significant differences were observed when flow volume loop parameters were compared before and after topical application of saline. In 5 spontaneously breathing anesthetized dogs, posterior cricoarytenoid muscle and afferent superior laryngeal nerve activity were recorded before and after laryngeal anesthesia performed with the same procedure used in the human subjects. Laryngeal anesthesia resulted in a substantial decrease or a complete disappearance of afferent SLN activity recorded during unobstructed and obstructed respiration. The data suggest that laryngeal receptors help modulate upper airway patency in man.     

Effects of stretch receptors of bronchi or trachea on genioglossus muscle activity

Activity of genioglossus muscle (GG) was recorded in anesthetized rabbits at control and under SO2 block of slowly adapting stretch receptors in thoracic airways (T) or in bronchi alone (B). At control peak activity occurred at 10% of inspiratory time, followed by a slight and a marked decrease. T and, particularly, B delayed onset and slowed rise of GG activity; then, this increased slightly under T and more under B, becoming greater than at control. During inspiratory efforts at end-expiratory volume GG activity at control was equal to that during open inspirations up to its peak, then progressively greater; during inspiratory efforts GG activity under T and B was equally smaller than at control. These findings indicate that bronchial input facilitates GG activity at end-expiratory volume and inhibits it at larger volumes: these effects are greater than those previously detected on diaphragm. Moreover, tracheal input inhibits onset and development of GG activity. Under all conditions end of inspiratory activity was simultaneous in GG and in diaphragm. Time to peak inspiratory flow correlated with time to early peak in GG activity.