F.B. Sant'Ambrogio

SENSORY INFORMATION FROM THE UPPER AIRWAY: ROLE IN THE CONTROL OF BREATHING

The functional integrity of extrathoracic airways critically depends on the proper orchestration of the activities of a set of patency-maintaining muscles. Recruitment and control of these muscles is regulated by a laryngeal and trigeminal affects that originate from pressure sensing endings. These sensors are particularly numerous among laryngeal receptors and, indeed, they constitute the main element in the respiration-modulated activity of the superior laryngeal nerve. Considering that the most compliant region of the upper airway, and thus more vulnerable to inspiratory collapse, lies cranially to the larynx, the laryngeal pressure-sensing endings seem to be ideally located for detecting collapsing forces and initiating reflex mechanisms for the preservation of patency. This process operates by activating upper airway dilating muscles and by decreasing inspiratory drive: both actions limit t he effect of the collapsing forces. Cold reception is differently represented in various mammalian species within nasal and laryngeal segments. Cooling of the upper airway has an inhibitory influence on breathing, especially in newborns, and a depressive effect on upper airway dilating muscles. The latter response is presumably mediated through the inhibitory effect of cooling on laryngeal pressure endings. These responses could be harmful during occlusive episodes. Powerful defensive responses with distinct characteristics can be elicited through the simulation of laryngeal and nasal irritant type receptors. Sneezing is elicited through the stimulation of trigeminal afferents, cough through the stimulation of laryngeal vagal endings. Changes in osmolality and ionic composition of the mucosal surface liquid can lead to conspicuous alterations in receptor activity and related reflexes.

Laryngeal pressure receptors

We studied the response characteristics of laryngeal pressure receptors in anesthetized dogs, breathing through a tracheal cannula, by recording single unit action potentials from the peripheral cut end of the internal branch of the superior laryngeal nerve. The larynx, with the rest of the upper airway, was isolated and cannulated separately for the application of distending and collapsing pressures. We identified receptors responding to either negative or positive pressure and a few responding to both. All these receptors showed a marked dynamic sensitivity and had the characteristics of slowly adapting mechanoreceptors. The majority of pressure receptors were active at zero transmural pressure and the gain of their response to pressure was higher at lower values, suggesting a role for these receptors in eupnea. Reflex alterations in breathing pattern and upper airway muscle activity during upper airway pressure changes, previously reported, are presumably mediated by the receptors described here. Moreover, these receptors may play a role in certain pathological states, such as obstructive sleep apnea, in which the upper airway is transiently subjected to large collapsing pressure.

Water-responsive laryngeal receptors in the dog are not specialized endings.

The primary purpose of this study was to ascertain whether laryngeal receptors activated by water are specialized endings or whether they also respond to other stimuli, such as pressure, temperature and laryngeal motion as they occur during the breathing cycle. In 35 anesthetized mongrel dogs, breathing spontaneously through a lower cervical tracheostomy, water and other test solutions at approximately 37 degrees C were injected into the functionally isolated larynx with a small catheter. Of the 130 receptors studied, none of the cold receptors (N = 13) responded to water, whereas approximately 60% of all laryngeal mechanoreceptors (72 of 117) responded with either a short delay, short duration or a long delay, long duration response. In general the former pattern of response was exhibited by nonrespiratory-modulated receptors, whereas the latter was typical of respiratory-modulated receptors. The specific nature of the stimulus (hypotonicity or lack of chloride ion) of the water response was further studied in 53 receptors with isoosmotic solutions of dextrose and sodium gluconate. The long delay, long duration response was dependent on a decreased osmolality, while the short delay, short duration response was dependent on the lack of chloride ion of the test solutions. All water-responsive receptors tested (N = 17) were blocked within 50 sec by topically applied 2% lidocaine and thus presumed to be superficial. However, 10 receptors which did not respond to water were also blocked within 50 sec, suggesting that not all superficial receptors are stimulated by water. Based on these observations, we propose that changes in osmolality or ionic composition of the laryngeal surface liquid could play an important role in modifying reflexes involved in the maintenance of upper airway patency.

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.

Circulatory accessibility of nervous receptors localized in the tracheobronchial tree.

We studied the accessibility through the systemic and the pulmonary circulations of slowly and rapidly adapting mechanoreceptors having known location within the tracheobronchial tree of the dog. The accessibility was determined by injecting a challenging substance (veratridine and benzonatate) into the two atria and comparing the delays of the receptor response. Eighteen receptors localized in extrapulmonary airways were more accessible from the systemic circulation; they were affected with a shorter delay following left atrium injection. Six receptors in lobar bronchi and another 44 located in more distal intrapulmonary airways were preferentially accessible from the pulmonary circulation (i.e., shorter delays with right atrium injections). Only 4 receptors localized in lobar bronchi were found to be more accessible from the systemic circulation. Therefore the assumption that airway receptors that are more accessible from the pulmonary circulation are located distal to the terminal bronchioles should be re-evaluated. These results may also have important physiological implications since several endogenous substances capable of affecting airway structures have different concentrations in the mixed venous and arterial blood.

Carbon dioxide-responsive laryngeal receptors in the dog

The purpose of this study was to relate the carbon dioxide (CO2) response of laryngeal receptors to their behavior during the breathing cycle (i.e. their response to transmural pressure changes, laryngeal movement or decreases in temperature) or during exposure to irritant stimuli (water or cigarette smoke). In 9 anesthetized mongrel dogs breathing spontaneously through a tracheostomy, unit activity from the superior laryngeal nerve was recorded while warmed and humidified gas mixtures (air or 10% CO2 in O2) were passed, for 1 min, through the functionally isolated upper airway in the expiratory direction. None of the 10 cold receptors studied were affected by CO2. Eleven of 20 laryngeal non-modulated mechano-receptors were stimulated (from 0.3 to 1.6 imp/sec) by exposure to CO2. These CO2-responsive receptors were also stimulated by known irritant stimuli (cigarette smoke, water), although not all receptors which responded to these irritants were stimulated by CO2. Twelve of 33 respiratory-modulated receptors were affected by CO2; 4 were stimulated and 8 inhibited. Receptors inhibited by CO2 were also inhibited by negative pressure while receptors stimulated by CO2 were also stimulated by negative pressure. These results show that CO2-responsive laryngeal receptors are not specialized endings. Although it is not clear to what extent each separate group of laryngeal receptors is involved, each may contribute to the reflex bradypnea which has been observed during exposure of the upper airway to elevated levels of CO2. However, the importance of CO2-responsive laryngeal receptors in physiological conditions remains unclear.

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.

Menthol in the upper airway depresses ventilation in newborn dogs

Upper airway cooling depresses ventilation in the newborn dog. Since airway cooling stimulates laryngeal cold receptors and inhibits laryngeal mechanoreceptors, the type of afferent ending responsible for this reflex cannot be easily identified. l-menthol, a specific stimulant of cold receptors in the absence of any cooling, has been used to ascertain the discrete role of upper airway cold receptors in this ventilatory depression. Experiments were carried out in 8 anesthetized 7-14-day-old dogs breathing through a tracheostomy with the upper airway functionally isolated. Constant flows of warm air (37 degrees C), with and without addition of l-menthol, and cold air (25 degrees C) were delivered through the upper airway in the expiratory direction. As compared to warm air trials, cold air and warm air + l-menthol trials greatly reduced ventilation (57.5 +/- 10.7% and 52.8 +/- 11.7% of control, respectively; P less than 0.01) mostly due to a prolongation of Te (291.2 +/- 106.4% and 339.2 +/- 90.0%, respectively, P less than 0.01). Section of the superior laryngeal nerve abolished the response to cold air. However, a residual depressive effect of l-menthol was still present in 3 of 5 animals and was abolished by nasal anesthesia, suggesting the involvement of nasal cold receptors. The results suggest that in the newborn dog stimulation of laryngeal cold receptors, without any concurrent inhibition of laryngeal mechanoreceptors, is a sufficient stimulus to cause respiratory depression.

Respiratory Activity of the Cricothyroid Muscle

Single motor units of the cricothyroid muscle (CT), the contralateral CT electromyogram, and the posterior cricoarytenoid muscle (PCA) electromyogram were recorded in anesthetized, spontaneously breathing dogs. In quiet breathing the CT was active predominantly in inspiration, and distinct phasic expiratory activity was observed at lighter levels of anesthesia. Both the CT and PCA increased their inspiratory and expiratory activity with hypercapnia, whereas only their inspiratory activity increased in response to negative pressure and/or absence of volume feedback. Cold blockade of either the recurrent laryngeal nerves or the external branch of the superior laryngeal nerves did not modify CT or PCA activity. In general, activity of CT motor units reflected the behavior of the whole muscle, but different units were recruited at different levels of CT activity. Even though the majority exhibited similar thresholds for inspiration and expiration, some units showed a lower threshold for either one, suggesting some degree of specialization. However, for a few units with high threshold for inspiration the expiratory threshold could not be determined, since a comparable level of CT activity was not achieved in expiration.

Control of postural changes of end expiratory volume (FRC) by airways slowly adapting mechanoreceptors.

We recorded the e.m.g. activity of the diaphragm and of an abdominal muscle (ext. oblique) and the respiratory volume in anesthetized rabbits challenged with head-up tilting and positive pressure breathing (PPB). Both maneuvers determined an inhibition of inspiratory activity and an activation of abdominal muscles, the latter being especially marked with tilting. After cervical vagotomy neither the inspiratory inhibition nor the abdominal recruitment was present during tilting and PPB and the FRC increase was more pronounced. Sulphur dioxide was given in the inspired air (200 ppm) to selectively block the slowly adapting mechanoreceptors. Such blockade was indicated by the absence of the Hering-Breuer inflation reflex. The permanence of other respiratory reflexes was shown by a parodoxical response to inflation and by a still evident response to deflation. With SO2 block, both tilting and PPB did not elicit either the inspiratory inhibition or the abdominal muscles activation, leading to an FRC shift similar to that observed after vagotomy. We conclude that the slowly adapting mechanoreceptors subserve a reflex mechanism relevant in controlling FRC.