Recurrent Laryngeal Nerve Activity Research Articles

Early Changes in Porcine Larynges Following Injection of Motor-Endplate Expressing Muscle Cells for the Treatment of Unilateral Vocal Fold Paralysis.

No curative injectable therapy exists for unilateral vocal fold paralysis. Herein, we explore the early implications of muscle-derived motor-endplate expressing cells (MEEs) for injectable vocal fold medialization after recurrent laryngeal nerve (RLN) injury. Yucatan minipigs underwent right RLN transection (without repair) and muscle biopsies. Autologous muscle progenitor cells were isolated, cultured, differentiated, and induced to form MEEs. Three weeks after the injury, MEEs or saline were injected into the paralyzed right vocal fold. Outcomes including evoked laryngeal electromyography (LEMG), laryngeal adductor pressure, and acoustic vocalization data were analyzed up to 7 weeks post-injury. Harvested porcine larynges were examined for volume, gene expression, and histology. MEE injections were tolerated well, with all pigs demonstrating continued weight gain. Blinded analysis of videolaryngoscopy post-injection revealed infraglottic fullness, and no inflammatory changes. Four weeks after injection, LEMG revealed on average higher right distal RLN activity retention in MEE pigs. MEE-injected pigs on average had vocalization durations, frequencies, and intensities higher than saline pigs. Post-mortem, the MEE-injected larynges revealed statistically greater volume on quantitative 3D ultrasound, and statistically increased expression of neurotrophic factors (BDNF, NGF, NTF3, NTF4, NTN1) on quantitative PCR. Minimally invasive MEE injection appears to establish an early molecular and microenvironmental framework to encourage innate RLN regeneration. Longer follow-up is needed to determine if early findings will translate into functional contraction. NA Laryngoscope, 134:272-282, 2024.

Effect of remimazolam on intraoperative neuromonitoring during thyroid surgery: a case series.

Intraoperative neuromonitoring is widely used to prevent accidental injury during thyroid surgery. Anesthesia should be performed without muscle relaxant or agents with high muscle-relaxant potency. Remimazolam, a novel intravenous anesthetic, became available for clinical use in 2020. Remimazolam is an ultra-short-acting benzodiazepine with a very high clearance rate. However, there are very few data regarding its effect on currently used intraoperative neurological monitoring. Five patients underwent thyroid surgery using intraoperative recurrent laryngeal neuromonitoring. In all cases, intubation was performed after the administration of rocuronium. Anesthesia was maintained by continuous administration of remimazolam at the recommended dose and remifentanil, and no additional rocuronium or sugammadex was administered. Recurrent laryngeal nerve activity could be detected at the first stimulus after surgery was started, and monitoring continued thereafter. Intraoperative monitoring was performed without problems and all surgeries were completed without any complications. Anesthesia with remimazolam at the normal dose did not prolong the time to first positive electromyogram in patients undergoing thyroid surgery, and enables intraoperative recurrent laryngeal nerve monitoring to be performed without any serious perioperative adverse events. Remimazolam may provide a comparable quality of anesthesia to that of existing drugs for neuromonitoring during thyroid surgery.

Laryngospasm, central and obstructive apnea during seizures: Defining pathophysiology for sudden death in a rat model

Seizure spread into the autonomic nervous system can result in life-threatening cardiovascular and respiratory dysfunction. Here we report on a less-studied consequence of such autonomic derangements—the possibility of laryngospasm and upper-airway occlusion.We used parenteral kainic acid to induce recurring seizures in urethane-anesthetized Sprague Dawley rats. EEG recordings and combinations of cardiopulmonary monitoring, including video laryngoscopy, were performed during multi-unit recordings of recurrent laryngeal nerve (RLN) activity or head-out plethysmography with or without endotracheal intubation. Controlled occlusions of a tracheal tube were used to study the kinetics of cardiac and respiratory changes after sudden obstruction.Seizure activity caused significant firing increases in the RLN that were associated with abnormal, high-frequency movements of the vocal folds. Partial airway obstruction from laryngospasm was evident in plethysmograms and was prevented by intubation. Complete glottic closure (confirmed by laryngoscopy) occurred in a subset of non-intubated animals in association with the largest increases in RLN activity, and cessation of airflow was followed in all obstructed animals within tens of seconds by ST-segment elevation, bradycardia, and death. Periods of central apnea occurred in both intubated and non-intubated rats during seizures for periods up to 33s and were associated with modestly increased RLN activity, minimal cardiac derangements, and an open airway on laryngoscopy. In controlled complete airway occlusions, respiratory effort to inspire progressively increased, then ceased, usually in less than 1min. Respiratory arrest was associated with left ventricular dilatation and eventual asystole, an elevation of systemic blood pressure, and complete glottic closure.Severe laryngospasm contributed to the seizure- and hypoxemia-induced conditions that resulted in sudden death in our rat model, and we suggest that this mechanism could contribute to sudden death in epilepsy.

Recurrent laryngeal nerve activity exhibits a 5-HT-mediated long-term facilitation and enhanced response to hypoxia following acute intermittent hypoxia in rat

A progressive and sustained increase in inspiratory-related motor output ("long-term facilitation") and an augmented ventilatory response to hypoxia occur following acute intermittent hypoxia (AIH). To date, acute plasticity in respiratory motor outputs active in the postinspiratory and expiratory phases has not been studied. The recurrent laryngeal nerve (RLN) innervates laryngeal abductor muscles that widen the glottic aperture during inspiration. Other efferent fibers in the RLN innervate adductor muscles that partially narrow the glottic aperture during postinspiration. The aim of this study was to investigate whether or not AIH elicits a serotonin-mediated long-term facilitation of laryngeal abductor muscles, and if recruitment of adductor muscle activity occurs following AIH. Urethane anesthetized, paralyzed, unilaterally vagotomized, and artificially ventilated adult male Sprague-Dawley rats were subjected to 10 exposures of hypoxia (10% O(2) in N(2), 45 s, separated by 5 min, n = 7). At 60 min post-AIH, phrenic nerve activity and inspiratory RLN activity were elevated (39 ± 11 and 23 ± 6% above baseline, respectively). These responses were abolished by pretreatment with the serotonin-receptor antagonist, methysergide (n = 4). No increase occurred in time control animals (n = 7). Animals that did not exhibit postinspiratory RLN activity at baseline did not show recruitment of this activity post-AIH (n = 6). A repeat hypoxia 60 min after AIH produced a significantly greater peak response in both phrenic and RLN activity, accompanied by a prolonged recovery time that was also prevented by pretreatment with methysergide. We conclude that AIH induces neural plasticity in laryngeal motoneurons, via serotonin-mediated mechanisms similar to that observed in phrenic motoneurons: the so-called "Q-pathway". We also provide evidence that the augmented responsiveness to repeat hypoxia following AIH also involves a serotonergic mechanism.

Arterial pulse modulated activity is expressed in respiratory neural output

Although it is well-established that sympathetic activity is modulated with respiration, it is unknown whether neural control of respiration is reciprocally influenced by cardiovascular function. Even though previous studies have suggested the existence of pulse modulation in respiratory neurons, they could not exclude the possibility that such cells were involved in cardiovascular rather than respiratory motor control, owing to neuroanatomic and functional overlaps between brain stem neurons involved in respiratory and cardiovascular control. The aim of this study was to test the hypothesis that respiratory motoneurons and putative premotoneurons are modulated by arterial pulse. An existing data set composed of 72 well-characterized, respiratory-modulated brain stem motoneurons and putative premotoneurons was analyzed using delta(2), a recently described statistic that quantifies the magnitude of arterial pulse-modulated spike activity [Dick TE and Morris KF. J Physiol 556: 959-970, 2004]. Neuronal activity was recorded in the rostral and caudal ventral respiratory groups of 19 decerebrate, neuromuscular-blocked, ventilated cats. Axonal projections were identified by rectified and unrectified spike-triggered averages of recurrent laryngeal nerve activity or by antidromic activation from spinal stimulation electrodes. The firing rates of approximately 30% of these neurons were modulated in phase with both the respiratory and cardiac cycles. Furthermore, arterial pulse modulation occurred preferentially in the expiratory phase in that only expiratory neurons had high delta(2) values and only expiratory activity had significant delta(2) values after partitioning tonic activity into the inspiratory and expiratory phases. The results demonstrate that both respiratory motoneurons and putative premotoneuronal activity can be pulse modulated. We conclude that a cardiac cycle-related modulation is expressed in respiratory motor activity, complementing the long-recognized respiratory modulation of sympathetic nerve activity.

Glycinergic inhibition is essential for co-ordinating cranial and spinal respiratory motor outputs in the neonatal rat.

Eupnoeic breathing in mammals is dependent on the co-ordinated activity of cranial and spinal motor outputs to both ventilate the lungs and adjust respiratory airflow, which they do by regulating upper-airway resistance. We investigated the role of central glycinergic inhibition in the co-ordination of cranial and spinal respiratory motor outflows. We developed an arterially perfused neonatal rat preparation (postnatal age 0-4 days) to assess the effects of blocking glycine receptors with systemically administered strychnine (0.5-1 microM). We recorded respiratory neurones located within the ventrolateral medulla, inspiratory phrenic nerve activity (PNA) and recurrent laryngeal nerve activity (RLNA), as well as dynamic changes in laryngeal resistance. Central recordings of postinspiratory neurones revealed an earlier onset in firing relative to the onset of inspiratory PNA after exposure to strychnine (260 +/- 38.9 vs. 129 +/- 26.8 ms). After glycine receptor blockade, postinspiratory neurones discharged during the inspiratory phase. Strychnine also evoked a decrease in PNA frequency (from 38.6 +/- 4.7 to 30.7 +/- 2.8 bursts min(-1)), but amplitude was unaffected. In control conditions, RLNA comprised inspiratory and postinspiratory discharges; the amplitude of the latter exceeded that of the former. However, after administration of strychnine, the amplitude of inspiratory-related discharge increased (+65.2 +/- 15.2 %) and exceeded postinspiratory activity. Functionally this change in RLNA caused a paradoxical, inspiratory-related glottal constriction during PNA. We conclude that during the first days of life in the rat, glycine receptors are essential for the formation of the eupnoeic-like breathing pattern as defined by the co-ordinated activity of cranial and spinal motor inspiratory and postinspiratory activities.

Medullary respiratory neurones and control of laryngeal motoneurones during fictive eupnoea and cough in the cat.

1. This study addressed the hypothesis that ventrolateral medullary respiratory neurones participate in the control of laryngeal motoneurones during both eupnoea and coughing. 2. Data were obtained from 28 mid-collicular decerebrated, artificially ventilated cats. Cough-like motor patterns (fictive cough) in phrenic, lumbar and recurrent laryngeal nerves were elicited by mechanical stimulation of the intrathoracic trachea. Microelectrode arrays were used to monitor simultaneously several neurones in the ventral respiratory group, including the Bötzinger and pre-Bötzinger complexes. Spike trains were evaluated for responses during fictive cough and evidence of functional connectivity with spike-triggered averages of efferent recurrent laryngeal nerve activity. 3. Primary features were observed in averages triggered by 94 of 332 (28 %) neurones. An offset biphasic wave with a positive time lag was present in the unrectified average for 10 inspiratory and 13 expiratory neurones. These trigger neurones were respectively identified as inspiratory laryngeal motoneurones with augmenting, decrementing, plateau and "other" discharge patterns, and expiratory laryngeal motoneurones with decrementing firing patterns. 4. Rectified averages triggered by inspiratory neurones included 37 offset peaks, 11 central peaks and one offset trough. Averages triggered by expiratory neurones had 12 offset peaks, six central peaks and four offset troughs. Relationships inferred from these features included premotor actions of inspiratory neurones with augmenting, decrementing, plateau and "other" patterns on inspiratory laryngeal motoneurones, and premotor actions of decrementing and "other" expiratory neurones on expiratory laryngeal motoneurones. Corresponding changes in neuronal firing patterns during fictive cough supported these inferences. 5. The data confirm and extend previous results on the control of laryngeal motoneurones during eupnoea and support the hypothesis that the same premotor neurones help to shape motoneurone firing patterns during both eupnoea and coughing.

Central mechanisms controlling upper airway patency

The neurones of the ponto-medullary respiratory network drive two functionally and anatomically distinct pools of motoneurones. One set is located within the spinal cord that innervates the diaphragm and intercostal muscles. A second group of motoneurones is located within the nucleus ambiguus imbedded in the ventral respiratory group that project via cranial motor outflows to co-ordinate the activity of laryngeal and bronchial muscles to control airway resistance and airflow. These spinal and cranial motor activities have to be precisely co-ordinated to ensure efficient ventilation. We recently included this behaviour as imperative for a definition of eupnoea. Thus, beside a rhythmic eupnoeic (ramp) discharge pattern of pump motoneurones, a phasic respiratory modulation of glottal resistance should be observed and expressed as glottal dilation during inspiration and transient glottal constriction during post-inspiration [1]. During the last decade mechanisms underlying respiratory rhythm generation were studied primarily in reduced in vitro preparations. The work concluded that the respiratory rhythm is generated by pacemaker neurones located in the Pre-Botzinger complex and is independent of inhibitory glycinergic synaptic transmission (for review see [2,3]). However, a potential role for glycinergic transmission for the eupneic co-ordination of circuitry controlling the upper airway was largely disregarded. To determine a role for glycinergic inhibition within the pon-tomedullary network, we used the arterially perfused working heart-brainstem preparation (WHBP) of neonatal and mature rat. This preparation allows both kinesiological and cellular studies of central and peripheral mechanisms controlling upper airway resistance [1]. Recording of the recurrent laryngeal nerve activity as an index of motor output to the glottis revealed post-inspiratory activity that shifted towards the inspiratory phase after strychnine antagonism of glycine receptors (0.5–1.5 μM). This shift of post-inspiratory activity was also obtained at the cellular level: intracellular recordings of post-inspiratory neurones revealed that the hyperpo-larisation during the inspiratory phase was converted to a depolarisation with spike discharge after exposure to strychnine. This lead to a massive disturbance of the eupnoeic modulation of glottal resistance by converting the inspiratory glottic dilatation (seen during control) to a paradoxical constriction, as demonstrated by measuring changes in laryngeal resistance. Similar results were obtained in both neonatal and juvenile rats suggesting that glycinergic mechanisms co-ordinating ventilatory movements with upper airway resistance are functional at birth. The effects of glycinergic inhibition were mimicked during exposure of neonatal preparations to prolonged hypoxia. During the secondary hypoxic depression of respiration post-inspiratory activity was shifted towards inspiration causing a paradoxical glottic constriction during neural inspiration. We conclude that integrity of glycinergic neurotransmission within the ponto-medullary respiratory network is essential for co-ordinating the neuronal activities which control upper airway resistance and ventilatory movements and consequently the eupnoeic breathing pattern in rats from birth.

Functional connectivity among ventrolateral medullary respiratory neurones and responses during fictive cough in the cat.

This study tested predictions from a network model of ventrolateral medullary respiratory neurone interactions for the generation of the cough motor pattern observed in inspiratory and expiratory pump muscles. Data were from 34 mid-collicularly decerebrated, paralysed, artificially ventilated cats. Cough-like patterns (fictive cough) in efferent phrenic and lumbar nerve activities were elicited by mechanical stimulation of the intrathoracic trachea. Neurones in the ventral respiratory group, including the Botzinger and pre-Botzinger complexes, were monitored simultaneously with microelectrode arrays. Spike trains were analysed for evidence of functional connectivity and responses during fictive cough with cycle-triggered histograms, autocorrelograms, cross-correlograms, and spike-triggered averages of phrenic and recurrent laryngeal nerve activities. Significant cross-correlogram features were detected in 151 of 1988 pairs of respiratory modulated neurones. There were 59 central peaks, 5 central troughs, 11 offset peaks and 2 offset troughs among inspiratory neurone pairs. Among expiratory neurones there were 23 central peaks, 8 offset peaks and 4 offset troughs. Correlations between inspiratory and expiratory neurones included 20 central peaks, 10 central troughs and 9 offset troughs. Spike-triggered averages of phrenic motoneurone activity had 51 offset peaks and 5 offset troughs. The concurrent responses and multiple short time scale correlations support parallel and serial network interactions proposed in our model for the generation of the cough motor pattern in the respiratory pump muscles. Inferred associations included the following. (a) Excitation of augmenting inspiratory (I-Aug) neurones and phrenic motoneurones by I-Aug neurones. (b) Inhibition of augmenting expiratory (E-Aug) neurones by decrementing inspiratory (I-Dec) neurones. (c) Inhibition of I-Aug, I-Dec and E-Aug neurones by E-Dec neurones. (d) Inhibition of I-Aug and I-Dec neurones and phrenic motoneurones by E-Aug neurones. The data also confirm previous results and support hypotheses in current network models for the generation of the eupnoeic pattern.

Effects of driving pressure and recurrent laryngeal nerve stimulation on glottic vibration in an in vivo constant pressure model of phonation.

Most in vivo studies of phonation have used constant flow models. However, because the lung–thorax system is better viewed as a constant pressure source, a constant pressure in vivo canine model was developed. In this model, driving pressure and recurrent laryngeal nerve (RLN) stimulation are independent variables, and subglottic pressure (Psub; measured immediately under the glottis), air flow, fundamental frequency (F0), and glottic area are dependent variables. In three dogs, Psub and air flow were measured with constant RLN stimulation and varying driving pressure, and in another condition with constant driving pressure and varying RLN stimulation. Videostroboscopic measures on four animals assessed glottic areas with constant RLN stimulation and varying driving pressure. With constant RLN stimulation, increasing driving pressure had no effect on glottic area, while F0, and air flow increased significantly. Changes in Psub were small in comparison to changes in driving pressure. With constant driving pressure, increasing RLN stimulation increased Psub and F0 and decreased air flow. These findings suggest that Psub during phonation is primarily dependent on RLN activity and associated laryngeal muscular contraction, but not on lung driving pressure. [Work supported by VA Merit Review funds.]

Changes in phrenic, hypoglossal and recurrent laryngeal nerve activities after intravenous infusions of aminophylline in cats

Aminophylline is known to have respiratory stimulant properties, and it has been suggested that it may also be effective in sleep apnoea. However, its role in this disorder remains uncertain. Theoretically, increasing upper airway motoneural activity in order to maintain airway patency might alleviate obstructive sleep apnoea. On the other hand, increasing the respiratory drive may also prove beneficial in treating central sleep apnoea. In these studies, we attempted to determine the effect of aminophylline on neural activities of the upper airway and diaphragm. We administered intravenously either a low dose (4 mg.kg-1) or a high dose (16 mg.kg-1) of aminophylline to decerebrated, vagotomized and paralysed cats, and continuously recorded the phrenic hypoglossal and recurrent laryngeal nerve activities for 3 h. Results showed that a high dose of aminophylline induced a marked increase in phrenic nerve activity, but not hypoglossal or recurrent laryngeal nerve activity. In a group treated with a low dosage of aminophylline, a significant increase of activity was found in all three nerves. Furthermore, phrenic nerve activity increased more with a high dose than with a low dose. We confirmed that aminophylline has dose-dependent and selective effects on respiratory neural activity. A low dose acts on the upper airway and diaphragm, but a high dose induces a marked increase in central respiratory drive. According to our results, low dose aminophylline might be beneficial in obstructive sleep apnoea, whereas, a high or low dose might improve some cases of central sleep apnoea.

Phrenic and recurrent laryngeal motoneuron activities during hyperoxia and hypoxia in piglets

We hypothesized that synchronization of inspiratory motoneurons may involve inputs from two central pattern generators (CPG): one characterized by medium-frequency (< 50 Hz) and the other by high-frequency oscillations (≧ 50 Hz). We studied phrenic and recurrent laryngeal nerve activities recorded during hyperoxia and hypoxia in Saffan anesthetized, paralyzed, and artificially ventilated piglets. Spectral analyses, derived from the full as well as partitioned halves of inspiration, showed that phrenic and recurrent laryngeal discharges contained peaks in the medium-frequency band, which were indicative of common inputs. The phrenic spectra of many animals had peaks in the high-frequency band; such peaks were uncommon in recurrent laryngeal spectra; consequently, correlated activities corresponding to high-frequency oscillations were not usually observed. Thus, it is likely that acquisition of modulating inputs from a high-frequency CPG may emerge in an age-dependent manner in different motoneuron pools. During hypoxia, both phrenic and recurrent laryngeal discharges were facilitated as shown by increases in both the amplitudes of signal-averaged histograms and the magnitudes of their respective power spectral activities. Also, there was a significant increase in the values of phrenic-recurrent laryngeal coherence estimates in the medium-frequency region. Hence, medium-frequency oscillations are more apparent in early development, perhaps to facilitate synchronization of inspiratory motoneuron activities, especially under conditions of increased chemical drive.

Synaptic interaction between medullary respiratory neurones during apneusis induced by NMDA-receptor blockade in cat.

1. Termination of inspiration is an essential component of respiratory rhythm generation and its perturbation can result in apneusis, i.e. significant prolongation of mechanisms, we studied the postsynaptic events in respiratory neurones during apneustic respiratory periods, and compared them to normal respiratory cycles. 2. Experiments were performed in pentobarbitone-anaesthetized, paralysed, thoracotomized cats ventilated with a constant volume or a cycle-triggered constant pressure pump. Apneusis, separated by normal cycles, was induced as follows: the animal was ventilated by a cycle-triggered pump that normally inflated the lungs during the inspiratory burst of phrenic nerve discharge. The NMDA-receptor blocker MK-801 [(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5, 10-iminemaleate] (0.3-0.7 mg/kg) was administered intravenously, and, for designated breaths, inflation of the lungs was withheld during neural inspiration. 3. Membrane potential trajectories of forty-one late expiratory (E-2) and eight postinspiratory (PI) neurones of the caudal ventral respiratory group were analysed before and/or after MK-801 administration, during normal and apneustic periods. 4. Before MK-801 administration, withholding lung inflation caused modest (10-20%) lengthening of the inspiratory period; after MK-801 administration, withholding inflation caused apneusis. Provided that the lungs were inflated during the inspiratory phase, the temporal pattern of phrenic nerve, recurrent laryngeal nerve and membrane potential trajectories of E-2 and PI neurones were not significantly altered by MK-801. Apneusis following NMDA-receptor blockade produced consistent changes in the synaptic activation patterns of E-2 neurones. In particular, the slow late inspiratory-related depolarization pattern of E-2 neurones was consistently retarded during apneustic inspiratory phases when compared to normal inspiratory phases. This was due to continuation of Cl(-)-mediated synaptic inhibition of E-2 neurones. Superior laryngeal nerve stimulation stopped apneusis and sustained membrane hyperpolarization of E-2 neurones similar to lung inflation. 5. During the plateau phase of apneusis, correlated 10-20 Hz oscillations could be observed in the integrated phrenic and recurrent laryngeal nerve activities as well as in the membrane potential of E-2 neurones. 6. We conclude that: (i) the prolonged inhibition of E-2 neurones during apneusis is indicative of the process responsible for the prolongation of the inspiratory phase.(ABSTRACT TRUNCATED AT 400 WORDS)

Responses to severe hypoxia of phrenic and recurrent laryngeal nerve activity in vagotomized cats.

We investigated changes in activities of phrenic nerve (PN) and the recurrent laryngeal nerve (RLN) during progressive hypoxia produced by administration of a mixture of 5% O2 in N2 and a mixture of 5% O2 in N2O in 8 vagotomized, paralyzed, and artificially ventilated cats anesthetized with halothane. During progressive hypoxia produced by administration of 5% O2 in N2, both PN and RLN activities initially increased and then decreased at approximately the same rate. The relationship between PN and RLN activities during the respiratory stimulation and the relationship between PN and RLN activities during the depression due to hypoxia were both linear and were represented by the same linear regression line. The responses of PN and RLN activities to progressive hypoxia produced by administration of 5% O2 in N2O were basically similar to those observed during administration of 5% O2 in N2 although a concomitant increase in depth of anesthesia with N2O enhanced the occurrence of hypoxic respiratory depression. These results suggest that the respiratory modulation of recurrent laryngeal motoneuron activity is closely related to that of phrenic motoneuron activity and that both motoneurons share similar control mechanisms. Neither severe hypoxia nor addition of N2O to a halothane-anesthetized cat seems to affect the close linear relationship between PN and RLN activities.

Interaction of hypercapnia and phasic volume feedback on motor control of the upper airway.

The effect of hypercapnia on the suppression of efferent hypoglossal and recurrent laryngeal nerve activity by phasic volume feedback was studied in decerebrate paralyzed intubated cats ventilated with a phrenic-driven servo-respirator. The gain of the respirator was altered for single inspirations, and the resulting changes in neural activities were quantified by comparison with respective neural activities without phasic volume feedback. This maneuver was performed when the end-tidal CO2 concentration was 5, 7, and 9%. Changes in the level of CO2 did not alter the slope or position of the volume thresholds for suppression of hypoglossal and recurrent laryngeal activities. The slope of the volume-time isopleths for specific levels of graded suppression also remained constant for each nerve at the different levels of CO2. Under hypercapnic conditions, greater volumes were required at a given time into inspiration to achieve any particular level of suppression, but these differences generally did not reach statistical significance. These data demonstrate a lack of effect of the CO2 stimulus on the suppression of upper airway motoneuron activity by phasic volume feedback. Despite the absence of this interaction, a CO2-induced increase in central inspiratory activation of upper airway motoneurons, in the presence of a very sensitive volume feedback system, would help maintain airway patency in the face of upper airway narrowing or closure.

Split medulla preparation in the cat: arterial chemoreceptor reflex and respiratory modulation of the renal sympathetic nerve activity

The study was undertaken in order to assess the changes in sympathetic output in a split medulla preparation of the cat which, as shown earlier, has impaired respiratory rhythm generation. The effects of medullary midsagittal sections on renal sympathetic nerve firing were investigated in chloralose anesthetized, paralyzed and artificially ventilated cats. Recordings of phrenic and recurrent laryngeal nerve activity served as indices of central respiratory rhythm generation. Sections, 5 mm deep from the dorsal medullary surface and extending 6 mm rostrally and 3 mm caudally to the obex, did not produce any significant changes in heart rate, blood pressure or tonic renal sympathetic nerve firing levels. They decreased or abolished, however, the respiratory rhythmicity in renal sympathetic nerve which paralleled the reduction of inspiratory discharges in phrenic and recurrent laryngeal nerves, and abolished the carotid body chemoreceptor-sympathetic reflex. The inspiratory activity remaining after the sections could still be enhanced by chemoreceptor stimulation. The inhibitory baroreceptor and pulmonary stretch receptor sympathetic reflexes, and the central excitatory effect of CO 2 on renal sympathetic nerve firing were not altered. The effects of electrical stimulation within the midsagittal plane of the medulla have shown that descending pathways from the medullary inspiratory neurons (or their medullary collaterals) do not participate in the facilitation of spinal preganglionic neurons during inspiration and in relaying the pulmonary stretch receptor inhibitory sympathetic reflex. A region located close to the obex was identified from which excitatory responses in renal sympathetic nerves, compatible with the response obtained by carotid sinus nerve stimulation, could be evoked. It is concluded that a lesion in the midline of the lower medulla at the level of the obex selectively destroys cells or pathways which relay the carotid body chemoreceptor-sympathetic reflex.

Differential elevation by protriptyline and depression by diazepam of upper airway respiratory motor activity.

Effects of systemically administered protriptyline and diazepam on the respiratory activity of the phrenic, hypoglossal, and recurrent laryngeal nerves were investigated in vagotomized, decerebrate cats. Both hypoglossal and recurrent laryngeal nerve activities were consistently increased after protriptyline administration, whereas the phrenic nerve discharge was not systematically altered. Similar changes were observed in cats with bilateral carotid sinus nerve sections. Diazepam induced a reduction of hypoglossal and recurrent laryngeal nerve activities at doses that did not alter phrenic nerve discharge. These results with diazepam were the same in carotid chemodenervated cats. We conclude that neural mechanisms controlling upper airway muscles are much more sensitive to protriptyline and diazepam than are those of the bulbospinal-phrenic system. The selective augmentation of hypoglossal and recurrent laryngeal discharges by protriptyline could account for the reported decrease in the frequency of obstructive sleep apneas in patients receiving this antidepressant. In contrast, diazepam, by depressing motor activity to upper airway muscles, may exacerbate oropharyngeal obstruction during sleep.

Selective depression by ethanol of upper airway respiratory motor activity in cats.

We studied the effects of systemically administered ethyl alcohol on the respiratory motor activity of the phrenic, hypoglossal and recurrent laryngeal nerves in unanesthetized, decerebrate cats. Some of the cats were studied after carotid sinus nerve section. In addition, parallel studies were done in intact, awake cats with chronic electromyographic electrodes in the diaphragm, genioglossus, and posterior cricoarytenoid (PCA) muscles. In decerebrate animals, alcohol induced a significant reduction of hypoglossal and recurrent laryngeal nerve activities at doses that had little or no effect on the phrenic nerve discharge. Similar changes were observed in chemodenervated cats. In awake animals, genioglossal and PCA muscle activities were depressed by alcohol, whereas diaphragm activity showed no consistent change. Alcohol caused a significant increase in respiratory frequency in awake cats and reduced the responses of genioglossal and PCA muscle activities to hypercapnia and normocapnic hypoxia. We conclude that alcohol induces a selective reduction in upper airway respiratory motor activity by an action that does not require intact suprapontile structures, vagal afferents, or peripheral chemoreceptors. This reduction may contribute to the alcohol-induced exacerbation of obstructive sleep apnea.

Power spectral analysis of inspiratory nerve activity in the decerebrate cat

To investigate the high frequency oscillations observed in the inspiratory activity of respiratory motor nerves of decerebrate cats, we applied a signal processing technique, power spectral analysis, to the electrical activity of the phrenic and recurrent laryngeal nerves. We found two peaks in the phrenic nerve power spectral densities, one at 88.1 ± 6.4 Hz (mean ± S.D.) and the other at 37.1 ± 9.7 Hz, and two peaks for the recurrent laryngeal nerve, at 87.4 ± 10.1 Hz and at 55.4 ± 5.1 Hz. We identified 3 factors affecting the peaks. Anesthetics reduced or eliminated the 88 Hz peak and produced new low frequency peaks in the phrenic and recurrent laryngeal nerves. Increasing end-tidal CO 2 decreased the bandwidth of the 88 Hz peak and increased its amplitude relative to that of the low frequency peak. Decreasing body temperature from 38 to 30°C reduced the frequency of the 88 Hz peak by 5.0 Hz/°C. The power spectral density of the phrenic nerve activity differed from that of the recurrent laryngeal nerve activity because the single fibers in each nerve had different power spectral densities. About 70% of the fibers recorded in a nerve had power spectral densities similar to that of the whole nerve. A minority of the phrenic nerve fibers had the same low spectral peak as the recurrent laryngeal fibers had the same low spectral peak as the a minority of the recurrent laryngeal fibers had the same low spectral peak as the phrenic nerve. Bilateral removal of the dorsal respiratory group eliminated the high frequency peak in the power spectral density of the phrenic nerve and the peripheral reflexes, but rhythmic bursts of inspiratory activity remained. From these findings we hypothesized that there are two central respiratory pattern generators in the brain stem with parallel pathways to the respiratory motoneurons.

Aortic nerve stimulation in the rat: Cardiovascular and respiratory responses

Cardiovascular response and changes in the phrenic and recurrent laryngeal nerve activities to the electrical stimulation of the aortic nerves were studied in the decerebrate male Wistar rats. Stimulus parameters were varied over a wide range (intensities 0.1-10 volts, frequencies 2-150 pulses per sec, durations 0.01-10 msec) and stimulation was continued for 1 min to ensure that steady state was reached. Stimulation of the aortic nerves consistently produced decreases in blood pressure and heart rate (baroceptor responses) while increase in blood pressure and heart rate (chemoreceptor responses) were never obtained. Inhibition of phrenic and recurrent laryngeal nerve activities (baroceptor responses) always followed stimulation of aortic nerves while facilitation of these nerve activities (chemoreceptor responses) was never observed. These results indicate that the aortic nerves of the rat contain mainly baroceptor afferent fibers and there are few, if any, functional chemoreceptor fibers in these nerves.