Analysis of the respiratory role of intrinsic laryngeal motoneurons of cat.

The electromyographic activity of the posterior cricoarytenoid, thyroarytenoid, lateral cricoarytenoid muscles, and the phrenic electroneuronogram were recorded along with measurements of tidal inspiration and trachael airflow in chloralose-urethane anesthetized cats. In eupnea, the posterior cricoarytenoid exhibited tonic and inspiratory phasic activity; the other two muscles showed only expiratory bursts of activity. Lung inflation inhibited phasic posterior cricoarytenoid activity to reveal a tonic component; activity of the other two was inhibited. Lung deflation increased activity in all three. Vagotomy abolished static volume effects and increased spontaneous cyclic activity of laryngeal muscles. Hypercapnia increased posterior cricoarytenoid activity, but decreased thyroarytenoid activity; these effects were enhanced after vagotomy. Hering-Breuer reflexes curb intrinsic laryngeal motoneuronal excitability. The hypercapnic effect on laryngeal motoneurons promotes the egress of CO₂ by widening the glottis due to both facilitation of abduction and attenuation of adduction.     

Reinnervation of laryngeal muscles: a study of changes in myosin heavy chain expression

Direct repair of the recurrent laryngeal nerve (RLN) results in synkinesis and compromised laryngeal function. We have therefore developed a pig model to investigate whether anastomosis of the phrenic nerve with the abductor branch of the RLN leads to specific reinnervation of abductor muscles. Expression of myosin heavy chain protein (MyHC), a marker of appropriate reinnervation, was determined in the posterior cricoarytenoid (PCA) abductor and thyroarytenoid (TA) adductor muscles following nerve injury and repair. The denervated PCA muscle exhibited decreased levels of the fast-type MyHC isoforms IIA and IIB, and increased slow-type MyHC expression. Similarly, there was a fall in type IIB levels in the denervated TA muscle but increases in both IIA and slow MyHC. Four months after repair, the MyHC expression in the PCA was near normal, suggesting that our model reduces the risk of synkinesis and ensures the accurate muscle reinnervation required for full functional recovery.     

Effect of neurotrophin-3 on reinnervation of the larynx using the phrenic nerve transfer technique

Current techniques for reinnervation of the larynx following recurrent laryngeal nerve (RLN) injury are limited by synkinesis, which prevents functional recovery. Treatment with neurotrophins (NT) may enhance nerve regeneration and encourage more accurate reinnervation. This study presents the results of using the phrenic nerve transfer method, combined with NT-3 treatment, to selectively reinnervate the posterior cricoarytenoid (PCA) abductor muscle in a pig nerve injury model. RLN transection altered the phenotype and morphology of laryngeal muscles. In both the PCA and thyroarytenoid (TA) adductor muscle, fast type myosin heavy chain (MyHC) protein was decreased while slow type MyHC was increased. These changes were accompanied with a significant reduction in muscle fibre diameter. Following nerve repair there was a progressive normalization of MyHC phenotype and increased muscle fibre diameter in the PCA but not the TA muscle. This correlated with enhanced abductor function indicating the phrenic nerve accurately reinnervated the PCA muscle. Treatment with NT-3 significantly enhanced phrenic nerve regeneration but led to only a small increase in the number of reinnervated PCA muscle fibres and minimal effect on abductor muscle phenotype and morphology. Therefore, work exploring other growth factors, either alone or in combination with NT-3, is required.     

Crossed and uncrossed central effects of muscle spindle afferents from the lateral pterygoid muscle of the guinea pig

Physiological evidence is presented for the presence of stretch reflexes in the lateral pterygoid (Pt) muscle of the guinea pig. The central reflex effects of excitation of Pt stretch reflex afferents were also investigated. Passive lateral jaw displacement, which resulted in stretch of the Pt muscle on the side of jaw movement and stretch of the zygomatico-mandibularis (Z) muscle on the side contralateral to the movement, evoked increased EMG activity in these muscles. Stimulation of the trigeminal mesencephalic nucleus (mes V) evoked monosynaptic reflexes in both the Pt and Z nerves. Tonic stretch of the Pt muscle facilitated the monosynaptic reflex in the Pt nerve evoked by stimulation of mes V. Tonic vibration of the Pt muscle facilitated the mes V evoked monosynaptic reflex in the nerves to the ipsilateral Pt and contralateral Z muscles. Conversely, tonic vibration of the Z muscle facilitated the monosynaptic reflex evolved by mes V stimulation in the contralateral Pt and ipsilateral Z nerve. The results support the view that muscle spindles exist in the Pt and Z muscles and that there is a monosynaptic stretch reflex for both the Pt and Z muscles with cell bodies located in the mes V nucleus. It was also shown that the ipsilateral Pt muscle and the contralateral Z muscle act as synergists in the production of lateral jaw movements and that the organization of the stretch reflexes originating from the Pt and Z muscles support their synergistic action.     

Analysis of the respiratory role of pharyngeal constrictor motoneurons of cat

The electromyogram of the middle pharyngeal constrictor muscle and the electroneurogram of the phrenic nerve was recorded in parallel with measurements of tidal inspiration and tracheal airflow in chloralose-urethane anesthetized cats. In eupnea, pharyngeal constrictor exhibited tonic and expiratory phasic activity. Lung inflation inhibited phasic pharyngeal constrictor activity to reveal its tonic component; lung deflation eliminated both phasic and tonic components. Vagotomy eliminated these static lung volume effects and increased spontaneous cyclic activity. Hypocapnia abolished phasic activity and unmasked its tonic component in vagotomized cats. Hypercapnia increased cyclic pharyngeal constrictor activity (vagi intact or cut). Weak superior laryngeal or glossopharyngeal nerve stimulation had little or no effect during inspiration; during expiration, superior laryngeal nerve stimulation evoked a short lasting attenuating effect, and glossopharyngeal nerve stimulation exerted a long lasting blocking effect. (vagi intact or cut). Pharyngeal constrictor, like laryngeal adductors, is classified as an expiratory resisting type muscle. Pharyngeal constrictor reduces dead space during hypercapnia thereby promoting the exodus of CO₂. Comparison of vagal and other respitatory motoneurons reveals some special features of neural control of respiration heretofore overlooked.     

Nucleus ambiguus motoneurons innervating the canine intrinsic laryngeal muscles by the fluorescent labeling technique

The localization of motoneurons innervating the canine intrinsic laryngeal muscles was investigated by the fluorescent labeling technique. Labeled cells were found in the ipsilateral nucleus ambiguus. The most rostral labeled neurons for the cricothyroid muscle, the posterior cricoarytenoid muscle, the thyroarytenoid muscle, and the lateral cricoarytenoid muscle were found at progressively more caudal levels, respectively, within the nucleus ambiguus. The rostral tip of the arytenoid muscle cell column was at the same level as the lateral cricoarytenoid muscle cell column. The cells labeled from the cricothyroid muscle occupied the ventral part of the nucleus at the rostral level of the nucleus. At the middle level of the nucleus, the cells from the posterior cricoarytenoid muscle occupied the ventral part of the nucleus and the cells from the thyroarytenoid muscle, the lateral cricoarytenoid muscle and the arytenoid muscle occupied the dorsal part of the nucleus. The existence of double-labeled cells which innervated both thyroarytenoid muscle and lateral cricoarytenoid muscle was detected.     

Localization of rabbit laryngeal motoneurons in the nucleus ambiguus

The localization of rabbit laryngeal motoneurons in nucleus ambiguus was investigated using injection of a fluorescent labeling substance, i.e., nuclear yellow, into the individual laryngeal muscles. The nucleus ambiguus of the rabbit comprises four subnuclei, CoG, SGm, SGl, and DiG. The CoG is a group of compactly arranged neurons, and is situated in the rostral one-half of the nucleus. The SG, situated in its rostral one-third, is scattered around the CoG, with a subdivision into SGm and SGl. These subdivisions are medial and lateral to the CoG, respectively. The DiG is formed by diffusely arranged neurons, and is located in the caudal two-thirds of the nucleus. All labeled motoneurons were found in the ipsilateral nucleus ambiguus. The motoneurons supplying the cricothyroid muscle, which is innervated by the superior laryngeal nerve, were present in the SGm, with a clear rostralward segregation from the other motoneurons. The motoneurons supplying the muscles innervated by the inferior laryngeal nerve were located in the DiG, where they displayed a rostrocaudal myotopical arrangement in the order posterior cricoarytenoid, thyroarytenoid, and lateral cricoarytenoid. The posterior cricoarytenoid motoneurons were intermingled with the thyroarytenoid motoneurons in the rostral two-thirds of the DiG, and the former tended to be concentrated more rostrally than the latter. The lateral cricoarytenoid motoneurons were confined to the most caudal one-third of the DiG.     

Fibers supplying the laryngeal musculature in the cranial root of the rabbit accessory nerve: Nucleus of origin, peripheral course, and innervated muscles

The location of the rabbit laryngeal motoneurons whose axons traverse the cranial root of the accessory nerve was studied with injection of HRP or nuclear yellow into the laryngeal muscles in combination with the intracranial severing of either the rootlets of the vagus nerve or those of the cranial root. The motoneurons were located in the diffuse cell group that forms a subnucleus occupying the caudal two-thirds of the nucleus ambiguus and sending fibers to the inferior laryngeal nerve. The caudal one-third of the diffuse cell group supplying the lateral cricoarytenoid muscle, was occupied only by these motoneurons, whereas in its rostral two-thirds, they were intermingled with motoneurons having axons that traversed the vagal rootlets. The thyroarytenoid and posterior cricoarytenoid motoneurons are present in the rostral two-thirds of the diffuse cell group; axons of the former traversed the rootlets of the cranial root, and of the latter traversed the vagal rootlets. On the other hand, the medial scattered cell group, located in the rostral one-third of the nucleus ambiguus and sending fibers to the cricothyroid muscle via the superior laryngeal nerve, contained only motoneurons with axons traversing the vagal rootlets. The above findings clarified that fibers of the cranial root enter the inferior laryngeal nerve after joining the vagus, and then reach the adductor muscles for the vocal fold, with their neurons of origin in a caudal portion of the nucleus ambiguus. The vagal rootlet fibers, with their neurons of origin situated more rostrally in the nucleus, innervate the tensor and abductor muscles via the superior and inferior laryngeal nerve, respectively.     

An implantable system for In Vivo chronic electromyographic study in the larynx

Introduction: Electromyography (EMG) plays an important role in exploring the mechanisms of selective reinnervation. An implantable system can help provide chronological information regarding reinnervation of laryngeal muscles. This study was designed to develop an implantable system for repeated recordings of spontaneous and evoked EMG from laryngeal muscles. Methods: This implantable system has 4 bipolar stimulus cuffs for bilateral recurrent laryngeal nerves (RLNs) and superior laryngeal nerves (SLNs), and 4 EMG recording electrodes for bilateral vocal fold adductors (thyroarytenoid–lateral cricoarytenoid, TA‐LCA) and abductor (posterior cricoarytenoid, PCA) muscles. The system was implanted in 8 canines for up to 41 weeks. Results: The system showed good compatibility. Consistent EMG signals were recorded from both PCA and TA‐LCA muscles. Conclusion: We developed a long‐term implantable EMG system that is simple and capable of obtaining stable EMG recordings from canine laryngeal muscles with minimal risk of device breakage, trauma, or infection. Muscle Nerve 55: 706–714, 2017     

Differential inhibition of the diaphragm and posterior cricoarytenoid muscles induced by transient hypertension across sleep states in intact cats

Arterial pressure was transiently elevated by intravenous infusion of phenylephrine in intact, freely moving cats during sleep and waking states to determine pressure effects on diaphragmatic and laryngeal abductor EMG activity. Transient hypertension caused respiratory cycle duration to increase and integrated EMG area to decrease for several breaths in both the diaphragm and posterior cricoarytenoid, the integrated inspiratory area of which decreased to a greater extent than did that of the diaphragm. Cycle duration increases resulted from increases in expiratory duration. Expiratory duration of the posterior cricoarytenoid initially increased proportionately more than that of the diaphragm, causing a transient phase disassociation between that upper airway muscle and diaphragmatic timing. This disassociation disappeared after several breaths. Changes in posterior cricoarytenoid expiratory duration and integrated inspiratory area were sleep state-dependent: area decreases were greatest in rapid eye movement sleep; expiratory duration increases were greatest in quiet sleep. Neural mechanisms underlying laryngeal abductor activity are sleep state-dependent and appear to be affected more than diaphragmatic mechanisms by baroreceptor stimulation.     

Not paralysis, but dystonia causes stridor in multiple system atrophy

Electromyography (EMG) was performed in 10 patients with multiple system atrophy, laryngeal or pharyngeal symptoms, or both. In patients with stridor, EMG during quiet breathing revealed persistent tonic activity in both abductor and adductor vocal cord muscles. In patients with dysphagia, the cricopharyngeal muscle showed persistent EMG activity throughout all phases of swallowing. Botulinum toxin injection into the adductor muscle determined subjective improvement and reduced tonic EMG activity. Therefore, the cause of stridor in multiple system atrophy is dystonia of the vocal cords.     

Laryngeal electromyography: an evidence-based review

This article reports on an evidence-based review of laryngeal electromyography (EMG) as a technique for use in the diagnosis, prognosis, and treatment of laryngeal movement disorders including the laryngeal dystonias, vocal fold paralysis, and other neurolaryngological disorders. The authors performed a systematic review of the medical literature from 1944 through 2001 on the clinical application of EMG to laryngeal disorders. Thirty-three of the 584 articles met the predefined inclusion criteria. The evidence demonstrated that in a double-blind treatment trial of botulinum toxin versus saline, laryngeal EMG used to guide injections into the thyroarytenoid muscle in persons with adductor spasmodic dysphonia was beneficial. A cross-over comparison between laryngeal EMG-guided injection and endoscopic injection of botulinum toxin into the posterior cricoarytenoid muscle in abductor spasmodic dysphonia found no significant difference between the two techniques and no significant treatment benefit. Based on the evidence, laryngeal EMG is possibly useful for the injection of botulinum toxin into the thyroarytenoid muscle in the treatment of adductor spasmodic dysphonia. There were no evidence-based data sufficient to support or refute the value of laryngeal EMG for the other uses investigated, although there is extensive anecdotal literature suggesting that it is useful for each of them. There is an urgent need for evidence-based research addressing the use of laryngeal EMG for other applications.     

Laryngeal electromyography with separated surface electrodes in patients with multiple system atrophy presenting with vocal cord paralysis

When recording the activity of the posterior cricoarytenoid muscle (PCA) with surface electrodes, there is contamination from the surrounding muscles such as the cricopharyngeal muscle. We therefore devised a new oesophageal catheter electrode of the separate type, having three individual surface electrodes for the PCA, cricopharyngeal muscle and diaphragm. The records obtained with this catheter demonstrated satisfactory separation between PCA and cricopharyngeal muscle activities. We used this catheter in patients with multiple system atrophy presenting with vocal cord paralysis, who were awake or asleep. There were two interesting electromyographical findings, which were inspiratory activity of the adductor muscle (the thyroarytenoid muscle) and fade-out of the abductor muscle, that is, PCA activity during sleep. Although vocal cord paralysis is one of the most serious life-threatening complications, the precise mechanism has not been clarified. We believe that our catheter may be useful in investigating the mechanism of vocal cord paralysis which could cause sudden death in neurodegenerative disorders, including multiple system atrophy.     

Cytoarchitectonic organization of laryngeal motoneurons within the nucleus ambiguus of the cat

The central distribution of laryngeal motoneurons was studied in the cat by retrograde axonal transport of horseradish peroxidase. The enzyme was injected selectively into the cricothyroid (CT), lateral cricoarytenoid and thyroarytenoid (LCA-TA), and posterior cricoarytenoid (PCA) muscles of the larynx with or without the previous sectioning of the left laryngeal recurrent nerve (LR) or the left superior laryngeal nerve (SL). The CT motoneurons appeared as a compact group of medium-size cells located in the rostral one-third of the nucleus ambiguus (nA). The LCA-TA motoneurons were found in the caudal two-thirds of the nA, constituting a loose group of large motoneurons. The PCA motoneurons were located throughout the whole extend of the nA, the cells being large in the caudal pole and smaller in the rostral one-third of nA. Laryngeal muscle innervation was exclusively of ipsilateral origin. Axonal projections in the brain stem were different depending on the nerve (LR or SL) by which the efferent fibers were sent.     

Efferent activity in the phrenic nerve during the startle reflex in chloralose-anesthetized cats

Reflex activity in the phrenic nerve was studied in chloralose anesthetized cats during development of somatic startle reflexes in limb and lower intercostal nerves. It was shown that the main component of this activity during low-threshold reflexes evoked by acoustic, tactile and low-threshold somatic afferent stimulation was depression of phrenic inspiratory activity. The following reflex discharges were prevalent components of phrenic responses to high-threshold afferent stimulation: early, propriospinal (intercostal-to-phrenic reflex) and late, suprasegmental ones. The latter were of two types: inspiratory (observed mainly during inspiration in about 75% of experiments) and expiratory (observed during expiration in 25% of experiments) which could be classified as "phrenic startle reflexes". Modulation of all responses during the respiratory cycle was described. Structural characteristics of reflex responses evoked in the phrenic nerve by stimulation of various respiratory and nonrespiratory bulbar sites as well as their respiratory modulation have been analyzed. Organization of possible neurophysiological mechanisms of phrenic responses during startle reflexes is discussed.     

The participation of bulbar "respiratory" and "nonrespiratory" neurons in the development of the expiration reflex

The patterns of impulse activity of medullar neurons were investigated in nembutal-anaesthetized cats during the expiration reflex elicited by electrical stimulation of the internal branch of superior laryngeal nerve. It was shown that low- and high-threshold superior laryngeal afferents caused excitatory reflex reactions of different complexity in significant number of reticular nonrespiratory neurons. Respiratory neurons exhibited systemic changes of spontaneous activity, but in 22.4 per cent of them reflex responses were recorded. These responses occurred during activation of low-threshold laryngeal afferents. Oligo- and polysynaptic excitatory connections of low-threshold laryngeal afferents are found with inspiratory beta neurons, P-cells and laryngeal motoneurons, but inhibitory ones-with inspiratory gamma neurons. Participation of investigated neurons in the mechanisms of inhibition of inspiration, vocal cords closure and rate of breathing adaptive decrease during expiration reflex is discussed.     

Location of motoneurons supplying the intrinsic laryngeal muscles of rats. Horseradish peroxidase and fluorescence double-labeling study

This paper describes a qualitative and quantitative investigation of the location of the motoneurons innervating the intrinsic laryngeal muscles of rats. Injections of horseradish peroxidase, Diamidino Yellow and True Blue were made either in one or, simultaneously, in three laryngeal muscles. Unlike those in cats and rabbits, the motoneurons that make up the nucleus ambiguus (NA) in rats are not arranged in two separate subgroups, that is one belonging to the cricothyroid (CT) motoneurons and the other to the rest of the intrinsic laryngeal motoneurons. Instead, a superimposition of CT and posterior cricoarytenoid (PCA) motoneurons was observed in the rostral third of the NA. Motoneurons innervating the PCA, thyroarytenoid (TA) and lateral cricoarytenoid (LCA) muscle overlap in the medial third of the NA. Finally, in the region of the NA caudal to the obex, the TA and LCA motoneurons also overlap. Labeled motoneurons were located in the ipsilateral side to the injected muscle in all cases.     

Spontaneous and reflexly evoked activity in pharyngeal, laryngeal, and phrenic motoneurons of cat

Experiments were performed on spontaneously breathing cats that were anesthetized with chloralose-urethane in order to study changing excitability in pharyngeal, laryngeal, and phrenic motoneurons. Pharyngeal constrictor motoneurons chiefly were spontaneously active during expiration; the stylopharyngeus and certain motoneurons serving the superior pharyngeal constrictors were inactive during either phase of tidal respiration. While confirming work of previous investigators that feeble glossopharyngeal (epipharyngeal branch) and superior laryngeal nerve stimulation transiently attenuated on-going phrenic nerve activity, such evoked afferent volleys excited laryngeal motoneurons independent of phases of respiration. On-going pharyngeal constrictor nerve activity (expiration) was attenuated for 20–50 msec by superior laryngeal nerve stimulation and for 100–300 msec by glosso-pharyngeal nerve stimulation. Intercostal nerve stimulation evoked occasional pharyngeal constrictor nerve discharges but more commonly attenuated its spontaneous nerve activity. Stylopharyngeus and an unknown portion of the superior pharyngeal constrictor motoneurons were excited by afferent volleys evoked in glossopharyngeal and superior laryngeal nerves. These aforementioned changes in excitability were unaffected by total neuromuscular paralysis while maintaining artificial respiration. It is concluded that certain pharyngeal constrictor motoneurons of the cat are spontaneously active during tidal expiration, are under central respiratory control, and that their intermittence along with those serving the diaphragm and larynx by modest afferent volleys in the ninth and tenth cranial nerves stage the act of swallow or expulsive reflexes, or both.     

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 CO2 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.     

Withdrawal reflexes in adductor muscles elicited by electrical and magnetic stimulation of the obturator nerve

The withdrawal reflex in the short head of the biceps femoris muscle after electrical stimulation of the sural nerve at the ankle has been investigated in numerous studies. These studies have described two distinct responses: early (R-II) and late (R-III). However, withdrawal reflex activity of the adductor muscles in the legs has not been studied systematically. Adductor muscle reflex activity is important because it can produce serious clinical problems, such as adductor spasticity and spasms, during bladder surgery. The present study examined withdrawal reflex features of adductor muscles obtained by electrical and magnetic stimulation of the obturator nerve (ON) in 34 normal healthy subjects. Early adductor muscle withdrawal reflex responses were elicited by ipsilateral ON electrical stimulation with a mean latency of 45.7+/-2.0 ms (responses in 94% of subjects). Reflex responses were also obtained using magnetic stimulation at a similar incidence rate. Contralateral ON electrical stimulation resulted in a similar reflex, but with a lower incidence. ON and femoral nerve electrical and magnetic coil stimulation produced similar low-incidence responses in the vastus medialis. These findings indicate that short latency adductor withdrawal reflexes are easily obtained on both sides following electrical or magnetic stimulation of the ON, and they can be elicited by both nociceptive and nonnociceptive stimuli. These reflexes prepare the body for a proper response to incoming signals and likely serve to protect the pelvic floor and pelvic organs.