Stimulation Of Laryngeal Afferents Research Articles

Laryngeal Adductor Reflex Motor Bursts Rapidly Oscillate in the Cat

The laryngeal adductor reflex (LAR) is a protective behavior induced by stimulation of upper airway afferents. Laryngeal afferent information reaches interneurons in the medulla via the superior laryngeal nerve (SLN). Expiratory laryngeal motoneurons (ELMs) are subsequently excited causing contraction of the thyroarytenoid (TA) and posterior cricoarytenoid (PCA) muscles. The LAR consists of a short latency (~10 ms), short duration (~10 ms) burst (R1) and a longer latency (~40ms) burst (R2) in laryngeal electromyogram (EMG) activity. We have shown that R1 LAR magnitude undergoes frequency‐dependent depression in response to electrical stimulation (eStim) of the SLN. Based on preliminary observations, we hypothesized that the R1 component of the LAR induced by SLN eStim oscillates. In anesthetized, spontaneously breathing cats (n=12: 7 males, 5 females), we recorded EMG activity from the TA bilaterally and the PCA unilaterally. The SLN was electrically stimulated at 2 or 20 Hz to evoke TA LAR ipsilateral to stimulation (iTA). No sex differences in TA or PCA LAR EMG magnitude were observed. Consistent with previous findings, TA LAR magnitude differed with breathing phase; normalized iTA LAR magnitude was 10% larger during expiration compared to inspiration. TA LAR contralateral to stimulation (cTA) and ipsilateral PCA LAR magnitudes were not breathing phase dependent. TA and PCA LAR EMG oscillations were highly variable across animals (range: 244 to 1140 Hz). Mean iTA EMG frequency was decreased at 20 Hz (2 to 20 Hz: 616 to 518 Hz, p=0.05). Mean cTA periodicity trended down at 20 Hz (2 to 20 Hz cTA: 647 to 531 Hz, p=0.07). In separate decerebrate, paralyzed cats (n=13), extracellular recordings of neurons in the ventral respiratory column (VRC) with multiple electrode arrays were conducted. Peristimulus time histograms provided evidence that ELMs oscillated at frequencies following SLN eStim similar to the lower range of periodicities in TA EMGs in unparalyzed animals. A subset of neurons recorded in the VRC were excited or inhibited by SLN eStim at short latencies. Spike train analysis suggested that ELM oscillations could be accounted for by synaptic inhibition and excitation from laryngeal‐responsive VRC inspiratory, expiratory, and non‐breathing modulated neurons. We conclude that the LAR is temporally regulated by a complex, anatomically distributed brainstem network that is capable of strong synchrony in response to stimulation of laryngeal afferents. We speculate that the function of this laryngeal‐responsive network is to exert tight regulatory control over LAR patterning.Support or Funding InformationSupported by NIH HL131716 and 3OT2OD023854

Selective suppression of late laryngeal adductor responses by N-methyl-D-aspartate receptor blockade in the cat.

Laryngeal adductor responses to afferent stimulation play a key role in airway protection. Although vital for protection during cough and swallow, these responses also must be centrally controlled to prevent airway obstruction by laryngospasm during prolonged stimulation. Our purpose was to determine the role of N-methyl-D-aspartate (NMDA) receptors in modulating early R1 responses (at 9 ms) and/or later more prolonged R2 responses (at 36 ms) during electrical stimulation of the laryngeal afferent fibers contained in the internal branch of the superior laryngeal nerve in the cat. The percent occurrence, amplitude, and conditioning of muscle responses to single superior laryngeal nerve (SLN) stimuli presented in pairs at interstimulus intervals of 250 ms were measured in three experiments: 1) animals that had ketamine as anesthetic premedication were compared with those who did not, when both were maintained under alpha-chloralose anesthesia. 2) The effects of administering ketamine in one group of animals were compared with increasing the depth of alpha-chloralose anesthesia without NMDA receptor blockade in another group of animals. 3) The effects of dextromethorphan (without anesthetic effects) were examined in another group of animals. In the first experiment, the occurrence of R2 responses were reduced from 95% in animals without ketamine premedication to 25% in animals with ketamine premedication (P = 0.015). No differences occurred in the occurrence, amplitude, latency, or conditioning effects on R1 responses between these groups. In the second experiment, the occurrence of R2 responses was reduced from 96 to 79% after an increase in the depth of anesthesia with alpha-chloralose in contrast with reductions in R2 occurrence from 98 to 19% following the administration of ketamine to induce NMDA receptor blockade along with increased anesthesia (P = 0.025). In the third experiment, R2 occurrence was reduced from 89 to 27% (P = 0.017) with administration of dextromethorphan while R1 response occurrence and amplitude did not change. In each of these experiments, NMDA receptor blockade did not have significant effects on cardiac or respiratory rates in any of the animals. The results demonstrate that NMDA receptors play an essential role in long latency R2 laryngeal responses to laryngeal afferent stimulation. On the other hand, early R1 laryngeal adductor responses are likely to involve non-NMDA receptor activation.

Laryngeal afferent stimulation enhances fos immunoreactivity in periaqueductal gray in the cat

The main functions of the larynx are protection of the airways, respiration, and vocalization. Previous studies have suggested a link between the mechanisms controlling vocalization and afferent feedback from the larynx. We inquired whether stimulation of the laryngeal afferents that run in the internal branch of the superior laryngeal nerve (ISLN) activates neurons of the periaqueductal gray (PAG), a midbrain region implicated in vocalization. We counted the number of neurons expressing Fos, the protein product of the immediate early gene c-fos, in the PAG. The counts were done both in experimental cats after electrical stimulation of the ISLN and nonstimulated controls. We also investigated the possible presence of nitric oxide synthase, an enzyme that synthesizes nitric oxide, in PAG neurons that respond to laryngeal afferent stimulation by double labeling for reduced nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase and Fos. Fos expression was significantly greater (P < or = 0.00714) in the lateral and dorsolateral regions of the PAG in the experimental group than in the controls. The Fos-immunoreactive neurons did not contain NADPH-diaphorase, a marker for nitric oxide synthase. Our study suggests that laryngeal afferent stimulation activates neurons in discrete longitudinal columns of the PAG including the regions that have previously been shown to be involved in vocalization, and that these neurons do not contain nitric oxide synthase.

Laryngeal afferent stimulation enhances fos immunoreactivity in periaqueductal gray in the cat This article is a US Government work and, as such, is in the public domain in the United States of America.

Laryngeal afferent stimulation enhances fos immunoreactivity in periaqueductal gray in the cat This article is a US Government work and, as such, is in the public domain in the United States of America.

Laryngeal afferent inputs to the nucleus of the solitary tract.

The following study was undertaken to examine the integration of laryngeal afferent inputs within the nucleus of the solitary tract (NTS), the primary site of termination of laryngeal afferent fibers. Intracellular recordings were obtained from 63 cells that responded to electrical stimulation of the superior laryngeal nerve (SLN) with an excitatory postsynaptic potential (EPSP; n = 49), an excitatory-inhibitory postsynaptic potential (EPSP-IPSP) sequence (n = 13), or an IPSP (n = 1). Mechanical stimulation of laryngeal mechanoreceptors revealed a variety of response patterns (e.g., slowly and rapidly adapting depolarizations or hyperpolarizations). Two types of response to increasing SLN stimulus frequency were observed. In 11 cells SLN-evoked EPSP amplitude at 10 Hz was only 47 +/- 4% of the amplitude at 1 Hz, while in 6 cells EPSP amplitude at 10 Hz was virtually identical (93 +/- 3%) to that at 1 Hz. Time-dependent inhibitory interactions occurred between SLN inputs to NTS neurons at intervals between 50 and 400 ms and in the absence of any change in membrane potential. NTS neuronal responses to brief activation of laryngeal mechanoreceptors correspond well to discharge patterns described for individual laryngeal mechanoreceptors. Frequency-dependent filtering and time-dependent inhibitory interactions might modify NTS neuronal responses during more intense stimulation of laryngeal afferents.

Synaptic inputs to medullary respiratory neurons from superior laryngeal afferents in the cat

Synaptic inputs from afferents in the superior laryngeal nerve (SLN) to medullary respiratory neurons ( n=154) in the dorsal respiratory group (DRG), ventral respiratory group (VRG) and the region of the Bo¨tzinger complex (BOT) were studied in anesthetized cats. Single pulse stimulation of the SLN-evoked monosynaptic EPSPs in most inspiratory bulbospinal (I-BS) neurons in the DRG, and disynaptic or oligosynaptic chloride-dependent IPSPs in other I-BS neurons in the DRG and VRG. Stimulation of laryngeal afferents also inhibited oligosynaptically expiratory bulbospinal neurons in the VRG, and all types of respiratory neurons recorded in the BOT region. Oligosynaptic potentials (usually EPSPs) were recorded in inspiratory and expiratory laryngeal motoneurons. These results provide evidence of a processing of SLN-evoked synaptic responses by all tested groups of medullary respiratory neurons. The pathways mediating these synaptic responses are discussed.

Antagonistic interaction of laryngeal and central chemoreceptor respiratory reflexes.

Stimulation of laryngeal afferent fibers evokes a profound reflex inhibition of central respiratory drive. The interaction of this airway reflex with chemoreceptive ventilatory control mechanisms is poorly understood. The present study was undertaken to determine whether there is significant interaction between the effects of central chemoreceptor and laryngeal afferent stimulation on central inspiratory activity and, if so, to also determine the nature of the interaction. The effect of electrical stimulation of the superior laryngeal nerve (SLN) on the timing and intensity of central inspiratory activity was determined from the rectified and filtered phrenic neurogram in 10 dogs. Each dogs was decerebrated, artificially ventilated, vagotomized, and had the carotid bodies denervated. In each case, stimulation of the right SLN at 3 and 10 Hz caused a frequency-dependent slowing or arrest of central inspiratory activity. Increases in arterial PCO2 (PaCO2) attenuated the absolute level of inhibition of central inspiratory activity recorded during both SLN stimulation and control periods. Tp clarify the nature of the interaction between chemoreceptor and laryngeal afferent stimulation, the relationship between PaCO2 and central inspiratory activity was investigated during stimulation of the SLN at 0, 3, and 10 Hz. Control central inspiratory activity increased as a sigmoidal function of PaCO2. This sigmoidal relationship was greatly depressed during SLN stimulation but did not appear to be shifted along the PaCO2 axis. The results of this study therefore suggest that the interaction between central chemoreceptor and laryngeal afferent stimulation is multiplicative: the inhibition of the central inspiratory activity is mediated by an attenuation and not a resetting of central chemoreflexes.

Inhibition of cardiac sympathetic nerve activity during swallowing evoked by laryngeal afferent stimulation in the cat

Spontaneous swallowing or the buccopharyngeal phase of swallowing evoked by electrical stimulation of the superior laryngeal nerve was accompanied by a pronounced decrease of sympathetic activity in the cardiac nerve. This reduction of sympathetic activity was not related to other influences such as postinspiratory inhibition or baroreceptor-mediated inhibition. Intraceullular recordings from sympathetic preganglionic neurones revealed hyperpolarization during the buccopharyngeal phase of swallowing, possibly due to postsynaptic inhibition.

Reflex bradypnea elicited by cigarette smoke inhaled through an isolated larynx

The respiratory reflex responses elicited by laryngeal exposure to cigarette smoke were studied in 23 chloralose anesthetized dogs. A balloon-in-box system was connected to the breathing circuit, which allowed smoke to be inhaled spontaneously through an isolated larynx while preserving its normal respiratory flow and pressure. Our results in this study showed the following. (1) Two tidal breaths of cigarette smoke inhaled through the larynx triggered a mild but consistent bradypnea: expiratory duration (T E) increased from a control of 3.13 ± 0.18 sec (mean ± SEM) to peak of 4.07 ± 0.24 sec. during smoke inhalation. The slowing of respiration occurred only during the period of smoke inhalation and returned quickly toward control after resuming air breathing. (2) No concomitant cardiovascular response was detected in these animals. (3) There was no significant difference in the prolongation of T E between responses to low- and high-nicotine cigarette smoke. (4) The bradypneic response to smoke was completely abolished by application of topical anesthetic (lidocaine hydrochloride) to the mucosa of this airway segment. These results suggest that the smoke-induced reflex bradypnea is probably elicited by a stimulation of laryngeal afferents.

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