Jan Jakus

Neurogenesis of cough, other airway defensive behaviors and breathing: A holarchical system?

Cough and breathing are generated by a common muscular system. However, these two behaviors differ significantly in their mechanical features and regulation. The current conceptualization of the neurogenic mechanism for these behaviors holds that the multifunctional respiratory pattern generator undergoes reconfiguration to produce cough. Our previous results indicate the presence of a functional cough gate mechanism that controls the excitability of this airway defensive behavior, but is not involved in the regulation of breathing. We propose that the neurogenesis of cough, breathing, and other nonbreathing behaviors is controlled by a larger network, of which the respiratory pattern generator is part. This network we term a holarchical system. This system is governed by functional control elements known as holons, which confer unique regulatory features to each behavior. The cough gate is an example of such a holon. Neurons that participate in a cough holon may include behavior selective elements. That is, neurons that are either specifically recruited during cough and/or tonically-active neurons with little or no modulation during breathing but with significant alterations in discharge during coughing. We also propose that the holarchical system is responsible for the orderly expression of different airway defensive behaviors such that each motor task is executed in a temporally and mechanically discrete manner. We further propose that a holon controlling one airway defensive behavior can regulate the excitability of, and cooperate with, holons unique to other behaviors. As such, co-expression of multiple rhythmic behaviors such as cough and swallow can occur without compromising airway defense.

Brainstem circuitry of tracheal-bronchial cough: c-fos study in anesthetized cats.

The c-fos gene expression method was used to localize brainstem neurons functionally related to the tracheal-bronchial cough on 13 spontaneously breathing, pentobarbitone anesthetized cats. The level of Fos-like immunoreactivity (FLI) in 6 animals with repetitive coughs (170+/-12) induced by mechanical stimulation of the tracheobronchial mucosa was compared to FLI in 7 control non-stimulated cats. Thirty-four nuclei were compared for the number of labeled cells. Enhanced cough FLI was found bilaterally at following brainstem structures, as compared to controls: In the medulla, FLI was increased in the medial, interstitial and ventrolateral subnuclei of the solitary tract (p < 0.02), in the retroambigual nucleus of the caudal medulla (p < 0.05), in the ambigual, paraambigual and retrofacial nuclei of the rostral medulla along with the lateral reticular nuclei, the ventrolateral reticular tegmental field (p < 0.05), and the raphe nuclei (p < 0.05). In pons, increased FLI was detected in the lateral parabrachial and Kölliker-Fuse nuclei (p < 0.01), in the posteroventral cochlear nuclei (p < 0.01), and the raphe midline (p < 0.05). Within the mesencephalon cough-related FLI was enhanced at the rostral midline area (p < 0.05), but a decrease was found at its caudal part in the periaqueductal gray (p < 0.02). Results of this study suggest a large medullary - pontine - mesencephalic neuronal circuit involved in the control of the tracheal-bronchial cough in cats.

Short reflex expirations (expiration reflexes) induced by mechanical stimulation of the trachea in anesthetized cats.

Fifty spontaneously breathing pentobarbital-anesthetized cats were used to determine the incidence rate and parameters of short reflex expirations induced by mechanical stimulation of the tracheal mucosa (ERt). The mechanical stimuli evoked coughs; in addition, 67.6% of the stimulation trials began with ERt. The expiration reflex mechanically induced from the glottis (ERg) was also analyzed (99.5% incidence, p < 0.001 compared to the incidence of ERt). We found that the amplitudes of abdominal, laryngeal abductor posterior cricoarytenoid, and laryngeal adductor thyroarytenoid electromyograms (EMG) were significantly enhanced in ERg relative to ERt. Peak intrathoracic pressure (esophageal or intra-pleural pressure) was higher during ERg than ERt. The interval between the peak in EMG activity of the posterior cricoarytenoid muscle and that of the EMG of abdominal muscles was lower in ERt compared to ERg. The duration of thyroarytenoid EMG activity associated with ERt was shorter than that in ERg. All other temporal features of the pattern of abdominal, posterior cricoarytenoid, and thyroarytenoid muscles EMGs were equivalent in ERt and ERg.In an additional 8 cats, the effect of codeine administered via the vertebral artery was tested. Codeine, in a dose (0.03 mg/kg) that markedly suppressed cough did not significantly alter either the incidence rate or magnitudes of ERt.In the anesthetized cat the ERt induced by mechanical stimulation of the trachea was similar to the ERg from the glottis. These two reflex responses differ substantially only in the frequency of occurrence in response to mechanical stimulus and in the intensity of motor output.

Multifunctional ventral respiratory group: Bulbospinal expiratory neurons play a role in pudendal discharge during vomiting

Pudendal motoneurons are activated in phasic bursts during the retching and expulsion phases of vomiting. The resulting contraction of the anal and urethral sphincters serves to maintain continence during the large increase in abdominal pressure that occurs during vomiting. We evaluated the contribution of bulbospinal expiratory neurons located in the portion of the ventral respiratory group (VRG) caudal to the obex (nucleus retroambigualis) to the control of pudendal motoneurons during fictive vomiting in decerebrate, paralyzed cats. Pudendal nerve discharge is abolished by cutting the axons of caudal VRG expiratory neurons as they cross the midline between the obex and C1 before descending in the spinal cord. All caudal VRG expiratory neurons that were antidromically activated from the sacral spinal cord, where the pudendal motor pool (nucleus of Onuf) is located, discharged strongly during the end of the expulsion phase of vomiting. However, only a small proportion of these neurons was active in phase with pudendal discharge during the retching phase. The apparent involvement of caudal VRG expiratory neurons in the control of pudendal motoneurons during vomiting is another example of the multifunctional role that can be played by respiratory-related neurons in the mammalian nervous system.

Central antitussive effect of codeine in the anesthetized rabbit

BackgroundCodeine represents a commonly used drug to suppress cough. Central antitussive effect of codeine has been confirmed in a number of animal studies. However, available data related to antitussive activity of codeine in rabbits are very limited.ObjectiveWe investigated the effects of codeine on cough, single expiratory responses (expiration-like reflex) induced by mechanical tracheo-bronchial stimulation, and on the sneeze reflex in the anesthetized rabbit.Materials and methodsTwenty pentobarbitone anesthetized spontaneously breathing rabbits were used for the study. Increasing doses of codeine (codeinum dihydrogenphosphate, Interpharm) were injected intravenously (iv); 0, 0.15, 0.76, and 3.78 mg/kg of codeine dissolved in saline, 0.25 ml/kg) or intracerebroventricularly (icv); 0, 0.015, 0.076, and 0.378 mg/kg of codeine dissolved in artificial cerebrospinal fluid, 0.033 ml/kg.ResultsBoth iv and icv injections of codeine led to a dose-dependent reduction of coughing provoked by tracheo-bronchial stimulation; however, the doses differed substantially. The effective cumulative dose for a 50% reduction in the number of coughs was 3.9 and 0.11 mg/kg after iv and icv administration of codeine, respectively; representing about 35-fold higher efficacy of the icv route. The numbers of expiration-like responses and sneeze reflex responses remained unchanged.ConclusionsThe study confirmed the central antitussive effect of codeine, but showed a low sensitivity of sneeze and expiration reflex to codeine. We validated the experimental model of an anesthetized rabbit for studies on central antitussive action.

Intranasal TRPV1 agonist capsaicin challenge and its effect on c-fos expression in the guinea pig brainstem.

Central neuronal interaction seems to play a role in pathogenesis of upper airway cough syndrome. In the guinea pig model we used the method c-fos expression to identify neurons involved in processing of nociceptive nasal stimuli and their contribution to enhancement of cough. 21 spontaneously breathing, urethane anaesthetized animals were used. The controls received intranasal saline, stimulation group received capsaicin (15 microl, 50 microM), and not-treated group was free of nasal challenge. After 2 h animals were deeply anaesthetized, exsanguinated and transcardially perfused with saline and paraformaldehyde. The brainstems were removed, post-fixed, and slices were processed immunohistochemically for c-fos. In capsaicin group the FLI was detected in the nTs 0.5 mm caudal, 1.5 mm lateral to the obex, the area postrema, LRN and VRG. Intensive FLI was identified in trigeminal nuclear complex. Mean number of FOS positive neurons per section was significantly higher in capsaicin group than that in no-treatment controls or saline controls at the level of obex (p<0.01). Neurons of nTs and VRG clearly activated after nasal provocation may participate in enhancement of cough.

Brainstem regions involved in the expiration reflex. A c-fos study in anesthetized cats.

Expression of the immediate-early gene c-fos, a marker of neuronal activation, was employed to localize brainstem neuronal populations functionally related to the expiration reflex (ER). Twelve spontaneously breathing, non-decerebrate, pentobarbital anesthetized cats were used. The level of Fos-like immunoreactivity (FLI) in 6 animals with repetitive ERs mechanically induced from the glottis (296+/-9 ERs) was compared to FLI in 6 control non-stimulated cats. Respiratory rate, arterial blood pressure, and end tidal CO(2) concentration remained stable during the experiment. In the medulla, increased FLI was found in the region of nucleus tractus solitarii (p<0.001), in the ventrolateral medulla along with the lateral tegmental field (p<0.01), and in the vestibular nuclei (p<0.01). In the pons, increased FLI was detected in the caudal extensions of the lateral parabrachial and Kölliker-Fuse nuclei (p<0.05). Within the rostral mesencephalon, FLI was enhanced in the midline area (p<0.05). A lower level of ER-related FLI compared to control animals was detected in the pontine raphe region (p<0.05) and the lateral division of mesencephalic periaqueductal gray (p<0.05). The results suggest that the ER is coordinated by a complex long loop of medullary-pontine-mesencephalic neuronal circuits, some of which may differ from those of other respiratory reflexes. The FLI related to the expulsive behavior ER differs from that induced by laryngeal stimulation and laryngeal adductor responses, particularly in ventrolateral medulla and mesencephalon.

The course of lung inflation alters the central pattern of tracheobronchial cough in cat-The evidence for volume feedback during cough.

The effect of volume-related feedback and output airflow resistance on the cough motor pattern was studied in 17 pentobarbital anesthetized spontaneously-breathing cats. Lung inflation during tracheobronchial cough was ventilator controlled and triggered by the diaphragm electromyographic (EMG) signal. Altered lung inflations during cough resulted in modified cough motor drive and temporal features of coughing. When tidal volume was delivered (via the ventilator) there was a significant increase in the inspiratory and expiratory cough drive (esophageal pressures and EMG amplitudes), inspiratory phase duration (CTI), total cough cycle duration, and the duration of all cough related EMGs (Tactive). When the cough volume was delivered (via the ventilator) during the first half of inspiratory period (at CTI/2-early over inflation), there was a significant reduction in the inspiratory and expiratory EMG amplitude, peak inspiratory esophageal pressure, CTI, and the overlap between inspiratory and expiratory EMG activity. Additionally, there was significant increase in the interval between the maximum inspiratory and expiratory EMG activity and the active portion of the expiratory phase (CTE1). Control inflations coughs and control coughs with additional expiratory resistance had increased maximum expiratory esophageal pressure and prolonged CTE1, the duration of cough abdominal activity, and Tactive. There was no significant difference in control coughing and/or control coughing when sham ventilation was employed. In conclusion, modified lung inflations during coughing and/or additional expiratory airflow resistance altered the spatio-temporal features of cough motor pattern via the volume related feedback mechanism similar to that in breathing.

Plasticity of emesis to a 5-HT3 agonist: effect of order of visceral nerve cuts.

Serotonin type 3 receptors are critical for vomiting induced by radiation and cancer chemotherapy. To investigate the mechanisms involved, we analyzed the effects of abdominal visceral denervation using cats. Thresholds for vomiting induced by the serotonin-3 receptor agonist phenylbiguanide varied depending on the order of chronic nerve transections. Lower thoracic vagotomy, splanchnicectomy, and their sequential combination increased thresholds. However, when splanchnicectomy, which causes a large increase in threshold, was followed 14-51 days later by vagotomy, thresholds dropped by an average of 40%. These results demonstrate plasticity in, and potential for modulation of, the emetic reflex.

Resuscitation and auto resuscitation by airway reflexes in animals.

Various diseases often result in decompensation requiring resuscitation. In infantsmoderate hypoxia evokes a compensatory augmented breath – sigh and more severehypoxia results in a solitary gasp. Progressive asphyxia provokes gasping respirationsaving the healthy infant – autoresuscitation by gasping. A neonate with suddeninfant death syndrome, however, usually will not survive. Our systematic research inanimals indicated that airway reflexes have similar resuscitation potential asgasping respiration. Nasopharyngeal stimulation in cats and most mammals evokes theaspiration reflex, characterized by spasmodic inspiration followed by passiveexpiration. On the contrary, expiration reflex from the larynx, or cough reflex fromthe pharynx and lower airways manifest by a forced expiration, which in cough ispreceded by deep inspiration. These reflexes of distinct character activate thebrainstem rhythm generators for inspiration and expiration strongly, but differently.They secondarily modulate the control mechanisms of various vital functions of theorganism. During severe asphyxia the progressive respiratory insufficiency may inducea life-threatening cardio-respiratory failure. The sniff- and gasp-like aspirationreflex and similar spasmodic inspirations, accompanied by strong sympatho-adrenergicactivation, can interrupt a severe asphyxia and reverse the developing dangerouscardiovascular and vasomotor dysfunctions, threatening with imminent loss ofconsciousness and death. During progressive asphyxia the reversal of graduallydeveloping bradycardia and excessive hypotension by airway reflexes starts withreflex tachycardia and vasoconstriction, resulting in prompt hypertensive reaction,followed by renewal of cortical activity and gradual normalization of breathing. Acombination of the aspiration reflex supporting venous return and the expiration orcough reflex increasing the cerebral perfusion by strong expirations, provides apowerful resuscitation and autoresuscitation potential, proved in animal experiments.They represent a simple but unique model tested in animal experiments.