Plasticity of central mechanisms for cough.

Cough is associated with plasticity of putative cough afferent fibres, but whether plasticity in the brainstem network contributes is less well understood. A key site in the CNS network is the nucleus tractus solitarius (NTS), the first synaptic contact of the primary afferent fibres. We sought to develop a conscious guinea pig model to detect enhanced cough, to focus on the NTS as a potential site for plasticity, and to test a role for substance P in the NTS since the neuropeptide has been implicated in plasticity of the vagal afferent fibres. Guinea pigs were exposed to second-hand tobacco smoke (SHS) or filtered air (FA) from 1-6 weeks of age. At 5 weeks, cannulae were implanted in the NTS. At 6 weeks, either vehicle or a neurokinin 1 (NK-1) receptor antagonist was injected into the NTS of the conscious guinea pigs who were then exposed to citric acid aerosol. SHS exposure significantly enhanced citric acid-induced cough (56%, P<0.05), an effect attenuated by NTS NK-1 receptor blockade (P<0.05). The findings suggest that one possible mechanism for plasticity in cough is related to substance P effects in the NTS. Future studies will be required to investigate the possible mechanisms underlying the role of substance P as well as other mechanisms in generating SHS-induced cough.

Frequency response functions and information capacities of paired spider mechanoreceptor neurons.

Pseudorandom white-noise stimulation followed by direct spectral estimation was used to obtain linear frequency response and coherence functions from paired, but dynamically different, spider mechanosensory neurons. The dynamic properties of the two neuron types were similar with either mechanical or electrical stimulation, showing that action potential encoding dominates the dynamics. Phase-lag data indicated that action potential initiation occurs more rapidly during mechanical stimulation, probably in the distal sensory dendrites. Total information capacity, calculated from coherence, as well as information per action potential, were both similar in the two types of neurons, and similar to the few available estimates from other spiking neurons. However, information capacity and information per action potential both depended strongly on neuronal firing rate, which has not been reported before.

Inactivation of voltage-activated Na(+) currents contributes to different adaptation properties of paired mechanosensory neurons.

Voltage-activated sodium current (I(Na)) is primarily responsible for the leading edge of the action potential in many neurons. While I(Na) generally activates rapidly when a neuron is depolarized, its inactivation properties differ significantly between different neurons and even within one neuron, where I(Na) often has slowly and rapidly inactivating components. I(Na) inactivation has been suggested to regulate action potential firing frequency in some cells, but no clear picture of this relationship has emerged. We studied I(Na) in both members of the paired mechanosensory neurons of a spider slit-sense organ, where one neuron adapts rapidly (type A) and the other slowly (type B) in response to a step depolarization. In both neuron types I(Na) activated and inactivated with single time constants of 2--3 ms and 5--10 ms, respectively, varying with the stimulus intensity. However, there was a clear difference in the steady-state inactivation properties of the two neuron types, with the half-maximal inactivation (V(50)) being -40.1 mV in type A neurons and -58.1 mV in type B neurons. Therefore I(Na) inactivated closer to the resting potential in the more slowly adapting neurons. I(Na) also recovered from inactivation significantly faster in type B than type A neurons, and the recovery was dependent on conditioning voltage. These results suggest that while the rate of I(Na) inactivation is not responsible for the difference in the adaptation behavior of these two neuron types, the rate of recovery from inactivation may play a major role. Inactivation at lower potentials could therefore be crucial for more rapid recovery.

Predicting the responses of mechanoreceptor neurons to physiological inputs by nonlinear system identification.

The nonlinear dynamic properties of action potential encoding were studied in mechanosensory neurons innervating the slits of a slit-sense organ in the tropical wandering spider, Cupiennius salei. The organ contains two types of neurons that are morphologically similar but have different dynamic properties. Type A neurons produce only one or two action potentials in response to a mechanical or electrical stimulus of any suprathreshold amplitude, while type B neurons can fire prolonged bursts of action potentials in response to similar stimuli. Neurons were stimulated with pseudorandomly modulated intracellular current while recording the resultant fluctuations in membrane potential and action potentials. A parallel cascade method was used to estimate a third-order Volterra series to describe the nonlinear dynamic relationship between membrane potential and action potentials. Kernels measured for the two types of neurons had reproducible forms that showed differences between the two neuron types. The measured kernels were able to predict the responses of the neurons to novel pseudorandomly modulated inputs with reasonable fidelity. However, the Volterra series did not adequately predict the difference in responses to step depolarizations.

Low-voltage-activated calcium current does not regulate the firing behavior in paired mechanosensory neurons with different adaptation properties.

Low-voltage-activated Ca(2+) currents (LVA-I(Ca)) are believed to perform several roles in neurons such as lowering the threshold for action potentials, promoting burst firing and oscillatory behavior, and enhancing synaptic excitation. They also may allow rapid increases in intracellular Ca(2+) concentration. We discovered LVA-I(Ca) in both members of paired mechanoreceptor neurons in a spider, where one neuron adapts rapidly (Type A) and the other slowly (Type B) in response to a step stimulus. To learn if I(Ca) contributed to the difference in adaptation behavior, we studied the kinetics of I(Ca) from isolated somata under single-electrode voltage-clamp and tested its physiological function under current clamp. LVA-I(Ca) was large enough to fire single action potentials when all other voltage-activated currents were blocked, but we found no evidence that it regulated firing behavior. LVA-I(Ca) did not lower the action potential threshold or affect firing frequency. Previous experiments have failed to find Ca(2+)-activated K(+) current (I(K(Ca))) in the somata of these neurons, so it is also unlikely that LVA-I(Ca) interacts with I(K(Ca)) to produce oscillatory behavior. We conclude that LVA-Ca(2+) channels in the somata, and possible in the dendrites, of these neurons open in response to the depolarization caused by receptor current and by the voltage-activated Na(+) current (I(Na)) that produces action potential(s). However, the role of the increased intracellular Ca(2+) concentration in neuronal function remains enigmatic.

Voltage-activated potassium outward currents in two types of spider mechanoreceptor neurons.

We studied the properties of voltage-activated outward currents in two types of spider cuticular mechanoreceptor neurons to learn if these currents contribute to the differences in their adaptation properties. Both types of neurons adapt rapidly to sustained stimuli, but type A neurons usually only fire one or two action potentials, whereas type B neurons can fire bursts lasting several hundred milliseconds. We found that both neurons had two outward current components, 1) a transient current that activated rapidly when stimulated from resting potential and inactivated with maintained stimuli and 2) a noninactivating outward current. The transient outward current could be blocked by 5 mM tetraethylammonium chloride, 5 mM 4-aminopyridine, or 100 microM quinidine, but these blockers also reduced the amplitude of the noninactivating outward current. Charybdotoxin or apamin did not have any effect on the outward currents, indicating that Ca2+-activated K+ currents were not present or not inhibited by these toxins. The only significant differences between type A and type B neurons were found in the half-maximal activation (V50) values of both currents. The transient current had a V50 value of 9. 6 mV in type A neurons and -13.1 mV in type B neurons, whereas the V50 values of noninactivating outward currents were -48.9 mV for type A neurons and -56.7 mV for type B neurons. We conclude that, although differences in the activation kinetics of the voltage-activated K+ currents could contribute to the difference in the adaptation behavior of type A and type B neurons, they are not major factors.

Vagal esophageal receptors in anesthetized dogs: mechanical and chemical responsiveness.

This study was performed to evaluate the characteristics of esophageal receptors in anesthetized and artificially ventilated dogs. The electrical activity of the esophageal afferents was recorded from the peripheral cut end of the cervical vagus nerve. A cuffed catheter was inserted into the esophagus at the level of the third tracheal ring and was used to establish the esophageal location of the endings. Most of the receptors were localized in the intrathoracic portion of the esophagus. The majority of the receptors studied (36 of 43) showed a slow adaptation to a maintained stretch of the esophageal wall. Vagal cooling blocked receptor activity at temperatures ranging from 3.5 to 25 degrees C. Twenty-eight of 43 receptors, including 4 rapidly adapting endings (RAR), were challenged with saline, HCl + pepsin (HCl-P; pH 1) and distilled water (8 ml, 37 degrees C). HCl-P solutions specifically stimulated only three receptors; saline or water did not. Five slowly adapting receptors and two RARs were also challenged with topically applied capsaicin; only one RAR was stimulated. To ascertain a possible effect of smooth muscle contraction, 17 receptors were tested with intravenous injections of ACh and/or asphyxia; only 4 were stimulated. These characteristics do not support an important reflexogenic role of the esophagus in response to chemical stimuli.

Larynx vs. esophagus as reflexogenic sites for acid-induced bronchoconstriction in dogs.

Bronchoconstriction in asthmatic patients is frequently associated with gastroesophageal reflux. However, it is still unclear whether bronchoconstriction originates from the esophagus or from aspiration of the refluxate into the larynx and larger airway. We compared the effect of repeated esophageal and laryngeal instillations of HCl-pepsin (pH 1.0) on tracheal smooth muscle activity in eight anesthetized and artificially ventilated dogs. Saline was used as control. We used pressure in the cuff of an endotracheal tube (Pcuff) as a direct index of smooth muscle activity at the level of the larger airways controlled by vagal efferents. The Pcuff values of the first 60 s after instillations were averaged, and the difference from the baseline values was evaluated. Changes in Pcuff were significantly greater with laryngeal than with esophageal instillations (P = 0.0166). HCl-pepsin instillation into the larynx evoked greater responses than did saline (P = 0.00543), whereas no differences were detected with esophageal instillations. Repeated laryngeal exposure enhanced the responsiveness significantly (P < 0. 001). Our data indicate that the larynx is more important than the esophagus as a reflexogenic site for the elicitation of reflex bronchoconstriction in response to acidic solutions.

Does histamine stimulate trigeminal nasal afferents?

The response to histamine of nasal afferents has been studied in guinea pigs by recording the electrical activity of the whole ethmoidal nerve (EN) or that of single units. Guinea pigs were anaesthetized with urethane and breathed through a tracheostomy. Prior to intranasal instillation of histamine (1 x 10(-4)-10(-1) M), the nasal mucosa was treated with 20 microl of saline (0.9% NaCl) or HCl (pH = 2), and in some cases, H2SO4 (pH = 2). In other experiments, following HCl instillation animals were pretreated by tripelennamine (1 x 10(-2) M) and/or cimetidine (1 x 10(-2) M) in order to determine the histamine receptor type of sensory nerve endings. Whole EN activity was not stimulated even by the highest dose (1 x 10(-1) M) of histamine when the nose was pretreated with saline, but was substantially stimulated by histamine in a dose-response fashion (1 x 10(-2) M) after pretreatment with HCI or H2SO4. Pretreatment with tripelennamine and HCl prevented the effect of histamine on the afferent EN activity; but after cimetidine and HCl pretreatment histamine still had a marked stimulant effect. In the case of single unit activities, histamine with HCl pretreatment had a long-lasting stimulatory effect (110.2 +/- 26.6 sec). It is concluded that the EN in guinea pigs include histamine-sensitive fibers whose sensitivity is mediated by H1 receptors and can respond to histamine only under abnormal conditions of the nasal mucosa.

Endotracheal cuff pressure as an index of airway smooth muscle activity: comparison with total lung resistance.

Pressure changes in the cuff of an endotracheal tube (Pcuff) were measured as an index of the tracheal smooth muscle activity and compared with total lung resistance (RL) in anesthetized, paralyzed and artificially ventilated dogs. After obtaining passive pressure-volume relationships of the cuff in situ, we activated the airway smooth muscle by electrical stimulation of the right vagus nerve, intravenous acetylcholine, and airway mechanical stimulation. The responses elicited by vagal stimulation and airway probing affected predominantly the tracheal smooth muscle, whereas acetylcholine administration caused homogeneous responses in Pcuff and RL, suggesting involvement of the smooth muscle of the entire airway. Pcuff cannot represent the whole airway smooth muscle activity, but it is more sensitive than RL for detecting vagally mediated smooth muscle responses. We conclude that the combination of Pcuff and RL may provide a better evaluation of smooth muscle response to various stimuli.

Asymmetry in reflex responses of nasal muscles in anesthetized guinea pigs.

Nasal reflexes elicited by mechanical or electrical stimulation of nasal afferents were studied in anesthetized guinea pigs. Probing the nasal cavity of one side evoked a greater activation of the contralateral than the ipsilateral nasal muscles and, occasionally, sneezing. Similarly, electrical stimulation of the ethmoidal nerve often caused sneezing, with a greater activation of the nasal muscles and a greater increase in resistance on the contralateral side. Asymmetrical activation of the nasal muscles in response to mechanical stimuli induces asymmetrical airflows, especially during sneezing, between the two sides of the nasal cavity. Most of the expired air is forcibly blown out through the ipsilateral nostril, thus improving the elimination of irritants from the nose.

The afferent activity of the superior laryngeal nerve, and respiratory reflexes specifically responding to intralaryngeal pressure changes in anesthetized Shiba goats.

This study was aimed at characterizing the superior laryngeal nerve (SLN) afferent activities under four different respiratory conditions, i.e., tracheostomy breathing (TB), upper airway breathing (UAB), tracheal occlusion (TO) and upper airway occlusion (UAO), and investigating respiratory changes in response to transmural pressures applied to the larynx in anesthetized Shiba goats. The activity recorded from the whole SLN increased at both inspiration and expiration during TB, UAB and TO, while an expiratory augmentation accompanied by an inspiratory inhibition was found during UAO. Based on recordings from 109 thin filament-preparations, 47 units were identified as 'drive' receptors, 31 as 'pressure' receptors (22 'positive' and 9 'negative' pressure receptors), and the rest 31 as 'non-modulated type' of receptors. The posterior cricoarytenoid (PCA) muscle activity showed a clear inspiratory modulation during UAB and was significantly enhanced by negative pressure applied to the isolated upper airway, where such an augmented activity was abolished by bilateral section of the SLN. No significant changes were found in the respiratory cycle during application of negative pressures to the larynx. The respiratory modulation of the SLN in Shiba goats was essentially identical to that reported for rabbits, rats and guinea pigs, but not in dogs. The reflex response of the upper airway muscles to the laryngeal pressure changes in Shiba goats were found to be less noticeable than in rabbits and dogs.

Bradykinin-induced airway contraction in two lines of guinea pigs with congenitally different airway sensitivity.

The airway responsiveness to bradykinin (0.1, 1 and 10 microg/kg, i.v.) was examined in two lines of guinea pigs, BHS (bronchial hypersensitive) and BHR (bronchial hyposensitive) lines, with different airway sensitivity to inhalation of acetylcholine (ACh)-aerosol. Normal Hartley strain guinea pigs were used as a control group. The airway contraction was measured by recording intratracheal pressure (P[IT]) and respiratory airflow (V) under the condition of artificial ventilation in anesthetized guinea pigs. The results show airway responsiveness to bradykinin in BHS guinea pigs to be significantly greater than in BHR and normal Hartley strain guinea pigs.

Nasal mechanoreceptors in guinea pigs.

The characteristics of nasal mechanoreceptors in the ethmoidal nerve (EN) of guinea pigs were clarified by electrophysiological identification of their responsiveness to transmural pressure, i.e., the inspiratory effort induced by tracheal occlusion and probing of the nasal cavity, vestibule or alae nasi of the nose. A total of 73 mechanoreceptors were recorded from 18 guinea pigs breathing through the nose or a tracheostomy with an isolated nasal airway. Six receptors (6/22) in nasal-breathing animals were stimulated by upper airway occlusion, and 18 receptors (18/22) in tracheostomy-breathing animals were stimulated by maintained negative pressure in the nose. Mechanoreceptors responding to probing to the nose were found in both experimental set-ups. The mean threshold of 'pressure'-responsive receptors to negative pressure was very high (-3.87 +/- 0.95 kPa). Most of the receptors were also examined for response to ammonia vapour or instillation of distilled water; only three 'touch'-responsive receptors could be stimulated by ammonia and/or distilled water. These results suggest low sensitivity to pressure changes and noxious chemical stimuli of mechanoreceptors in the EN of guinea pigs.

Cardiopulmonary responses to capsaicin instillation to the laryngeal lumen and their reflex mechanisms in rats.

Cardiorespiratory effects of capsaicin (CAPS) solution instilled into the larynx and the reflex mechanisms were investigated in rats with spontaneous breathing or under artificial ventilation. The first challenge with CAPS (100 micrograms/ml, 20 microliters) markedly inhibited spontaneous breathing due to a considerable prolongation of expiration time (TE) (1785% of control) in all rats. Circulatory changes such as hypertension (mean systolic blood pressure-210 mmHg) and bradycardia (10.5% decrease in heart rate) were also elicited by the 1st challenge with CAPS. These changes were largely reduced by the second challenge of CAPS; but not abolished in 3 of 5 rats by bilateral section of the superior laryngeal nerves (SLNs) and recurrent laryngeal nerves (RLNs). The bradycardia and hypertension after the CAPS-instillation were able to be elicited to the same extent in rats in the absence of apnea under artificial ventilation. The bradycardia was entirely abolished by pretreatment with atropine injection, while the hypertension was largely inhibited by phentolamine and propranolol. These results demonstrated that the laryngeal and/or pharyngeal noxious stimulus could induce marked cardiorespiratory reflexes, where the circulatory changes could be elicited in the absence of apnea, suggesting the presence of eliciting mechanisms of circulatory changes independent on the occurrence of apnea. In addition, it was also suggested that those cardiorespiratory responses were mediated by the afferents such as unmyelinated C-fiber endings and thin myelinated fibers and by the efferents, both the parasympathetic and sympathetic nervous systems.

Nasal receptors responding to cold and l-menthol airflow in the guinea pig.

The aim of this study was to demonstrate the presence of nasal 'cold' receptors, through recordings of action potentials from the ethmoidal nerve (EN), in guinea pigs and to characterize their responsiveness to l-menthol and capsaicin. Constant flows (400 ml/min) of room air (20 degrees C), warm air (45 degrees C), room air containing l-menthol, and cold air (-5 degrees C) were directed into the nasal cavity in the inspiratory direction via a nasopharyngeal catheter in the anesthetized guinea pigs breathing spontaneously through a tracheostomy. The ethmoidal afferent activity was increased by cold air, and to a greater extent by l-menthol but hardly by warm air. After topical anesthesia of the nasal cavity with 2% lidocaine, cold air and l-menthol no longer stimulated the EN. L-menthol noticeably stimulated the EN even after repeated capsaicin instillation into the nose, but these values were lower than those following the l-menthol stimulus before the 1st capsaicin treatment. These results suggest that the ethmoidal nerve in guinea pigs has cold-sensitive receptors which consist of both small myelinated fibers and C-fiber endings.

Nasal receptors responding to noxious chemical irritants.

This study was performed to investigate the chemoreception of trigeminal afferents in the nose. Single unit activity was recorded from the anterior ethmoidal nerve in the anesthetized guinea pig breathing through a tracheostomy during nasal instillation of capsaicin (0.3 mM), nicotine (6 mM) and ammonia (1.5 M) solutions or with distilled water. Out of 36 fibers recorded, nineteen were stimulated by capsaicin, six by nicotine and seventeen by ammonia. Among those fibers, two were stimulated by both capsaicin and nicotine, six by both capsaicin and ammonia and one nicotine-responsive fiber was also stimulated by ammonia. A large proportion of capsaicin- and nicotine-responsive fibers exhibited long lasting discharges (170.4 +/- 17.7 sec and 120.7 +/- 29.3 sec, respectively), and were not stimulated by the second application of the same substance. However, fibers responding to ammonia discharged for a shorter time (31.5 +/- 6.5 sec), indicating a rapid adaptation. These results indicate that the ethmoidal nerve possesses a well-developed responsiveness to noxious stimuli. The nociceptive component of this nerve may be related to the various cardiorespiratory responses that can be elicited from the nasal cavity and also to local axonal reflexes (neurogenic inflammation) due to the release of chemical mediators from C-fiber endings.

The respiratory activity of the superior laryngeal nerve in the rat.

The aim of this study was to characterize the laryngeal afferent activity of the rat. The animals were anesthetized and breathing spontaneously. Laryngeal afferent activity was recorded from both the whole superior laryngeal nerve (SLN) and from single fibers isolated from this nerve. An overall inspiratory augmenting activity was observed in the whole SLN during tracheostomy breathing, tracheal occlusion and upper airway breathing, but an expiratory augmenting activity was present during upper airway occlusion. The inspiratory modulated activity was abolished by bilateral section of the hypoglossal nerves but not the recurrent laryngeal nerves. A great number of receptors (46/80, 58%) were identified as 'drive' receptors, and others as 'pressure' (22/80, 28%) and 'irritant' type receptors (9/80, 11%). Nineteen pressure receptors were stimulated by positive transmural pressure, while only three stimulated by negative pressure. Nine drive receptors were also stimulated by positive pressure and inhibited by negative pressure. Such response to pressure was further evaluated by applying maintained pressures to the functionally isolated upper airway. These results are essentially consistent with findings obtained in the rabbit, but differ from those reported for the dog.