Respiratory Physiology & Neurobiology 152 (2006) 223–242

Vagal afferent nerves regulating the cough reflex
Brendan J. Canning a,∗ , Nanako Mori a , Stuart B. Mazzone b
a b

Johns Hopkins Asthma and Allergy Center, 5501 Hopkins Bayview Circle, Baltimore, MD 21224, USA Department of Neurobiology, Howard Florey Institute, University of Melbourne, Vic. 3010, Australia Accepted 1 March 2006

Abstract Coughing is initiated by activation of mechanically and chemically sensitive vagal afferent nerves innervating the airways. All afferent nerve subtypes innervating the airways can modulate the cough reflex. Rapidly adapting and slowly adapting stretch receptors (RARs and SARs, respectively) innervating the intrapulmonary airways and lung may enhance and facilitate coughing. Activation of intrapulmonary C-fibers has been shown to inhibit coughing in anesthetized animals. Extrapulmonary C-fibers and RARs can initiate coughing upon activation. C-fiber-dependent coughing is uniquely sensitive to anesthesia. Tracheal and bronchial C-fibers may also interact with other afferents to enhance coughing. Recent studies in anesthetized guinea pigs have identified a myelinated afferent nerve subtype that can be differentiated from intrapulmonary RARs and SARs and play an essential role in initiating cough. Whether these “cough receptors” are the guinea pig equivalent of the irritant receptors described in the extrapulmonary airways of other species is unclear. © 2006 Elsevier B.V. All rights reserved.
Keywords: Rapidly adapting receptor; Slowly adapting receptor; C-fiber; Capsaicin; Bradykinin

1. Introduction Coughing can be evoked in animals and in human subjects by inhalation of particulates, acids, irritant gases (e.g. sulfur dioxide), cigarette smoke, nicotine, capsaicin, bradykinin, prostanoids, hypo-

This paper is part of a special issue entitled “Cough and its Regulation”, guest-edited by John G. Widdicombe and Bradley J. Undem. ∗ Corresponding author. Tel.: +1 410 550 2156; fax: +1 410 550 2130. E-mail addresses: bjc@jhmi.edu (B.J. Canning), smazzone@hfi.unimelb.edu.au (S.B. Mazzone). 1569-9048/$ – see front matter © 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.resp.2006.03.001

and hypertonic solutions, and by mechanical probing of the tracheal, laryngeal and large bronchial mucosa (Widdicombe, 1954a; Gravenstein et al., 1954; Karlsson et al., 1988; Choudry et al., 1989; Hansson et al., 1994; Lalloo et al., 1995; Nishino et al., 1996; Karlsson and Fuller, 1999). Coughing is not so readily initiated by bronchospasm or by static changes in airway pressure (Chausow and Banner, 1983; Butler et al., 1995; House et al., 2004; Canning et al., 2004; El-Hashim and Amine, 2005). Regardless of the stimulus, coughing is attenuated by anesthesia (May and Widdicombe, 1954; Nishino et al., 1996; Tagaito et al., 1998; Canning et al., 2004) and completely abol-

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ished by vagotomy and/or vagal cooling (Widdicombe, 1954a; Klassen et al., 1951; Adams et al., 1987; Tatar et al., 1988; Canning et al., 2004). These and other observations indicate that the cough reflex is initiated by activation of chemically and mechanically sensitive vagal afferent nerves (Widdicombe, 1998). Afferent nerves innervating other viscera as well as somatosensory nerves innervating the chest wall, diaphragm and abdominal musculature play a less essential but important accessory role in regulating cough (Kondo et al., 1998; Morice et al., 2004). The mechanisms by which tussive stimuli activate the various afferent nerves regulating cough is reviewed in detail elsewhere in this issue (Kollarik and Undem, 2006). This review will describe the vagal afferent nerve subtypes innervating the airways and their proposed roles in regulating cough.

2. Classification of airway afferent nerve subtypes Airway afferent nerves are differentiated based on their physical and chemical sensitivity, adaptation to mechanical stimulation, neurochemistry, origin, myelination, conduction velocity, activity during tidal breathing, reflexes associated with their activation, and sites of termination in the airways, lungs and brain stem (Table 1; Fig. 1). The utility of the various approaches used to define subtypes of airway afferent nerves is limited largely by the lack of specificity of the characteristics displayed. For example, bronchopulmonary C-fibers are relatively selectively and directly activated by capsaicin and bradykinin (Coleridge and Coleridge, 1984; Riccio et al., 1996; Ho et al., 2001). But C-fiber activation, through parasympathetic or axonal reflexes, can initiate airways obstruction (e.g. mucus secretion, bronchospasm), a potent stimulus for rapidly adapting receptors (RARs; Coleridge and Coleridge, 1984; Jonzon et al., 1986; Karlsson et al., 1988; Mohammed et al., 1993; Bergren, 1997; Joad et al., 1997; Matsumoto et al., 1997; Canning et al., 2004). Similarly, rapid adaptation to sustained lung inflation is a defining characteristic of RARs, but RARs may adapt poorly to lung deflation and bronchospasm (Bergren, 1997; Ho et al., 2001; Canning et al., 2004). Interventions and stimuli thought to have selective effects are often nonselective. For example, sulfur dioxide has been used as a method to inactivate SARs, resulting

in a suppression of the cough reflex and leading to the hypothesis that ongoing SAR activity is critical to the initiation of cough (Hanacek et al., 1984). But sulfur dioxide activates bronchopulmonary C-fibers (Atzori et al., 1992; Wang et al., 1996), and pulmonary C-fiber activation has also been associated with a suppression of cough (Tatar et al., 1988, 1994). Species differences are also apparent. The axon reflex and its associated mechanical effects (caused by vascular engorgement, plasma exudation, bronchospasm and mucus secretion) is well defined in the airways of rats and guinea pigs, but has not been established in the airways of other species, including humans (Barnes, 2001). This may be due to the paucity of neuropeptide (substance P, neurokinin A (NKA) and calcitonin gene-related peptide (CGRP)) expression by airway C-fibers in species other than guinea pigs and rats (Lundberg et al., 1984; Baluk et al., 1992; Riccio et al., 1996; Lamb and Sparrow, 2002). The distribution of afferent nerve subtypes within the airway also differs amongst species (Schelegle and Green, 2001). Another complicating factor relating to afferent nerve subtype differentiation is the considerable overlap in portions of the airways and lungs innervated by the identified subtypes, along with similar reflex effects initiated upon their activation in some instances. Finally, search paradigms and even the definitions for subtypes of airway afferent nerves differ from laboratory to laboratory (Widdicombe, 1954b; Paintal, 1973; Coleridge and Coleridge, 1984; Bergren and Peterson, 1993; Ho et al., 2001; Canning et al., 2004). All of these limitations to current approaches used to define afferent nerve subtypes, and the spectrum of responses evoked by specific stimuli (Widdicombe, 1954b; Miserocchi and Sant’Ambrogio, 1974; Pack and DeLaney, 1983; Coleridge and Coleridge, 1984; Yu, 2000), should be considered when evaluating both the published literature and new experimental data and their relevance to the study of cough.

3. Properties of airway afferent nerve subtypes and their potential role in regulating cough 3.1. Rapidly adapting receptors RARs terminate in the intrapulmonary airways of all species studied and in the extrapulmonary airways of dogs and cats. Anatomically, the structure of RAR

suggestive of small. e Stretch refers to a mechanical stretch in either the longitudinal or circumferential axis of the airway. 2006)..g. Canning et al. bradykinin or capsaicin can be markedly reduced or abolished when the resulting airway smooth muscle contraction and/or bronchospasm is prevented. 1996). myelinated axons) (Tables 1 and 2.. or to a distending pressure (20–100 cmH2 O). a Cough receptors have been described in detail elsewhere (Canning et al.g. additional subtypes that have been described (e.. adenosine. g Responsiveness to ATP has only been studied in vitro in guinea pigs (Undem et al. Canning et al. methacholine. their responsiveness to lung collapse and/or lung deflation. See text for further details. Undem et al. 2004). 2004. The mechanisms by which these stimuli activate afferent nerves have been reviewed elsewhere in this issue (Kollarik and Undem. Canning et al. accessibility via the bronchial or pulmonary circulation) and stimulants (e. and their conduction velocity (4–18 m/s. terminations in the airway wall is poorly described (Widdicombe. b C-fiber subtypes have been described elsewhere (Coleridge and Coleridge. NA: not assessed.B. 2004) and cats (Kubin and Davies. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 Table 1 Characteristics of vagal afferent nerve subtypes innervating the larynx.. f Activation of RARs by histamine.. but are readily distinguished from intrapulmonary RARs in most species.. For clarity. c Ganglionic origin has been determined only in guinea pigs (Riccio et al.g.. The sustained activation of RARs produced by dynamic lung inflation. 2). 2004). trachea. so only the most commonly reported attributes are listed. 1984. Physiologically. bronchospasm or lung collapse indicates that adaptation of RARs is not attributable . reflex effects. Fig. 1996. 2003).g. 1). All subtypes are responsive to punctate mechanical stimulation but perhaps with differing sensitivities. 2001. RARs are also distinguished from other sub- types by their responsiveness to alterations in dynamic lung compliance (and thus their sensitivity to bronchospasm). capsaicinsensitive Adelta fibers in guinea pigs) have been omitted. but C-fibers may also be subdivided into bronchial (neural crest-like) and pulmonary (placodal-like) subtypes. RAR terminals are dynamic receptors that respond to changes in airway mechanics (e. The embryologic distinction proposed by Undem et al. These may correspond to the subset of extrapulmonary RARs (“irritant” receptors) described in other species. 1995). bronchi and lungs Anatomical properties SARs RARs Cough receptorsa C-fibersb Neural crestb Ganglionic originc Intrapulmonary terminations Extrapulmonary terminations Neuropeptide synthesisd Physiological Properties Conduction Velocity (m/s) Activity during tidal breathing (impulse/s) Lung inflation/stretche Adaptation to lung inflation Lung deflation Carbon dioxide Acid Hypertonic saline Pulmonary embolism Pulmonary edema/congestion Bronchospasmf Pharmacological properties Bradykinin ATP 5-HT Capsaicin Nodose Yes Yesd No 15–32 10–50 Activated Slow No effectd Inhibited NA NA Sensitized Variable Activated No effect Activatedg No effect No effect Nodose Yes Yes No 14–23 0–20d Activated Rapid Activated No effect NA Activated Activated Activated Activatedf Activatedf Activatedg Activated Activatedf Nodose Few Yes No 4–6 NA No effect No response No effect NA Activated No effect NA NA No effect No effect No effect No effect No effect jugular Yes Yes Yes ∼1 <2 No effect No response No effect NA Activated Activated NA NA No effect Activated No effect No effect Activated 225 Placodalb nodose Yes Few Some ∼1 <2 Activated Slow No effect Activated NA NA Activated Activated No effect Activated Activatedg Activated Activated Wide variation in responsiveness and physiologic characteristics has been noted in all subtypes. Characteristics that distinguish the various subtypes are indicated in italics. “intermediate” receptors in cats. They are functionally differentiated from other airway afferent nerves by their rapid (1–2 s) adaptation to sustained lung inflations (Fig. Additional characteristics (e. diameter. interstitial and intraluminal pressures). length. sulfur dioxide) have been reported.J. d Marked species differences have been noted. 2004) and in one study in dogs (Pelleg and Hurt. Undem et al. 2004. is presented here.

a In spontaneously breathing dogs (Pisarri et al.. Bergren and Sampson..7 7. the Table 2 Effect of decreased lung compliance (Cdyn) on airway afferent nerve activity Afferent nerve subtypes Species Action potentials (impulse/s) Baseline Low threshold SARs (n = 12) High threshold SARs (n = 12) SARs (n = 11) RARs (n = 31)a RARs (n = 17) RARs (n = 26) RARs (n = 27) RARs (n = 14) Bronchial C-fibers (n = 23) Pulmonary C-fibers (n = 4) Cat Cat Rabbit Dog Rabbit Rabbit Cat Dog Rabbit Rabbit 54 19 28 1. Canning et al.. Cdyn was decreased 10–50% in cats (Yu et al.2 ± 0. * Denotes a statistically significant increase in afferent nerve activity when Cdyn was decreased. 2003) and dogs (Jonzon et al. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 Fig.0 2. RARs discharge during lung inflation and sometimes during the deflation phase of a respiratory cycle and become more active as the rate and volume of lung inflation (and/or deflation) increases (Pack and DeLaney.. open chest rats. 2001). 1983. Jonzon et al. 2001). In artificially ventilated.8* 1.. Recordings are representative of 17–55 experiments on fibers of each subtype (figure modified with permission from Ho et al.9 ± 0. 1984.2 1 ± 0.1 0.. Ma et al. (B) rapidly adapting receptors.3 0.. 1974.2 1..1 2.. Responsiveness to capsaicin and sustained lung inflation highlight the primary differences amongst the three major subtypes of intrapulmonary vagal afferent nerves (see text for further details).3 0. 1991). Ho et al. 2001.8 9. 1990). Ho et al. Pack and DeLaney.. 2004).J. 1987. 1986) by altering end-expiratory pressures.2 ± ± ± ± ± ± ± ± 4 2 3** 0. 2001. 1954b. Yu et al. 1982. Canning et al. rabbits (Yu et al. 1.6 3 7.9 Decreased Cdyn 50 18 22 3. Coleridge and Coleridge. . However.6* 0.1 ± 0.6 ± ± ± ± ± ± ± ± 3 2 3 0. 1983. RARs are relatively insensitive to “direct” chemical stimuli. 2001. ** Denotes a statistically significant decrease in afferent nerve activity when Cdyn was decreased. Cdyn was decreased 25–50% by restricting chest wall movements. Sant’Ambrogio and Widdicombe.226 B. 2003. Representative recordings illustrating the basal activity and responsiveness of (A) C-fibers. to an electrophysiological adaptation (Widdicombe.3 Experiments were carried out in either open or closed chest preparations.. Armstrong and Luck.1 1.5* 0.8* 1.5* 1. Afferent nerve subtypes were identified based on their conduction velocity and/or their adaptation to sustained lung inflation. 1986.1* 0. and (C) slowly adapting receptors in anesthetized. 1987. open chest preparations.3 0. 1987.2 0.

1996... El-Hashim and Amine. substance P and bradykinin augments and/or mimics the mechanical consequences of lung inflation and deflation and thus increases RAR activity (Bergren and Sampson. Canning et al. intrapulmonary RARs are unlikely to regulate cough..g. Widdicombe. 1993. inspiratory and expiratory efforts against a closed glottis) are mostly ineffective at evoking cough (Chausow and Banner..J. 2003). 2002). 1997. Shinagawa et al. 1954b. Butler et al. 1997. 2001). Fujimura et al. Note that unlike the adaptation to sustained lung inflation. 1983. mucus secretion. histamine. thromboxane. Lungs were inflated (A) with constant flow (20 L/min) to various volumes.B. 1994. The RAR discharge and reflexes initiated by these stimuli can be markedly inhibited or abolished by pharmacologically preventing their end organ effects (e. 1994. However. Widdicombe. 1982. 1987. Joad et al.. Individual responses (panel A) and mean data from 17 RARs (adaptation index: >90) illustrating the dynamic responsiveness of RARs in dogs.. 1988. Canning et al. Bonham et al. RARs continue to respond incrementally to dynamic lung inflation. 2... arguing for an indirect mechanism of action.. 2003. methacholine. neurokinins. Sant’Ambrogio et al. Canning. . 2003). RARs may also adapt poorly to (C) bronchospasm (evoked in guinea pigs) and (D) lung deflation (evoked in rats) (figure is modified from Pack and DeLaney. 1986. Tatar et al. bronchospasm). Morikawa et al.. Many stimuli that activate intrapulmonary RARs (e. or (B) to a constant volume (three times basal tidal volume) at various flows. Barnes et al. RARs are dynamic receptors. Mohammed et al.. 1992. Joos et al. Canning et al. 2005). 1997. 2001. 1984. RARs respond to stimuli that evoke cough and fulfil many of the accepted criteria for mediating this defen- sive reflex (Widdicombe. 1988. Bergren. 1997. 2004.. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 227 Fig. 2000. Bergren.g.. Widdicombe. Further evidence for their role in cough comes from vagal cooling studies. 1998.. Jonzon et al. capsaicin. leukotriene C4 (LTC4 ). 1983. bronchospasm and obstruction resulting from mucus secretion or edema evoked by substances such as histamine.. 1997. 1995. Ho et al.. which blocks cough at temperatures that abolish activity in RARs (and SARs) while preserving C-fiber activity (Tatar et al.

Schelegle and Green. 2004). 2006). In cats. In rats. Many SARs have multiple receptive fields (Yu and Zhang. 1. Given their profound influence over respiratory pattern and their activity throughout nearly all phases of the respiratory cycle. SAR activity increases during inspiration and peaks just prior to the initiation of expiration (Miserocchi and Sant’Ambrogio. Ho et . 1984. SAR activation results in central inhibition of respiration and inhibition of cholinergic drive to the airways.. rabbits and rats have few SARs in their extrapulmonary airways (Schelegle and Green. SAR dendritic arbors were associated with the bronchiolar smooth muscle. 3. 1974). In dogs. 1984. SARs innervate the trachealis and are activated by tracheal smooth muscle contraction (Krauhs. 2001). Responsiveness to static lung inflation at various transpulmonary pressures differentiates subtypes of intrapulmonary slowly adapting receptors (SARs). 2004). 2002. SAR terminals were not associated with neuroepithelial bodies (Yu et al. 2001). however. 3). 1988) and both capsaicin and bradykinin induce coughing in conscious animals but do not activate SARs (Coleridge and Coleridge... As mentioned above. Canning et al.E. and rats.2.b) provided definitive evidence that extrapulmonary RARs regulate coughing in cats and dogs. Table 1).. Sant’Ambrogio et al. 1988). Schelegle and Green. Fig. of 8–39 experiments (figure is modified from Miserocchi and Sant’Ambrogio.g. or perhaps a different afferent nerve subtype regulates coughing in this species (see below). Comparable subtypes of SARs innervating the extrapulmonary airways were also described in this study carried out in dogs. Yu and Zhang. Schelegle and Green. but not uniformly. Richardson et al. Ho et al. 2001). 2004). and in most species by their modest adaptation to sustained lung inflation (Fig. Occasionally. The majority of SARs studied produced incrementally more action potentials as static transpulmonary pressure increased.M. 2004. SARs may be differentially distributed in the airways of commonly studied mammalian species (Schelegle and Green. SARs almost certainly influence coughing. which terminates inspiration and initiates expiration when the lungs are adequately inflated (Schelegle and Green. 1984. SAR terminations were found primarily in the peripheral airways (associated with alveoli or bronchioles). have described the structure of electrophysiologically identified SARs innervating the intrapulmonary airways and lungs of rabbits and rats (Yu et al. Subtypes of SARs have been proposed (Miserocchi and Sant’Ambrogio. In dogs.J. 2001. 3. The sensory terminals of SARs have a complex structure with varied sites of termination and extensive branching.. SARs also differ from RARs with respect to the reflexes they precipitate. 2004. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 RARs throughout the airways may. laryngeal or mainstem bronchial mucosa (Canning et al. House et al.. 1988). But RARs as described by Widdicombe have not been described in the extrapulmonary airways of guinea pigs even though this species coughs readily to punctate mechanical stimulation or acid applied topically to the tracheal. SARs can be differentiated from RARs in some species based on action potential conduction velocity. Widdicombe (1954a. 1974. SARs are thought to regulate the Hering-Breuer reflex. 2001) (Fig. Perhaps the definition of RARs could be expanded to include the afferent nerves mediating cough in guinea pigs (e. But SAR activity does not increase prior to or during ammonia-induced coughing in rabbits (Matsumoto. “irritant” or “cough” receptors). guinea pigs. 2001). Yu et al. Data represents the mean ± S. 2003. leading to decreased phrenic nerve activity and decreased airway smooth muscle tone (due to a withdrawal of cholinergic nerve activity. Sant’Ambrogio et al. SARs may also be localized to extrapulmonary airways (Miserocchi and Sant’Ambrogio. 1974.228 B. act synergistically with other afferent nerve subtypes to induce coughing (Canning. Slowly adapting stretch receptors SARs are responsive to the mechanical forces associated with breathing. cats.. 2004). few SARs but many RAR-like receptors and C-fibers can be found in the extrapulmonary airways. guinea pigs. 2001).. 1974.

. Considering all of the available data on SARs and cough. 2003). Enhanced baseline SAR activity produced by the loop diuretic furosemide has... Kwong and Lee. Lee and Pisarri. 2002. .. 1999. 2002).. 3. 1999. 2001. and thus embryologic. Baluk et al. 2002. 1993). Such data along with patch clamp recordings from the cell bodies of airway afferent nerves indicate that bradykinin and capsaicin (along with acid. 1984. or in rabbits. a species that has only a weak cough response (Larson et al. been associated with the reported antitussive effects of this agent (Sudo et al. may either enhance or inhibit coughing (Schelegle and Green. Very little is known about the distribution of NEBs and PNECs in the airways of species known to cough and in cats at least. 1992.. Kollarik et al. 1984. 2005. Recent studies of pulmonary neuroendocrine cells (PNECs) and neuroepithelial bodies (NEBs) suggest that TRPV1 expressing. 2006). C-fibers are further distinguished from RARs by the observation that bradykinin and capsaicin-evoked activation of their endings in the airways is enhanced by PGE2. Most studies of NEBs have been carried out in rats. 2001.... which increases baseline SAR activity. Conversely.. Mitchell et al. 2003. it seems reasonable to conclude that SAR activity neither precludes coughing nor is sufficient for evoking cough.. Weichselbaum et al. 1992. 1992. Lee and Pisarri. in press) directly activate bronchopulmonary C-fibers (Fig. applying positive airway pressures.. Lee et al. ganglionic. Modeling activation of brainstem second order neurons (termed pump cells) of the SAR reflex pathway predicts facilitation of coughing (Shannon et al. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 229 al...... 2004). NEBs are reported to largely disappear from the large airways soon after birth (van Lommel and Lauweryns. Mazzone and Canning. TRPV1 may also be used to localize the peripheral terminals of C-fibers (Myers et al. 1996.. Dey et al. mice and hamsters. C-fibers Bronchopulmonary C-fibers are distinguished from RARs and SARs by their conduction velocity. which would decrease basal SAR activity. Riccio et al. 2002. Table 1). 2005. olvanil and resiniferotoxin.. Groneberg et al. 2001. 1984.J. 2001). Pan et al. 2001.. Riccio et al. Studies of CNS processing have argued for both inhibitory and facilitating roles for SARs in coughing. 2004. Hypercapnea. Hunter and Undem.. Canning et al. responsiveness to chemical and mechanical stimuli and in guinea pigs. Coleridge and Coleridge. 1984. 1. 2002). 1996. 1996. Van Genechten et al. 2004. 2004. 2003. origin (Coleridge and Coleridge. CGRP-containing spinal afferents may innervate these epithelial cells and structures (Brouns et al.. increases expiratory efforts during cough in animals (Hanacek and Korpas. 2005.. Notably. It is at present unclear whether NEBs or PNECs play any role in regulating cough.3. 2006). 2005). Gu and Lee. however. Bolser et al.B.. Undem et al.. 1990. 1982) but not humans (Nishino et al. Lamb and Sparrow. Much about the role of SARs in regulating cough and other defensive reflexes remains poorly defined. 1974. Bergren.. 1984. however.. Watanabe et al. adrenaline and adenosine. Airway C-fibers can be subdivided into bronchial and pulmonary C-fibers.. But an excitatory role of pump cells in cough is difficult to reconcile with studies showing that SARs (via pump cells) inhibit RARmediated reflex pathways (Ezure and Tanaka. 2004). Chuaychoo et al.. Undem et al.. bronchodilators that would largely oppose the end organ effects associated with these stimuli and the resulting C-fiber activation (Ho et al. 2005).. 1997. Myers et al. 2005).. 1998. Widdicombe and Singh. This unique neurochemical property of bronchopulmonary C-fibers has been exploited to describe the distribution and peripheral terminals of these unmyelinated afferent nerve endings in the airway epithelium as well as in other effector structures within the airway wall (Lundberg et al.. Undem et al.. 2004. Ho et al. to distinguish TRPV1 expressing C-fibers from myelinated TRPV1 expressing vagal afferent nerves or capsaicin sensitive spinal afferents innervating the airways (Kummer et al. 2000). Canning et al. 2000. Hunter and Undem. Also arguing for a facilitating role of SARs in cough would be experiments performed on rabbits in which inhaled sulfur dioxide blocks SAR activity and coincidentally attenuates coughing (Sant’Ambrogio et al. their relative insensitivity to mechanical stimulation and lung inflation and their responsiveness to bradykinin and capsaicin (Armstrong and Luck.. Oh et al. C-fibers (particularly in rats and guinea pigs) may synthesize neuropeptides that can be localized to their central and peripheral terminals (Lundberg et al. Hanacek et al. species that lack a cough reflex. 1984).. 1989). 2002. 2005). Baluk et al.. It is at present impossible. 1990. a distinction based on sites of termination. 2000). Riccio et al. Dey et al.

By contrast.. 1996. 1993. Canning et al. Ho et al. C-fiber endings are polymodal. 1993... C-fibers are thus generally quiescent throughout the respiratory cycle but are activated by chemical stimuli such as capsaicin. 2002). pharmacological studies that take advantage of the unique expression of tachykinins by airway C-fibers have shown that bradykinin. C-fiber subtypes in guinea pigs have been identified based on their ganglionic origin and sites of termination in the airways (Undem et al. Karlsson et al...J. Ho et al. histamine appears to be without effect on bronchopulmonary C-fibers. bradykinin. 2005) (Fig. Karlsson. Capsaicin.. Barnes. the stimuli that activate intrapulmonary RARs and SARs but do not readily activate C-fibers or evoke coughing.. responding to both chemical and mechanical stimulation (Fig. particularly in anesthetized animals. Substantial evidence suggests that C-fiber activation.. C-fibers with cell bodies in the nodose ganglia terminate almost exclusively in the intrapulmonary airways. Systemic administration of C-fiber stimulants ... bradykinin. Lee et al. Finally. Forsberg and Karlsson. Canning et al. 5-HT or adenosine but are activated by capsaicin. Palecek et al.. 2004. These C-fibers are not activated by ATP. 2005)... 2001). Capsaicin desensitization abolishes citric acid induced coughing in guinea pigs..... Reflex responses evoked by C-fiber activation include increased airway parasympathetic nerve activity and the pulmonary chemoreflex. 2001. 1993. Widdicombe. 1996. Lin and Lee. are activated by ATP. In guinea pigs. 1988... are activated by capsaicin and bradykinin but unlike jugular C-fibers. 1986). electrical or chemical (acid) stimulation of the laryngeal. Jia et al. Choudry et al. In mice. Bolser et al. 2002). 2004. 2002. including antagonists with little or no capacity to cross the blood–brain barrier (Bolser et al. 1984). 1984. even though cough can be induced in these animals by mechanical. hypertonic saline and acid. Nevertheless. Lee and Pisarri. Kollarik and Undem. sulfur dioxide and citric acid evoke cough in conscious animals and in humans (Coleridge and Coleridge. 1996. Undem et al. but has no effect on cough evoked by mechanical probing of the airway mucosa in these same animals (Forsberg and Karlsson. 2001. while bronchial C-fibers are activated by histamine (Coleridge and Coleridge. However. cats.. ATP activates all C-fibers whereas capsaicin and bradykinin activate only a subset of the identified bronchopulmonary Cfibers (Kollarik et al. 2001. methacholine) and maximal inspiratory or expiratory efforts against a closed glottis. 1989. hypertonic saline and sulfur dioxide (Coleridge and Coleridge. characterized by apnea (followed by rapid shallow breathing). 1994. LTC4. Mohammed et al. citric acid. 1989. substance P.. C-fiber stimulation has consistently failed to evoke coughing in anesthetized animals. tracheal or bronchial mucosa (Tatar et al. C-fiber activation in guinea pigs and rats has also been associated with the axon reflex (Lee and Pisarri. 1995. 2001. 1997. Lee and Pisarri. 1991.230 B. C-fibers arising from the jugular ganglia in guinea pigs innervate both intrapulmonary and extrapulmonary airways and almost uniformly express the neuropeptides substance P and CGRP. 1996. 2001). bradykinin. 2004). capsazepine and iodo-resiniferotoxin inhibit capsaicin and acid evoked coughing in awake animals (Bolser et al.. Mazzone et al. 2001). Lalloo et al.. 2004). also argue for a role of C-fibers in cough.g. including bronchoconstrictors (e. Tables 1 and 2). 4). 2002.. adenosine and 5-HT (Undem et al. Lee and Pisarri. Chapman et al. Other openers of TRPV1 including anandamide and resiniferotoxin initiate coughing in animals and human subjects (Laude et al. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 pulmonary C-fibers in dogs may be unresponsive to histamine. 2002. 2004).. Deep et al. Myers et al. 2003). Wang et al. Ho et al. Trevisani et al. 2004).. 1997.. histamine. expressed almost exclusively by C-fibers (Riccio et al. capsaicin and mechanical stimulation-induced cough in dogs. “bronchial” C-fiber activation in particular. 1998. C-fiber thresholds for mechanical activation are substantially higher than that of RARs and SARs (Coleridge and Coleridge. These C-fibers do not consistently express substance P under normal conditions. Mazzone et al.. Lin and Lee. in press). 2001. 2004. 2001. Riccio et al. initiates coughing in conscious animals and human subjects. 1984. Canning et al. guinea pigs and pigs is attenuated or abolished by neurokinin receptor antagonists. By exclusion. citric acid. 1984. 1986... 1984. 2001. Mazzone et al. Stimulants of bronchial C-fibers such as capsaicin... while the TRPV1 blockers and antagonists ruthenium red. 1. 2002. acid and bradykinin work entirely or partially through activation of the ion channel TRPV1.. There is also considerable evidence to suggest that activation of airway C-fibers does not evoke cough and may actually inhibit cough. bradycardia and hypotension (Coleridge and Coleridge. 2002. Undem et al. Advenier et al.

anesthesia must selectively inhibit cough-related neural pathways. 1988.. Canning et al. 2001. 5. 1988. 1996. Pisarri et al... 1993. 2001). The fact that vagal cooling to temperatures that preserve C-fiber-dependent reflexes yet abolish cough is further evidence against a role for Cfibers in cough (Widdicombe. 1997.. 1984. Tatar et al. Davis et al. 1993). Bergren. Rather. Tatar et al. (C) Mean data from 12 conscious and five anesthetized guinea pigs (this figure was modified with permission from Canning et al.J. That capsaicin-evoked cough can be consciously suppressed in human subjects argues for an important role of perception on C-fiber-dependent cough (Hutchings et al. Canning et al. 1982. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 231 Fig. or may accentuate the inhibitory effects of C-fiber activation on cough. Alternatively.. 5). It is unclear what accounts for the differences in cough responsiveness between awake and anesthetized animals and human subjects (Fig. 1994). Bergren.. 4.B. It seems unlikely that a Fig. potentiates coughing in guinea pigs (Karlsson et al.. . 1981. Coleridge and Coleridge. Acid but not capsaicin applied topically to the tracheal mucosa of anesthetized guinea pigs evokes coughing. general anesthesia may interfere with the conscious perception of airway irritation and the resulting urge to cough. Tatar et al. 1994). which carry primarily C-fibers to the airways. 1991.. Canning et al... Bradykinin (10 mg/mL inhaled) evokes coughing in (A) conscious guinea pigs but (B) evokes no coughing following anesthesia with urethane. Severing the superior laryngeal nerves. 2004). 2004). 1998.. 2004). Anesthesia does not prevent C-fiber activation and/or C-fiber-mediated reflex effects entirely (Roberts et al. Traces are representative of more than 10 separate experiments (the figure is modified with permission from Canning et al. can inhibit cough evoked by mechanical stimulation (Widdicombe.. Higher concentrations of acid still evoke cough but also initiate effects on respiration comparable to that evoked by capsaicin. Note that capsaicin evokes a marked slowing of respiration and apnea followed by gasping and eventual restoration of normal respiratory rate. 1998.

These myelinated afferents are activated by acid. Rather. SARs and C-fibers are rigidly defined. conscious suppression of cough would be effective against an aspiration-induced cough (which likely involves activation of afferents in addition to Cfibers). “irritant” or “cough” receptors) that are mostly insensitive to capsaicin that can initiate cough upon activation in both awake and anesthetized animals. 1984. Ho et al. acetone vapor). trachea and mainstem bronchi (Riccio et al. It has also been argued that many of the putatively selective C-fiber stimulants may also activate RARs. This has been documented to some extent. 1997. acid and other irritating chemicals. 1989). 3. bradykinin. 1997. The inability of C-fiber activation to initiate cough following anesthesia also argues against this peripheral interaction. Such a population of afferents has not been described in other species (although they certainly may exist). these afferents do not rapidly adapt to mechanical stimulation (McAlexander et al. 1996).J. Such experiments await a better understanding of CNS control of cough.. rendering these data somewhat suspect.000-fold) its potency at any known neurokinin receptor. acid and bradykinin on C-fibers are very consistent and robust while their effects on RARs has been highly variable and subtle (Coleridge and Coleridge. the peripherally released neuropeptides either sensitize or directly activate RARs and/or other airway mechanoreceptors to initiate or facilitate coughing (Canning. What differs most is the way cough is studied in different species (mostly in conscious humans and guinea pigs. and their importance is obscured in population studies in which RARs. Canning et al. The peripheral pathways of these two types of cough have been established. punctate mechanical stimulation and perhaps by volatile gases (e. there are populations of myelinated vagal afferents (e. But. as anesthesia does not prevent the axon reflex or mechanically evoked cough in guinea pigs. Bergren. may account for the ability of C-fiber activation to initiate cough. SO2 . bradykinin and acid consistently and reliably initiate coughing in conscious individuals of all species studied and these stimuli are generally selective for C-fiber activation. 2002). 2005).. A counterargument has been that the C-fiber-dependent axon reflex. Morikawa et al. Conversely.. Cfiber-independent cough is less sensitive to anesthesia and initiated by mechanical stimulation and acid.. 2005). Two types of species that cough or two types of cough? Drawing the wholly dissatisfying conclusion that C-fibers regulate coughing in some species but not in others is tempting but seems unnecessary. C-fiber-dependent cough is sensitive to general anesthesia and initiated by capsaicin. but the effects of stimuli such as capsaicin. Only one group seems to have been able to evoke coughing with inhaled substance P (Kohrogi et al. Essentially every known stimulant of airway C-fibers evokes coughing in this species and nearly every drug that either selectively prevents C-fiber activation or prevents the actions or release of the neuropeptides uniquely expressed by C-fibers also prevents coughing. 1999) and do not seem capable of initiating cough upon activation in anesthetized guinea pigs (Canning et al. In guinea pigs.g. Future progress towards testing this hypothesis will likely rely on studies of CNS processing of the cough reflex and perhaps functional imaging of the brain during cough evoked by activation of these parallel pathways.g. Perhaps a subset of RARs that regulate cough are activated by stimuli such as capsaicin and bradykinin. . Mohammed et al.. In this scenario. so well defined in guinea pigs and seemingly nonexistent in all other species that possess a cough reflex. 2001). even though citric acid induces coughing without inducing much airways obstruction (El-Hashim and Amine. we think there are two types of cough — C-fiber-dependent and C-fiberindependent coughs.. this now seems unlikely.232 B. Many studies of citric acid or capsaicin-evoked cough are carried out following pretreatment with the -adrenoceptor agonist terbutaline to limit the airways obstruction evoked.4. Inhaled tachykinins have been shown in most studies to be ineffective at evoking cough (Advenier et al. RARs. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 comparable.. 1993. for example.1 fM in an aerosol) far exceeds (by 10. but the potency of the agonist in these studies (0. The evidence favoring a role for C-fibers in cough in conscious guinea pigs is overwhelming. 2004). mostly in anesthetized cats and dogs). there is a subpopulation of myelinated (A ) capsaicin sensitive afferent nerves that innervate the larynx. Stimuli such as capsaicin. but in any event.. The mechanisms of cough probably do not differ significantly amongst species.

Immunohistochemical analyses reveal that the putative cough receptors. with complex dendritic arbors arranged in a circumferentially around the airway. 5-HT. Winner et al.. Pedersen et al. Fujimura et al. Pedersen et al.. Selective nerve cuts and an analysis of the stimuli that evoke coughing and afferent nerve activation in guinea pigs indicate that the tracheal. 2000. 2003). 2004. the cough receptors are unresponsive to several spasmogens and autacoids that induce airway smooth muscle contraction including histamine. These endings (∼150–200 along the entire length of the guinea pig trachea) are located in the mucosa. the capsaicin receptor (Riccio et al. between the smooth muscle and the epithelium (which may account for their insensitivity to smooth muscle contraction). being activated by punctate mechanical stimuli. Canning et al. Such distending stimuli do not evoke coughing in guinea pigs. 2002). 2004. But several characteristics distinguish the cough receptors from classically defined RARs. acid and the potassium channel blocker 4-aminopyridine (Riccio et al... express an isozyme of the sodium pump containing the 3 subunit. they are unresponsive to capsaicin. 1996. Canning et al. SARs and C-fibers plays an essential role in regulating the cough reflex (Canning et al. mechanical or electrical stimulation of the tracheal and laryngeal mucosa in anesthetized guinea pigs while having no effect on C-fiber-dependent reflexes evoked from the trachea. LTC4 . but with little regard for any accepted nomenclature for defining afferent nerve subtypes in the airways. renal mechanoreceptors and intrapulmonary SARs (Chapleau et al. 1983.. whereas intrapulmonary RARs in guinea pigs conduct action potentials at a much faster rate. 1993.. with terminals located primarily ipsilateral to their vagal origin. Canning et al. 2003. 1996. 2000. 1996... McAlexander and Undem. Their myelination.. Canning et al. Shinagawa et al. 2005). 2004). NKA. All of these stimuli activate RARs and yet each is generally ineffective at inducing cough (Chausow and Banner. 2000.. The unique expression of this isozyme is of interest.B.. Recent studies in anesthetized guinea pigs indicate that an airway afferent nerve subtype distinct from RARs. The cough receptors may also be distinguished from RARs based on conduction velocity. 2002) (Fig.. but not C-fibers. upwards of 15 m/s (Bergren and Sampson. Furosemide may also inhibit cough receptor activation (Karlsson and . Canning et al.. 2006). Joos et al. Unlike RARs. The left and right vagus nerve contribute approximately equal numbers of these receptors. These afferent nerves are polymodal. 6). / Respiratory Physiology & Neurobiology 152 (2006) 223–242 233 3.. particularly in the clinical literature. given its association with mechanoreceptors and not Cfibers in the somatic nervous system (Dobretsov et al.. 2003).. substance P. The terminal adhesions of the cough receptors and the characteristic orientation within the tissue indicate that they have attached to components in the extracellular matrix comprising the basement membrane. Because they are myelinated and adapt rapidly to a punctate mechanical stimulation (McAlexander et al. This inhibitory effect of ouabain on the cough receptors contrasts sharply with the ability of this compound to greatly enhance the excitability of baroreceptors. We have identified the receptive fields of cough receptors in the guinea pig tracheal and bronchial mucosa (Mazzone and Canning. methacholine. 2001. Widdicombe. Kopp et al. 2003. 1998. Cough receptors. 1994. The sodium pump inhibitor ouabain reduces or abolishes coughing evoked by topically applied citric acid. However. Canning et al. 2006) (Fig.. 1996. 1987. it is tempting to conclude that the cough receptors are merely RARs that innervate the extrapulmonary airways.. Riccio et al. 2004). even pressure changes exceeding −100 to +100 cmH2 O (Canning et al. Myers et al. Widdicombe. 2006). 2004).. 1992. Barnes et al. 2005). The cough receptors assume a stereotypical position in the airway wall. Cough receptors innervating the larynx. ATP and adenosine (Riccio et al. 1998. In vitro electrophysiological recordings confirm the potent and selective inhibitory effects of ouabain on the putative cough receptor. Evidence for a “cough receptor” The term “cough receptor” has been used routinely in the literature... 1982. El-Hashim and Amine. trachea and bronchus of guinea pigs conduct action potentials at ∼5 m/s.. laryngeal and bronchial afferent nerves primarily responsible for regulating the cough reflex arise from the nodose ganglia.J. conduction velocity and insensitivity to capsaicin and bradykinin clearly differentiate the putative cough receptors from bronchopulmonary C-fibers. Matsumoto et al. 1999)...5. bradykinin or hypertonic saline and do not express TRPV1. are also unresponsive to changes in luminal pressure.. 5). 1997. 2004. Myers et al. 1996. These “cough receptors” are myelinated and do not synthesize and express neuropeptides under normal conditions (Riccio et al.. unlike RARs (Ho et al..

Microinjecting the antagonists 0.J. This is prevented by the sodium pump inhibitor ouabain. or bronchospasm (evoked by methacholine. histamine. The description of myelinated. 2003 for further details).8–1 mm rostral. The identification of the cough receptors as a distinct subtype of airway afferent nerve in guinea pigs may help clarify several contradictions between experimental evidence relating to cough and published interpretations of these results in which nomenclature was limited to the rigidly defined classes RARs. 6. 1999). Ouabain inhibits coughing evoked electrically. The existence of the cough receptors does. Terminals are labeled intravitally in the guinea pig trachea and bronchi using the styryl dye FM2-10 and visualized using epifluorescence microscopy in wholemounts pinned flat with mucosal side up. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 Fig. substance P or LTD4. It seems important to define the CNS terminations of the cough receptors to better differentiate these afferents from other airway afferent nerve subtypes. route of projection to the rostral trachea (recurrent laryngeal nerve). and recent immunohistochemical analyses indicate that the cough receptors may express the Na+ –K+ –2Cl− cotransporter NKCC1 (Mazzone. Coughing evoked by electrically stimulating the tracheal mucosa or by mechanically probing the laryngeal mucosa is also abolished in these animals. The stereotypical orientation of the cough receptors in the airway wall. however. Our previous dual tracing studies identified locations in nucleus tractus solitarius (nTS) near obex as termination sites of airway afferents arising from the nodose (and jugular) ganglia (Mazzone and Canning. Fuller. 7). Intravital labeling of the peripheral terminals of cough receptors innervating the mainstem bronchial mucosa of guinea pigs. vibrations or movement . 2006). lateral or caudal to this location is without effect on cough. These terminals form a complex arbor. distinct from that of tracheal and bronchial C-fibers. More recent (unpublished) studies have identified a discrete location in nTS. Canning et al. and unique morphology. The cough receptor terminals are brilliantly and selectively labeled by FM2-10. Immunohistochemical analyses in fixed tissues reveals that the cough receptors uniquely express an isozyme of the sodium pump containing the 3 subunit. 0–1 mm lateral from midline and 0–1 mm rostral to obex (defined here as the caudal most aspect of area postrema) that may contain the CNS terminations of the tracheal and laryngeal cough receptors (Fig. mechanically or chemically (acid) from the tracheal mucosa (see Mazzone and Canning. Employing tracing methods along with microinjection and cough studies in anesthetized guinea pigs.. FM2-10 labeling is prevented at 4 ◦ C. pulmonary embolism. Microinjecting a combination of NMDA and non-NMDA glutamate receptor antagonists into this location abolishes citric acid evoked coughing in anesthetized guinea pigs while having little if any effect on basal respiratory rate. As in a spider web. call for a refinement of network models of the cough reflex that has not incorporated the existence of such an afferent nerve subtype. 2002). in press). this may now be possible. maximal inspiratory and expiratory efforts against a closed glottis. Cl− may play an important role in regulating cough receptor excitability (Canning et al. Their identity as the cough receptors was determined by their ganglionic origin (nodose ganglia). acid and mechanically sensitive afferents innervating the extrapulmonary airways and regulating cough makes it unnecessary to explain why well-defined stimulants of intrapulmonary RARs (lung collapse. as rats do not possess a cough reflex. Identification of the peripheral terminals of the cough receptors may also shed light on the transduction of cough in health and in disease. Rewarming the tissue after thorough washing of excess dye results again in brilliant and selective labeling of the cough receptors. for example)) do not reliably evoke coughing. with their circumferential arbor and apparent adherence to the extracellular matrix of the basement membrane suggests that the ‘receptive field’ of these mechanoreceptors includes the nerve terminals but also the associated matrix. This cannot be carried out in rats. with preterminal and terminal branching and a circumferential alignment around the airways between the smooth muscle (faintly visible in panel C) and epithelium (which must be removed to visualize but not to stain the cough receptors). SARs and C-fibers.234 B.

Voltage sensitive dyes and perhaps even patch clamp analyses may also be used to characterize the excitability of the cough receptor terminals. activation of airway Cfibers evokes axon reflex-dependent peripheral release of the neuropeptides substance P. Axon reflexes in the airways and lungs induce bronchospasm. which is often associated with airways disease (Jeffery. That a single. leukocyte recruitment. 1990..5–1 mm lateral.. 1996.. so the results were pooled. Canning et al. 1993. Canning et al. 1995. n = 12) are depicted. loss of the epithelium may increase access to these terminals for tussigenic stimuli. Interactions between afferent nerves subtypes evoking cough 4. the antagonists reduced respiratory rate and reduced or abolished coughing from several of the injection sites studied. Joad et al. 2001). cumulative coughs evoked by citric acid in anesthetized guinea pigs after microinjecting vehicle (open squares.. vasodilatation. Immunohistochemical analyses (e. altered parasympathetic nerve activity and stimulation of endothelial and epithelial cells (Lundberg et al. Peripheral interactions In guinea pigs and rats. 1999. . n = 4). In the graph. Kuo et al. 1983.2 nmol each). Manzini. CNQX and AP-5 (0. 1997. The dissociation of the cough receptors from the smooth muscle and the dependence of the cough reflex on sustained (≥1 s) high frequency activation (upwards of 8 Hz) of multiple units may explain why the slowly developing and distant process of smooth muscle contraction and the resulting bronchospasm is a poor stimulus for cough (but may modify the cough reflex or cough sensitivity). rostral.02 nmol) had little or no effect on basal respiratory rate.8 mm lateral to obex (shown in figure inset and marked bilaterally with asterisks (scale bar: 1 mm)).5–1 mm rostral and 0.. 1997). 4. Cough was subsequently evoked by citric acid (0. NKA and CGRP (Barnes. In the diseased airways. This is now feasible when used in combination with intravital labeling techniques such as FM2-10. Piedimonte et al.1. or perhaps secondary to turbulent airflow may be a primary stimulus for cough. When microinjected in combination at higher concentrations (0.02 nmol CNQX and 0. Neuropeptide release in the airways or exogenously administered substance P activates RARs in rabbits and in guinea pigs (Bonham et al. while the airways obstruction and mechanical consequences of accumulated mucus and bronchospasm may initiate or increase the probability of coughing. At the concentrations administered.02 nmol AP-5 into various locations in nTS. 2001). may also alter cough receptor excitability.. Baluk et al. A discrete location in nTS plays an essential role in regulating coughing evoked by citric acid applied topically to the tracheal mucosa of guinea pigs. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 235 Fig. 2002). The neuropeptide-evoked RAR activation probably occurs secondary to actions on structural cells in the airway wall that in turn indi- within the matrix induced by impacting inhaled particulates or accumulated mucus. or microinjecting CNQX and AP-5 into adjacent locations in nTS (filled circles. Ionotropic glutamate receptors (both NMDA and non-NMDA subtypes) were blocked by simultaneously microinjecting (200 nL/injection) 0.001–2 M) applied topically to the tracheal mucosa in 100 L aliquots and at 1 min intervals.. Remodeling of the matrix. caudal and/or medial to the active site. mucus secretion. Vehicle (saline) was without effect regardless of the microinjection location. Matsumoto et al. Adjacent locations studied were 0. ion channel expression) focusing on these terminals within the airways may help identify novel therapeutic strategies for treating cough. resulting in a non-uniform deformation of the receptor may be the driving force behind mechanical sensitivity in these afferents. Cough was blocked most effectively at a site 0. edema. punctuate mechanical stimulation of the receptive field activates the cough receptors (10–20 Hz over a 1–4 s period) whereas changes in luminal pressure have no effect suggests that a directional and nonuniform force put upon the receptive field. Retrograde tracing confirmed that this nTS location is a site for tracheal afferent termination (not shown).J.B. punctate mechanical stimulation. 7. Ricciardolo et al. Bilateral injections were made into lateral nTS locations.. 1992.g. n = 17) or CNQX and AP-5 into the active site identified in the inset figure (open circle. The uniform physical forces associated with a luminal pressure change may be insufficient for cough receptor activation..

1985. 2001). 1996). 1982. 1997. 2001...c. 16 Hz. bronchospasm or the neutral endopeptidase inhibitor phosphoramidon (Kohrogi et al. 1998). 10 s train) from the extrathoracic tracheal mucosa of anesthetized guinea pigs were made with and without prior C-fiber stimulation. Pisarri et al.J. Canning et al. In all species. 1993. The results are shown as a percentage of the animals in each treatment group that coughed in response to 4 V stimulation of the trachea (see Mazzone et al. Attempts to evoke cough electrically (4 V. Bolser et al. Hansson et al. cigarette smoke. Endogenous neuropeptides and the axon reflex are unlikely to play a comparable role in the airways of species other than rats and guinea pigs (Barnes... 1993. C-fiber activation does not evoke cough in anesthetized guinea pigs but sensitizes the reflex to subsequent stimulation. 1984. 2001. as the sensitization was prevented by i. Ujiie et al. and was mimicked by microinjecting substance P into cnTS (Panel D). Ichinose et al. Sekizawa et al. The associated increases in RAR activity can be substantially reduced or abolished by pretreating animals with isoproterenol... (B) intrathoracic airway challenge with an aerosol of 1 mg/mL bradykinin (physically separated from the tracheal segment where cough was evoked). 2002. The sensitization of cough appears to be mediated by centrally released tachykinins.. 2002). 2001. Davis et al. 8.236 B. 2002). 1994. .. C-fiber activation evokes CNS-dependent reflexes (Russell and LaiFook. rectly activate RARs (Bergren.. or (C) microinjection of capsaicin (5 nmol) into the commissural subnucleus of nTS (cnTS). 1979. administration of either NK1 (CP99994. n = 5) receptor antagonists (panel A). Sant’Ambrogio and Widdicombe. 1997). Jia et al. Fuller et al. Roberts et al. 1987.. Coleridge and Coleridge.. Canning. thereby preventing the obstruction produced by these agents (Bergren. Lee and Pisarri. 1981. That inhaled anticholinergics have some antitussive properties in animals and in human subjects is consistent with this notion (Lowry et al. C-fibers were activated by (A) capsaicin applied topically to the tracheal mucosa.. Preventing the axon reflex with -agonist. Lamb and Sparrow. These end organ effects are mostly mediated by acetylcholine released from airway parasympathetic nerves and may activate RARs in the airway wall. however. 1989. Canning et al. inhaled neurokinin receptor antagonists or inhaled neutral endopeptidase (which enzymatically inactivates neurokinins and bradykinin) is effective at preventing cough evoked by capsaicin.. Yasumitsu et al. 1988.v. n = 5) or NK3 (SB223412.. 1 ms pulse duration. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 Fig. Microinjecting capsaicin laterally into the trigeminal nucleus was without effect on coughing. 1995. Canning. 2005 for more details). Capsaicinand bradykinin-induced stimulation of RARs in guinea pigs correlates with the increases in pulmonary insufflation pressure evoked by these agents.. 1995.

2004).B. since the central synergistic interactions are prevented entirely by neurokinin receptor antagonists administered intracerebroventricularly. As mentioned above. 2002). may promote coughing by facilitating synaptic transmission at relay neurons in the brainstem. there is good evidence that activation of extrapulmonary RARs and C-fibers can initiate cough upon activation. References Adams. Concluding remarks It is well established that vagal afferent nerves regulate coughing. It remains somewhat unclear. Ho et al. 2002). D. Irwin et al. Finally. Anatomical and functional studies have shown convergence of vagal afferents at sites of brainstem integration. Widdicombe. particularly in the nucleus of the solitary tract (nTS) (Kubin and Davies. Thus. Canning et al. L. Importantly. 2003). 1995.. / Respiratory Physiology & Neurobiology 152 (2006) 223–242 237 4. in the absence of C-fiber stimulation. Species differences may account for some of this uncertainty. Centrally administered neurokinin receptor antagonists prevent this sensitization of the cough reflex. E. It is imperative that future studies identify mechanisms of integration of afferent nerve input in the CNS. and the mechanisms by which afferent nerves are activated. Activation of C-fibers in the lung evokes profound increases in cholinergic tone in the airways by facilitating airway mechanoreceptor actions in the brainstem.B. Coleridge and Coleridge.. Jordan. 343–357. by administering substance P to the brainstem (Mazzone and Canning. J.. . In the absence of airway mechanoreceptor activity. Comparable interactions have been described in the airways of dogs (House et al. 2001. 2002). 70 (3). what relative role the identified afferent nerve subtypes play in mediating cough. The sensitizing effect of nociceptor stimulation can also be mimicked. Green. Regarding the specific subtypes of afferents identified in the airways and lungs and their role in cough. even during large lung inflations (Armstrong and Luck.. 2005) (Fig. Direct evidence for central interactions between airway C-fibers and RARs in the regulation of airway parasympathetic tone has been documented (Mazzone and Canning.A. C-fibers are ineffective at evoking reflex responses. Strong. Recent studies in guinea pigs and circumstantial evidence gathered from the existing literature suggest that a cough receptor quite distinct from either Cfibers or RARs may regulate coughing. in anesthetized guinea pigs. intrapulmonary RARs and C-fibers.2. 5. The central processing of C-fiber afferent nerve activity must therefore be integrated into a reflex pathway that is continually receiving input from airway mechanoreceptors. however. but the arguments against a role for C-fibers in cough become somewhat circular when it is also argued that putatively selective C-fiber stimulants evoke coughing in conscious animals only because they also activate RARs. a comparable interaction between cough receptors and C-fibers has been documented in studies of coughing (Mazzone et al. 1984.. the role of consciousness and perception in coughing. Central interactions Airway afferent nerve subtypes may interact to regulate airway defensive reflexes including cough.. Schertel. Mazzone and Canning. 8). Schneider. E. By contrast.J. Physiol. 1974. whereas C-fibers are typically quiescent. via central interactions with RARs or cough receptors. 1987. Our recent discoveries of the central terminations of the cough receptors will facilitate future studies of afferent nerve subtype interactions in the regulation of cough.R. Such studies may shed light on mechanisms by which extrapulmonary disorders such as rhinosinusitis and gastroespophageal reflux disease initiate chronic cough (Morice et al.. The facilitating effects of C-fibers on RAR reflex pathways in the brainstem appear to be mediated by tachykinins. C-fiber activation. all of these afferent nerve subtypes may interact to produce cough and heightened sensitivity to tussive stimuli. Resp. and SARs throughout the airways and lungs likely play an unessential role in cough in any species.. but it is also possible that multiple pathways for cough exist. Respiratory reflexes in the anesthetized miniature swine. lung mechanoreceptors are sporadically active throughout the respiratory cycle. 2001. The sensitivity of the presumably C-fiber-dependent cough reflex to anesthesia has led to some confusion and disagreement about the role of these afferents in regulating cough.F. C-fiber activation does not evoke cough but greatly sensitizes the cough reflex evoked by activating the cough receptors. 2006). 2004...

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