Right Atrial Injection Of Capsaicin Research Articles

Activation of bitter taste receptors in pulmonary nociceptors sensitizes TRPV1 channels through the PLC and PKC signaling pathway.

Bitter taste receptors (T2Rs), a G protein-coupled receptor family capable of detecting numerous bitter-tasting compounds, have recently been shown to be expressed and play diverse roles in many extraoral tissues. Here we report the functional expression of T2Rs in rat pulmonary sensory neurons. In anesthetized spontaneously breathing rats, intratracheal instillation of T2R agonist chloroquine (10 mM, 0.1 ml) significantly augmented chemoreflexes evoked by right-atrial injection of capsaicin, a specific activator for transient receptor potential vanilloid receptor 1 (TRPV1), whereas intravenous infusion of chloroquine failed to significantly affect capsaicin-evoked reflexes. In patch-clamp recordings with isolated rat vagal pulmonary sensory neurons, pretreatment with chloroquine (1-1,000 µM, 90 s) concentration dependently potentiated capsaicin-induced TRPV1-mediated inward currents. Preincubating with diphenitol and denatonium (1 mM, 90 s), two other T2R activators, also enhanced capsaicin currents in these neurons but to a lesser extent. The sensitizing effect of chloroquine was effectively prevented by the phospholipase C inhibitor U73122 (1 µM) or by the protein kinase C inhibitor chelerythrine (10 µM). In summary, our study showed that activation of T2Rs augments capsaicin-evoked TRPV1 responses in rat pulmonary nociceptors through the phospholipase C and protein kinase C signaling pathway.

Mu-opioid receptors in the caudomedial NTS are critical for respiratory responses to stimulation of bronchopulmonary C-fibers and carotid body in conscious rats

We tested the hypothesis that mu-opioid receptors (MORs) in the caudomedial nucleus tractus solitarius (cmNTS) are important for the ventilatory responses to stimulation of bronchopulmonary C-fibers (PCFs), the carotid body-mediated hypoxia, and hypercapnia independent of the carotid body. First, we used immunohistochemistry to map MORs distribution in the caudal medulla. Then we compared the effects of intra-cmNTS microinjection of DAMGO (a MOR agonist) with or without a combination of CTAP (a MOR antagonist) on the ventilatory responses to: 1) right atrial injection of capsaicin (to stimulation of PCFs) and 2) acute hypoxia (HVR, to stimulate the carotid body) in awake intact rats; and 3) hypercapnia (HCVR) in the carotid body ablated rats. The cmNTS presented the highest MORs in the caudal medulla. Microinjection of DAMGO blocked the PCF-mediated apnea, attenuated HVR (70%) and HCVR (21%), while microinjection of CTAP+DAMGO failed to affect these chemoreflexes. Our data demonstrate a critical role of activation of cmNTS MORs in regulating these chemoreflexes and imply a presence of MORs in the synapse of the 2nd-order neurons receiving inputs from PCFs and the carotid body, and NTS chemosensitive neurons.

Hypersensitivity of vagal pulmonary C-fibers induced by increasing airway temperature in ovalbumin-sensitized rats.

Our recent study has shown that hyperventilation of humidified warm air (HWA) triggered cough and reflex bronchoconstriction in patients with mild asthma. We suggested that a sensitizing effect on bronchopulmonary C-fibers by increasing airway temperature was involved, but direct evidence was lacking. This study was carried out to test the hypothesis that HWA enhances the pulmonary C-fiber sensitivity in Brown-Norway rats sensitized with ovalbumin (Ova). In anesthetized rats, isocapnic hyperventilation of HWA for 3 min rapidly elevated airway temperature to a steady state of 41.7°C. Immediately after the HWA challenge, the baseline fiber activity (FA) of pulmonary C-fibers was markedly elevated in sensitized rats, but not in control rats. Furthermore, the response of pulmonary C-fibers to right atrial injection of capsaicin in sensitized rats was significantly higher than control rats before the HWA challenge, and the response to capsaicin was further amplified after HWA in sensitized rats (ΔFA = 4.51 ± 1.02 imp/s before, and 9.26 ± 1.74 imp/s after the HWA challenge). A similar pattern of the HWA-induced potentiation of the FA response to phenylbiguanide, another chemical stimulant of C-fibers, was also found in sensitized rats. These results clearly demonstrated that increasing airway temperature significantly elevated both the baseline activity and responses to chemical stimuli of pulmonary C-fibers in Ova-sensitized rats. In conclusion, this study supports the hypothesis that the increased excitability of these afferents may have contributed to the cough and reflex bronchoconstriction evoked by hyperventilation of HWA in patients with asthma.

INHALATION OF HYDROGEN SULFIDE (H2S) AUGMENTS THE PULMONARY REFLEXES

We have previously reported that H2S exerted a sensitizing effect on pulmonary vagal C‐fiber afferents in anesthetized, artificially ventilated SD rats. However, whether the sensitizing effect of H2S in turn enhances pulmonary reflexes remains unknown. In anesthetized, spontaneously breathing rats, inhalation of aerosolized sodium hydrosulfide (NaHS, a donor of H2S) caused no significant changes in the baseline breathing pattern. However, the pulmonary chemoreflexes, characterized by apnea, bradycardia and hypotension, triggered by right atrial injection of capsaicin or adenosine, were potentiated after the NaHS inhalation. This potentiating effect was eliminated by pretreatment with perineural capsaicin (a selective blockade of the conduction of vagal C‐fibers) or HC‐030031 (an antagonist of TRPA1 receptors), but not affected by pretreatment with perineural sham surgery or HC‐030031 vehicle. Furthermore, in awake, restrained guinea pigs, the cough reflexes elicited by inhalation of aerosolized capsaicin were also potentiated after the NaHS inhalation. In conclusion, H2S enhanced pulmonary reflexes, and probably resulted from its action on pulmonary C‐fibers. (MOST 103‐2320‐B‐038‐014‐)

Hydrogen sulfide induces hypersensitivity of rat capsaicin-sensitive lung vagal neurons: role of TRPA1 receptors

The sensitization of capsaicin-sensitive lung vagal (CSLV) afferents by inflammatory mediators is important in the development of airway hypersensitivity. Hydrogen sulfide (H2S) is an endogenous mediator inducing hyperalgesia through transient receptor potential ankyrin 1 (TRPA1) receptors located on nociceptors. We conducted this study to determine whether H2S elevates the sensitivity of rat CSLV afferents. In anesthetized, artificially ventilated rats, the inhalation of aerosolized sodium hydrosulfide (NaHS, a H2S donor) caused no significant changes in the baseline activity of CSLV afferents. However, the afferent responses to right atrial injection of capsaicin or phenylbiguanide and to lung inflation were all markedly potentiated after NaHS inhalation. By contrast, the inhalation of its vehicle or NaOH (with a similar pH to NaHS) failed to enhance the afferent responses. Additionally, the potentiating effect on the afferent responses was found in rats inhaling L-cysteine (a substrate of H2S synthase) that slowly releases H2S. The potentiating effect of NaHS on the sensitivity of CSLV afferents was completely blocked by pretreatment of HC-030031 (a TRPA1 receptor antagonist) but was unaffected by its vehicle. In isolated rat CSLV neurons, the perfusion of NaHS alone did not influence the intracellular Ca(2+) concentration but markedly potentiated the Ca(2+) transients evoked by capsaicin. The NaHS-caused effect was totally abolished by HC-030031 pretreatment. These results suggest that H2S induces a nonspecific sensitizing effect on CSLV fibers to both chemical and mechanical stimulation in rat lungs, which appears mediated through an action on the TRPA1 receptors expressed on the nerve endings of CSLV afferents.

Hypersensitivity of bronchopulmonary C‐fibers induced by an increase in airway temperature in ovalbumin (Ova)‐sensitized Brown Norway rats.

Our recent study has reported that hyperventilation of humid warm air (WA) triggered cough and reflex bronchoconstriction in patients with mild asthma (AJRCCM 2012). We suggested that sensitization of the bronchopulmonary C‐fibers by increasing airway temperature is involved, but direct evidence is lacking. This study was carried out to determine whether hyperventilation of WA enhanced the pulmonary C‐fiber sensitivity in an animal model of asthma (Ova‐sensitized Brown‐Norway rats). Isocapnic hyperventilation of WA for 3 minutes rapidly raised airway temperature to a peak of ~42°C which significantly elevated the base‐line fiber activity (FA) of pulmonary C‐fibers in Ova rats immediately after the termination of WA challenge, but not in the control rats. Furthermore, the pulmonary C‐fiber response to right atrial injection of capsaicin was significantly higher in Ova rats than control rats before the WA challenge; this difference in the FA response to capsaicin was further amplified after WA in Ova rats (Δ=4.98±1.78 imp/s before, and 10.34±2.21 imp/s after WA; P<0.05, n=9). A similar pattern of the WA‐induced potentiation of the FA response to phenylbiguanide was also founded in Ova rats. These results suggest that Ova‐sensitization increases the excitability of pulmonary C‐fibers, and the hypersensitivity is further enhanced by an increase in the airway temperature. (Supported by NIH and DoD grants)

Pulmonary Chemoreflex Responses Are Potentiated by Chronic Treatment of Tumor Necrosis Factor Alpha (TNFα) in Mice

TNFα, a pro‐inflammatory cytokine, is involved in the pathogenesis of asthma and also known for its sensitizing effect on somatic nociceptive afferents. This study was carried out to test whether chronic treatment of TNFα enhances pulmonary chemoreflex responses. Mice were divided into two matching groups (n=9 each), TNFα (10.08 μg/kg; 1μg = ~105 units) and vehicle were administered by intratracheal (i.t.) instillation in treated and control group, respectively, during a brief anesthesia. 24 h later, the dose‐related responses of apnea to right atrial injection of capsaicin (a TRPV1 activator; 0.25–1.0 μg/kg), allyl isothiocyanate (a TRPA1 activator; 0.3–1.2 mg/kg) and phenylbiguanide (a 5‐HT3 activator; 30–120 μg/kg) were all markedly elevated in TNFα‐treated group as compared to that in the control group. Furthermore, the elevated sensitivity was absent when the same TNFα treatment was applied to p55/p75‐null mice, in which both types of TNFα receptors were mutated. These results suggest a positive interaction between TNFα and pulmonary C‐fiber receptors, which is responsible for elevating the pulmonary chemoreflex sensitivity. (Supported by NIH grant HL96914)

N-Arachidonyl dopamine sensitizes rat capsaicin-sensitive lung vagal afferents via activation of TRPV1 receptors

We investigated the effect of N-arachidonyl dopamine (NADA), an endogenous agonist of both transient receptor potential vanilloid 1 (TRPV1) and cannabinoid CB1 receptors, on the sensitivity of rat capsaicin-sensitive lung vagal afferent (CSLVA) fibers. In artificially ventilated rats, an intravenous infusion of NADA (400 microg/kg/ml, 0.5 ml/min for 2 min) mildly elevated the baseline CSLVA fiber activity, whereas it markedly potentiated CSLVA fiber responses to a right atrial injection of capsaicin or adenosine, and to lung inflation. The potentiating effect on CSLVA fiber sensitivity to an adenosine injection or lung inflation was blocked by capsazepine pretreatment (a TRPV1 receptor antagonist), but was unaffected by AM251 pretreatment (a CB1 receptor antagonist). In spontaneously breathing rats, a NADA infusion similarly potentiated the CSLVA fiber-mediated apneic response evoked by an adenosine injection, and this potentiating effect was also prevented by capsazepine pretreatment. We concluded that NADA at the dose tested non-specifically increases CSLVA fiber sensitivity to chemical and mechanical stimulation via activation of TRPV1 receptors.

Sensitization of capsaicin-sensitive lung vagal afferents by anandamide in rats: role of transient receptor potential vanilloid 1 receptors

Anandamide (AEA), an arachidonic acid derivative produced during inflammatory conditions, is an endogenous agonist of both transient receptor potential vanilloid 1 (TRPV1) receptors and cannabinoid CB1 receptors. Sensitization of capsaicin-sensitive lung vagal afferent (CSLVA) fibers by chemical mediators is important in the pathogenesis of hyperreactive airway diseases. We investigated the effect of the intravenous infusion of AEA (2 mg x kg(-1) x ml(-1), 0.5 ml/min for 2 min) on the sensitivity of CSLVA fibers to chemical and mechanical stimulation in anesthetized rats. In artificially ventilated rats, AEA infusion only mildly elevated the baseline activity of CSLVA fibers. However, CSLVA fiber responses to right atrial injection of capsaicin, AEA, or adenosine and to lung inflation (tracheal pressure = 30 cmH(2)O) were all markedly potentiated during AEA infusion, which reverted 20 min after termination of the infusion. The potentiating effect on the sensitivity of CSLVA fibers to adenosine injection or lung inflation was completely blocked by pretreatment with capsazepine (a TRPV1 receptor antagonist) but was unaffected by pretreatment with AM281 (a CB1 receptor antagonist). In spontaneously breathing rats, right atrial injection of adenosine evoked an apneic response that is presumably mediated through CSLVA fibers. Similarly, the adenosine-evoked apneic response was potentiated during AEA infusion, and this potentiating effect was also completely prevented by pretreatment with capsazepine. These results suggest that AEA infusion at the dose tested produces a mild activation of TRPV1 receptors and this nonspecifically increases CSLVA fiber sensitivity to chemical and mechanical stimulation.

Ovalbumin sensitizes vagal pulmonary C-fiber afferents in Brown Norway rats

Sensitization of vagal lung C fibers has been postulated to contribute to the development of asthma, but support for this notion is still lacking. We investigated the characteristics and function of pulmonary C fibers (PCFs) in ovalbumin (OVA)-sensitized Brown Norway rats, an established animal model of asthma. Rats were sensitized with intraperitoneal injection of OVA or were treated with saline (control). In study 1, with the use of open-chest and artificially ventilated rats, inhalation of 5% OVA aerosol evoked an augmented increase in total lung resistance in the OVA-sensitized rats, compared with the control rats. Bilateral vagotomy or subcutaneous pretreatment with a high-dose of capsaicin for blocking of C-fiber function equally attenuated this augmented total lung resistance response, suggesting the involvement of PCFs. In study 2, with the use of anesthetized, spontaneously breathing rats, right atrial injection of capsaicin (1 microg/kg; a PCF stimulant) evoked an augmented apneic response in the OVA-sensitized rats, compared with the control rats. In study 3, with the use of open-chest, paralyzed, and artificially ventilated rats, the afferent PCF responses to right atrial injection of capsaicin (0.5 and 1.0 microg/kg), phenylbiguanide (8 microg/kg; a PCF stimulant), or adenosine (0.2 mg/kg; a PCF stimulant) were enhanced in the OVA-sensitized rats, compared with the control rats. However, the baseline activities of PCFs and their afferent responses to mechanical stimulation by lung hyperinflation in the OVA-sensitized and control rats were comparable. Our results suggested that OVA-sensitized Brown Norway rats possess sensitized vagal PCFs, which may participate in the development of the airway hyperreactivity observed in these animals.

Sensitizing effects of chronic exposure and acute inhalation of ovalbumin aerosol on pulmonary C fibers in rats

The effect of ovalbumin (Ova) sensitization on pulmonary C-fiber sensitivity was investigated. Brown-Norway rats were sensitized by intraperitoneal injection of Ova followed by aerosolized Ova three times per week for 3 wk. Control rats received the vehicle. At the end of the third week, single-unit fiber activities (FA) of pulmonary C fibers were recorded in anesthetized, artificially ventilated rats. Our results showed the following: 1) Ova sensitization induced airway inflammation (infiltration of eosinophils and neutrophils) and airway hyperresponsiveness in rats; 2) baseline FA in sensitized rats was significantly higher than that in control ones; 3) similarly, the pulmonary C-fiber response to right atrial injection of capsaicin was markedly higher in sensitized rats, which were significantly amplified after the acute Ova inhalation challenge; and 4) similar patterns, but smaller magnitudes of the differences in C-fiber responses to adenosine and lung inflation, were also found between sensitized and control rats. In conclusion, Ova sensitization elevated the baseline FA and excitability of pulmonary C fibers, and the hypersensitivity was further potentiated after the acute Ova inhalation challenge in sensitized rats. Chronic allergic inflammatory reactions in the airway probably contributed to the sensitizing effect on these lung afferents.

Sensitivity to capsaicin is induced in pulmonary rapidly adapting receptors (RARs) by ovalbumin (Ova)‐sensitization in Brown‐Norway rats

The effect of airway sensitization to Ova on the response of vagal pulmonary rapidly adapting receptors (RARs) to capsaicin was investigated in Brown-Norway rats. Two groups of young rats (sensitized and control) were exposed to aerosolized Ova and vehicle, respectively, 3 days a week for 3 weeks. Differential cell count of the bronchoalveolar lavage indicated a distinct increase in the numbers of eosinophils and neutrophils in the Ova-sensitized rats. Single-unit fiber activities (FA) of RARs were recorded in anesthetized, artificially ventilated rats, and identified by their firing behaviors and adaptation index of >70%. Our results showed that right atrial injection of capsaicin (0.5 and 1.0 μg/kg) evoked either negligible or no response in RARs of control rats (ΔFA = 2.3 ± 0.8 imp/sec; n=5), which was consistent with that previously reported in the rat (Ho et al., Respir. Physiol. 127: 113–124, 2001). However, in striking contrast, the same dose of capsaicin evoked an immediate stimulatory effect on RARs of Ova-sensitized rats (ΔFA = 6.7 ± 4.2 imp/sec; n=6), with distinct and intense responses in two of these receptors. These pilot data seem to suggest that Ova-sensitization in the airways induced an expression of capsaicin sensitivity in RARs, probably resulting from the chronic airway sensitization and inflammation in sensitized animals. (Supported in part by NIH HL 67379)

Epinephrine enhances the sensitivity of rat vagal chemosensitive neurons: role of β3-adrenoceptor

This study was carried out to determine whether epinephrine alters the sensitivity of rat vagal sensory neurons. In anesthetized rats, inhalation of epinephrine aerosol (1 and 5 mg/ml, 3 min) induced an elevated baseline activity of pulmonary C fibers and enhanced their responses to lung inflation (20 cm H(2)O, 10 s) and right atrial injection of capsaicin (0.5 microg/kg). In isolated rat nodose and jugular ganglion neurons, perfusion of epinephrine (3 microM, 5 min) alone did not produce any detectable change of the intracellular Ca(2+) concentration. However, immediately after the pretreatment with epinephrine, the Ca(2+) transients evoked by chemical stimulants (capsaicin, KCl, and ATP) were markedly potentiated; for example, capsaicin (50 nM, 15 s)-evoked Ca(2+) transient was increased by 106% after epinephrine (P < 0.05; n = 11). The effect of epinephrine was mimicked by either BRL 37344 (5 microM, 5 min) or ICI 215,001 (5 microM, 5 min), two selective beta(3)-adrenoceptor agonists, and blocked by SR 59230A (5 microM, 10 min), a selective beta(3)-adrenoceptor antagonist, whereas pretreatment with phenylephrine (alpha(1)-adenoceptor agonist), guanabenz (alpha(2)-adrenoceptor agonist), dobutamine (beta(1)-adrenoceptor agonist), or salbutamol (beta(2)-adrenoceptor agonist) had no significant effect on capsaicin-evoked Ca(2+) transient. Furthermore, pretreatment with SQ 22536 (100-300 microM, 15 min), an adenylate cyclase inhibitor, and H89 (3 microM, 15 min), a PKA inhibitor, completely abolished the potentiating effect of epinephrine. Our results suggest that epinephrine enhances the excitability of rat vagal chemosensitive neurons. This sensitizing effect of epinephrine is likely mediated through the activation of beta(3)-adrenoceptor and intracellular cAMP-PKA signaling cascade.

Respiratory syncytial virus infection in anesthetized weanling rather than adult rats prolongs the apneic responses to right atrial injection of capsaicin

Apnea is a common complication in infants infected by respiratory syncytial virus (RSV). A recent study has shown that intranasal inoculation of RSV in conscious weanling rats strengthens the apneic responses to right atrial injection of capsaicin (CAP), leading to 66% mortality. The objectives of the present study were to determine 1) whether RSV infection changes baseline minute ventilation (Ve) and arterial blood gases in anesthetized rats; 2) what the effects of RSV infection are on the respiratory responses to CAP; and 3) whether the RSV-strengthened apneic responses are age dependent. Our experiments were conducted in anesthetized and spontaneously breathing rats divided into four groups of weanling and adult rats that received either intranasal inoculation of RSV or virus-free medium. Two days after RSV infection (0.7 ml/kg), animal blood gases, baseline Ve, and Ve responses to right atrial injection of three doses of CAP (4, 16, and 64 microg/kg) were measured and compared among the four groups. Our results showed that RSV infection increased respiratory frequency (approximately 25%, P<0.05) in weanling but not adult rats, with little effect on arterial blood gases. RSV infection amplified the apneic responses to CAP in weanling but not adult rats, characterized by increases in the initial (40%) and the longest apneic duration (650%), the number of apneic episodes (139%), and the total duration of apneas (60%). These amplifications led to 50% mortality (P<0.05). We conclude that RSV infection increases respiratory frequency and strengthens the apneic responses to CAP only in anesthetized weanling but not adult rats.

Prostaglandin E2 enhances the sensitizing effect of hyperthermia on pulmonary C-fibers in rats

This study was carried out to investigate whether the pulmonary C-fiber hypersensitivity induced by hyperthermia is altered by prostaglandin E(2) (PGE(2)). Single-unit afferent activities of pulmonary C-fibers were recorded in anesthetized, artificially ventilated rats when the intrathoracic temperature (T(it)) was maintained at normal (N; approximately 36 degrees C) and hyperthermia levels (H; approximately 41 degrees C) by perfusion of heated saline into the thoracic chamber for 3min. After approximately 20min of recovery, the fiber activities were recorded again during infusion of PGE(2) at both N and H levels of T(it). Our study showed: (1) The baseline fiber activity and responses to lung inflation, right-atrial injection of capsaicin and adenosine were all increased by increasing T(it) from N to H, and these hyperthermia-induced increases in sensitivities were also significantly augmented by PGE(2). (2) These enhanced sensitivities induced by PGE(2) were abolished by pretreatment with AH6809 and AH23848, selective antagonists of EP(2) and EP(4) prostanoid receptors, respectively. In conclusion, the hyperthermia-induced hypersensitivity of vagal pulmonary C-fibers is potentiated by PGE(2), and this effect is mediated through activation of EP(2) and EP(4) prostanoid receptors.

Effect of PGE 2 on pulmonary C‐fiber response to hyperthermia in rats

Body temperature frequently increases to ~41°C during strenuous exercise or severe fever. Local tissue hyperthermia is known to occur during inflammatory reaction, and an increase in airway temperature has been reported in asthmatic patients (Am J Respir Crit Care Med. 165:181, 2002). A recent study in our lab (J Physiol. 565:295, 2005) showed intrathoracic hyperthermia induces the hypersensitivity of pulmonary C-fibers. In this study, we investigated whether the response of pulmonary C-fibers to hyperthermia is altered by prostaglandin E2 (PGE2), one of the major autacoids released in the lung during airway inflammation. Single-unit afferent activities of pulmonary C-fibers were recorded in anesthetized, artificially ventilated rats when the intrathoracic temperature (Tit) was maintained at normal (~36 °C; N) and hyperthermia levels (~41 °C; H) by the perfusion of heated saline into the thoracic chamber for 3 min. After 20 min of recovery, the fiber activities were recorded again during the perfusion of PGE2 (3 μg/kg/min, 3 min) at these two Tit, N and H. Our studies show: 1) During PGE2 perfusion, baseline fiber activities were higher than the control at both N and H levels of Tit. Furthermore, the hyperthermia-induced increase in baseline activity was also significantly augmented by PGE2 (P < 0.01). 2) The responses of pulmonary C-fibers to both lung inflation (15 and 30 cmH2O) and right atrial injection of capsaicin (1 μg/kg) during the perfusion of PGE2 increased in a similar pattern as that of the baseline. In conclusion, PGE2 markedly potentiates the hyperthermia-induced hypersensitivity of vagal pulmonary C-fibers to lung inflation and capsaicin. (Supported by NIH HL 67379)

Carotid sinus nerve is involved in cardiorespiratory responses to intracarotid injection of capsaicin in the rat

The carotid sinus nerve (CSN), important in cardiorespiratory modulation, mainly contains C fibers (CSCFs). Previous studies have demonstrated that selective stimulation of bronchopulmonary C fibers (PCFs) via right atrial injection of capsaicin (Cap; approximately 0.25 microg) results in an apnea (approximately 3 s) associated with hypotension and bradycardia. The present study was undertaken to determine the effects of activating CSCFs on cardiorespiratory activities. Intracarotid injection of Cap was performed before and after bilateral transection of the CSN in anesthetized and spontaneously breathing rats. Our results showed that 1) low doses of Cap (up to 2 ng) produced an increase in minute ventilation by elevating both tidal volume and respiratory frequency with the threshold dosage at 1.0 ng (P < 0.05); 2) high doses (4-64 ng) generated an apnea (prolongation of expiratory duration by approximately 8-fold) and hypertension (P < 0.05); 3) bilateral transection of the CSN reduced excitatory and inhibitory respiratory responses by 30 and 81%, respectively, and increased the hypertension by 88% (P < 0.05); and 4) the same doses of Cap delivered into the right atrium to stimulate PCFs failed to evoke detectable cardiorespiratory responses. Our results suggest that compared with PCFs, CSCFs are more sensitive to Cap stimulation and that activation of these fibers significantly modulates cardiorespiratory activity in anesthetized rats.

Ovalbumin sensitization alters the ventilatory responses to chemical challenges in guinea pigs

Patients with chronic bronchial asthma show a depressed ventilatory response to hypoxia (DVH), but the underlying mechanism remains unclear. We tested whether DVH existed in ovalbumin (Ova)-treated guinea pigs, an established animal model of asthma. Twelve guinea pigs were exposed to Ova (1% in saline) or saline aerosol (control) for 5 min, 5 days/wk, for 2 wk. After completing aerosol exposure, the animals were anesthetized and exposed to systemic hypoxia. Ova treatment had no effects on animal body weight, baseline cardiorespiratory variables, or arterial blood O2 and CO2 tensions, but it attenuated the ventilatory response to hypoxia (10 breaths of pure N2) by 65% (P < 0.05). When the animals were subjected to intracarotid injections of sodium cyanide (20 microg) and doxapram (2 mg) to selectively stimulate carotid chemoreceptors, the ventilatory responses were reduced by 50% (P < 0.05) and 74% (P < 0.05), respectively. In contrast, Ova exposure failed to affect the ventilatory response to CO2 (7% CO2-21% O2-balance N2 for 5 min; P > 0.05). Furthermore, the apneic response evoked by stimulating bronchopulmonary C fibers (PCFs) with right atrial injection of capsaicin (5 microg) was markedly increased in the Ova-sensitized group (5.02 +/- 1.56 s), compared with the control group (1.82 +/- 0.45 s; P < 0.05). These results suggest that Ova sensitization induces a DVH in guinea pigs, which probably results from an attenuation of the carotid chemoreceptor-mediated ventilatory excitation and an enhancement of the PCF-mediated ventilatory inhibition.

Hypersensitivity of pulmonary C fibers induced by adenosine in anesthetized rats.

Compelling clinical evidence implicates the potential role of adenosine in development of airway hyperresponsiveness and suggests involvement of pulmonary sensory receptors. This study was carried out to determine the effect of a low dose of adenosine infusion on sensitivity of pulmonary C-fiber afferents in anesthetized open-chest rats. Infusion of adenosine (40 microg x kg-1x min-1 i.v. for 90 s) mildly elevated baseline activity of pulmonary C fibers. However, during adenosine infusion, pulmonary C-fiber responses to chemical stimulants and lung inflation (30 cmH2O tracheal pressure) were markedly potentiated; e.g., the response to right atrial injection of capsaicin (0.25 or 0.5 microg/kg) was increased by more than fivefold (change in fiber activity = 2.64 +/- 0.67 and 16.27 +/- 3.11 impulses/s at control and during adenosine infusion, n = 13, P < 0.05), and this enhanced response returned to control in approximately 10 min. The potentiating effect of adenosine infusion was completely blocked by pretreatment with 8-cyclopentyl-1,3-dipropylxanthine (100 microg/kg), a selective antagonist of the adenosine A1 receptor, but was not affected by 3,7-dimethyl-1-propargylxanthine (1 mg/kg), an A2-receptor antagonist, or 3-ethyl-5-benzyl-2-methyl-4-phenylethynyl-6-phenyl-1,4-(+/-)-dihydropyridine-3,5-dicarboxylate (2 mg/kg), an A3-receptor antagonist. This potentiating effect was also mimicked by N6-cyclopentyladenosine (0.25 microg x kg-1 x min-1 for 90 s), a selective agonist of the adenosine A1 receptor. In conclusion, our results showed that infusion of adenosine significantly elevated the sensitivity of pulmonary C-fiber afferents in rat lungs and that this potentiating effect is likely mediated through activation of the adenosine A1 receptor.

Acute hypoxia prolongs the apnea induced by right atrial injection of capsaicin.

Inspiratory central drive is augmented by acute hypoxia that leads to a hyperventilation, but it is inhibited by capsaicin (Cap)-induced stimulation of pulmonary C fibers (PCFs) that produces an expiratory apnea. We hypothesized that acute hypoxia should shorten or eliminate the Cap-induced apnea. The ventilatory responses to bolus injection of Cap (0.2-0.5 microg) into the right atrium before and during acute hypoxia (10% O(2) for approximately 1 min; Hypoxia+Cap) were compared in anesthetized and spontaneously breathing rats. We found that Cap injection during acute hypoxia produced an extremely long-lasting apnea (69.67 +/- 11.97 s) that was 16-fold longer than the apnea induced by Cap alone (expiratory duration = 4.37 +/- 0.53 s; P < 0.01). A similar prolonged apnea was also observed during hypoxia in anesthetized guinea pigs. Bilateral vagotomy abolished apneic responses to Cap both before and during hypoxia. Subsequent recording of single-fiber activity of PCFs (PCF(A)) showed that acute hypoxia did not significantly affect baseline PCF(A) but that it doubled PCF(A) responses to Cap via increasing both the firing rate (3.34 +/- 0.76 to 7.65 +/- 1.32 impulses/s; P < 0.05) and burst duration (1.12 +/- 0.18 to 2.32 +/- 0.31 s; P < 0.05). These results suggest that acute hypoxia augments PCF-mediated inspiratory inhibition and thereby leads to an extremely long-lasting apnea. This interaction is partially due to hypoxic sensitization of PCF response to Cap.