Role of the solitary tract nucleus in mediating nociceptive evoked cardiorespiratory responses

We compared the cardiorespiratory reflex responses evoked by noxious stimulation of the forelimb and cornea. Due to the depressant effects of anaesthesia on visceral reflexes we compared data from an unanaesthetised decerebrate rat model--the working heart-brainstem preparation (WHBP), with the anaesthetised rat. In both experimental models stimulation of the forelimb (mechanical pinch) evoked a tachycardia (WHBP: 19 +/- 2 bpm) and a decrease in respiratory cycle length (WHBP: from 4.1 +/- 0.2 to 2.3 +/- 0.1 s). The magnitude of response in anaesthetised animals depended on anaesthetic depth. Mechanical stimulation of the cornea evoked a bradycardia (-49.2 +/- 4.8 bpm) and an increase in respiratory cycle length from 4 +/- 0.36 to 5.88 +/- 0.2 s which was only present in the WHBP. In the WHBP activation of forelimb and corneal nociceptors both elicited significant pressor effects; in anaesthetised rats there were inconsistent changes in arterial pressure. To determine a role for the nucleus of the solitary tract (NTS) in mediating nociceptive evoked responses in the WHBP, synaptic transmission was blocked reversibly following bilateral microinjections of cobalt chloride. The heart rate responses evoked from either forelimb or corneal nociceptors were attenuated by approximately 50% (P < 0.05). A similar effect was observed using isoguvacine, a GABAA receptor agonist, to hyperpolarise NTS neurones. In conclusion, activation of forelimb and corneal nociceptors evoked contrasting patterns of cardiorespiratory response in the WHBP while in the anaesthetised rat the magnitude of the cardiorespiratory response to forelimb stimulation was quantitatively dependent on anaesthetic dose. In the WHBP, NTS neurones appear important for mediating the cardiac component of the reflex response following stimulation of nociceptive reflex pathways.     

Cardiorespiratory reflexes in a working heart-brainstem preparation of the house musk shrew, Suncus murinus

In this study, we adapted the working heart-brainstem preparation (WHBP) from rodents to the Insectivore, Suncus murinus. Suncus WHBPs had a baseline heart rate of 333 +/- 8 beats min(-1), a perfusion pressure of 69 +/- 2 mm Hg and a respiratory cycle length of 6.5 +/- 0.7 s. Administration of atropine produced an increase in heart rate of 26 +/- 9 beats min(-1) indicative of the presence of cardiac vagal tone. Activation of baroreceptors produced pressure-dependent reflex falls in heart rate and reduced respiratory cycle length. The baroreceptor reflex sensitivity in Suncus WHBP was a decrease in heart rate of 8.1 +/- 1.4 beats min(-1) mm Hg(-1). Activation of peripheral chemoreceptors with aortic injections of sodium cyanide (0.1-12.5 microg) produced a dose-dependent reflex fall in heart rate and reduced respiratory cycle length. The reflex falls in heart rate evoked by baroreceptor and peripheral chemoreceptor stimulation were both atropine-sensitive. We conclude that viable WHBP can be prepared from Suncus and that Suncus WHBP is a novel non-rodent model in which to study brainstem-mediated reflexes.     

Importance of neurokinin‐1 receptors in the nucleus tractus solitarii of mice for the integration of cardiac vagal inputs

Unmyelinated vagal afferents from the heart terminate within the nucleus tractus solitarii (NTS) located in the dorsomedial medulla. The neurotransmitter and postsynaptic receptors mediating information from cardiac vagal receptors to the NTS are unknown. This study determined the effects of neurokinin-1 (NK₁) receptor blockade on: (i) the reflex response evoked following aortic root injection of either veratridine (1–3 μg/kg) or bradykinin (80–300 ng/kg) to stimulate cardiac receptors in in vivo anaesthetized mice; and (ii) the evoked synaptic response of cardioreceptive NTS neurons following both intraleft-ventricular injection of veratridine or bradykinin, and electrical stimulation of the ipsilateral vagus nerve in an arterially perfused working heart-brainstem preparation of mouse. Administration of CP-99,994 (0.75–1.5 mg/kg i.v.), a specific NK₁ antagonist, attenuated significantly the evoked reflex bradycardia and depressor response following cardiac receptor (n = 6), but not pulmonary chemoreflex stimulation in vivo. From extracellular recordings of cardioreceptive NTS neurons, CP-99,994 reduced reversibly the total number of evoked spikes, peak firing frequency and response duration evoked by intraventricular injections of veratridine (n = 5) or bradykinin (n = 5). The number of evoked action potentials following electrical stimulation of the vagus nerve was also reduced. In five whole cell recordings of NTS neurons, both the evoked depolarization following cardiac receptor stimulation, and the peak amplitude and duration of vagus nerve-evoked EPSPs were reduced by CP-99 994; synaptic inputs from both peripheral chemoreceptors or pulmonary C-fibres were unaffected. These data support a selective involvement of NK₁ receptors in the transmission of cardiac vagal afferent inputs to NTS neurons integrating cardiorespiratory information.     

Neural organization of the viscero-cardiac reflexes

The reflex responses evoked in the postganglionic nerves to the heart were tested in chloralose-anaesthetized cats. Electrical stimulation of the A delta afferent fibres from the left inferior cardiac nerve evoked spinal and supraspinal reflex responses with the onset latencies of 36 ms and 77 ms respectively. The most effective stimulus was a train of 3-4 electrical pulses with the intratrain frequency of 200-300 Hz. Electrical stimulation of the high threshold afferent fibres (C-fibres) from the left inferior cardiac nerve evoked the reflex response with the onset latency of 200 ms. The C-reflex was present in intact animals and disappeared after spinalization. The most effective stimulus to evoke this reflex was a train of electrical pulses delivered at a frequency of 1-2 Hz with an intratrain frequency of 20-30 Hz. The most prominent property of the C-reflex was its marked increase after prolonged repeated electrical stimulation. We conclude that: (1) viscero-cardiac sympathetic reflexes may be organized at the spinal and supraspinal level; (2) viscero-cardiac sympathetic reflexes evoked by stimulation of the A delta and C afferent fibres from the left inferior cardiac nerve have different central organization.     

ANP potentiates nonarterial baroreflex bradycardia: evidence from sinoaortic denervation in rats

Previous findings indicate that atrial natriuretic peptide (ANP) enhances the reflex bradycardia arising from stimulation of cardiac mechanoreceptors and chemoreceptors, but not that from arterial baroreceptors. The present study tests this proposal by examining the effect of ANP on these reflexes in six chronically sinoaortic-denervated (SAD), and eight sham-operated (sham), conscious rats. Arterial baroreceptor-heart rate (HR) reflex function was examined by constructing full-range steady-state blood pressure (BP)-HR curves using alternating doses of pressor (methoxamine, 2-100 microg/kg) and depressor (nitroprusside, 1-50 microg/kg) agents. Nonarterial baroreceptor reflex function was assessed by the 'ramp' bradycardic response to the rapid BP rise after i.v. methoxamine (100 microg/kg bolus dose). The cardiopulmonary chemoreflex was evoked by i.v. injections of serotonin (1-20 microg/kg). These three tests were performed on each rat during infusions, in random order, of rat ANP (150 ng/kg/min i.v.) and saline vehicle. The ability of ANP to significantly enhance ramp reflex bradycardia was not diminished in SAD compared with sham rats (+54 +/- 12% vs. +42 +/- 15%, respectively). ANP also significantly enhanced cardiopulmonary chemoreflex bradycardia in both groups (+60 +/- 15% in SAD, +40 +/- 8% in sham). Neither the normal steady-state BP-HR response in sham rats nor the small residual response in SAD rats was enhanced by ANP (-1 +/- 7% in sham, -11 +/- 8% in SAD). We conclude that ANP enhances reflex bradycardias of nonarterial, probably cardiac mechanoreceptor, origin.     

Controlled bradycardia induced by nasal stimulation in the muskrat, Ondatra zibethicus

Respiration was disrupted and bradycardia induced in anesthetized muskrats by stimulating the nasal cavity with a stream of either water or various concentrations of ammonia vapors. When responses induced by either ammonia or water were compared, ammonia vapors were considered preferable because the responses could be maintained reliably through relatively rapid periods of stimulation, and the post-stimulus recovery of heart rate and respiration was more predictable. Moreover, the bradycardia induced in the first 5 s of stimulation by dilutions of ammonia vapors was graded. After injections of lidocaine were made into the nucleus tractus solitarius a profound bradycardia to ammonia stimulation persisted despite disruption of normal respiratory rhythms and an inhibition of the baroreceptor reflex induced by phenylephrine administration. These results show that ammonia vapors stimulating the nasal chambers effectively elicit cardio-respiratory adjustments in anesthetized muskrats and that the bradycardia may be controlled by varying the intensity of the peripheral stimulus. The trigeminal contribution for this is emphasized since the bradycardia persists after reversible blockade of the solitary complex. These data suggest that the trigeminal input to cardiac motorneurons is via relatively few synapses and is over circuits which run parallel to those modulating cardiac activity in response to chemoreceptors, baroreceptors and pulmonary afferent fibers.     

Attenuation of the carotid body chemoreflex during the stimulation of the posterior vermis in the anaesthetised rabbit

The purpose of this study was to investigate the effects of electrical and chemical stimulation of the sub-lobule IX-b of the cerebellar uvula on the cardiorespiratory responses evoked on stimulation of the carotid chemoreceptors in the anaesthetised and paralysed rabbit. Recordings of arterial blood pressure, ECG, heart rate and phrenic nerve activity were evaluated. Sub-lobule IX-b was activated electrically (1 ms, 20 microA, 100 Hz, 4-s train) and stimulated chemically with sodium glutamate microinjections (2 mM, pH=7.4 +/- 0.1) using bipolar concentric or multibarrelled microelectrodes. Carotid body chemoreflexes were evoked by the retrograde injection of sodium cyanide (0.1%, 0.1 ml) into the blood supply of the carotid body through a cannula inserted in the external carotid artery. The interaction of uvula stimulation and the carotid chemoreceptor reflex was studied by following the chemoreceptor stimulus with a stimulus to the uvula. Electrical and chemical stimulation of the uvula evoked a characteristic bradycardia and a depressor response together with a decrease in phrenic nerve activity. Carotid body stimulation evoked a decrease in heart rate accompanied by an increase of both arterial blood pressure and phrenic nerve activity. However, simultaneous delivery of the two stimuli resulted in a depressor response together with a decrease of phrenic nerve activity and an increased bradycardia. These data suggest that stimulation of the cerebellar uvula exerts an inhibitory control of the chemoreflex efficacy as far as respiratory activity and blood pressure is concerned.     

A functional study of uncrossed and crossed pulmonary afferent fibres in the cervical vagus nerves of the cat

The functional distribution of uncrossed and crossed pulmonary afferent fibres in the cervical vagus nerves has been studied in the anaesthetized cat using acute and chronic unilateral pneumonectomized preparations. The heart and lungs were sympathectomized routinely. The vagal afferent pathways of three pulmonary reflexes were investigated: the Hering-Breuer respiratory reflex, the lung inflation cardio-accelerator reflex, and the pulmonary chemoreflex. Inflation of the remaining lung caused temporary inhibition of inspiration. It also resulted in acceleration of the heart, but only when the background cardiac vagal tone was augmented. These respiratory and cardiac responses were abolished in most animals by ipsilateral cervical vagotomy; however, in some, a small response persisted and this was abolished by contralateral vagotomy. Stimulation of pulmonary C-fibre endings with right atrial injections of phenylbiguanide caused a reduction in respiration, bradycardia and systemic hypotension, responses which occurred with a latency of 2.9 +/- 0.15 s. They were mostly abolished by ipsilateral cervical vagotomy, but reduced responses persisted in a few animals. The residual responses were abolished by contralateral cervical vagotomy and by selective denervation of the lung. These results indicate that most afferent fibres subserving the three pulmonary reflexes studied run in the ipsilateral cervical vagus, representing the uncrossed pathway. Some afferent fibres, however, cross to the contralateral cervical vagus. Degenerative changes in cells of the contralateral nodose ganglion in chronic unilateral pneumonectomized animals support these findings.     

Stimulation of 5-HT2 receptors in the nucleus tractus solitarius enhances NMDA receptor-mediated reflex-evoked bradycardiac responses in the rat

The modulation by 5-HT2 receptors in the nucleus tractus solitarius of the reflex bradycardia evoked by stimulation of peripheral baroreceptors and cardiopulmonary chemoreceptors, and their possible functional interactions with local NMDA receptors, were investigated in pentobarbital- and urethane-anaesthetized rats, respectively. Microinjection of the 5-HT2 receptor agonist, 2,5-dimethoxy-4-iodoamphetamine (0.1-0.5 pmol), into the nucleus tractus solitarius elicited a dose-dependent hypotension and bradycardia. Bilateral microinjections at the same site of a subthreshold dose of 2,5-dimethoxy-4-iodoamphetamine (0.05 pmol) significantly enhanced the aforementioned reflex-evoked bradycardiac responses. In contrast, local bilateral microinjections of the NMDA receptor antagonist, 2-amino-5-phosphonopentanoic acid (500 and 1000 pmol), reduced, in a dose-dependent manner, both reflex-evoked responses. The facilitatory effect of 2,5-dimethoxy-4-iodoamphetamine upon these reflex-evoked bradycardiac responses was prevented by prior local microinjection of low doses of either the selective 5-HT2 receptor antagonist, ketanserin (10 pmol), or 2-amino-5-phosphonopentanoic acid (100 pmol), which, on their own, did not affect the reflex-associated bradycardia. These data suggest that 5-HT2 receptors within the nucleus tractus solitarius participate in a facilitatory modulation of the reflex control of heart rate, probably through functional interactions with local NMDA receptors.     

Reflex responses of sympathetic preganglionic neurones initiated by different cardiovascular receptors in spinal animals

We recorded the discharge of 66 single sympathetic preganglionic fibres isolated from the third and fourth left thoracic sympathetic rami communicantes of spinal vagotomized cats. Various mechanical and chemical stimuli were applied to different cardiovascular receptors in order to initiate reflex responses. Stimuli affecting mainly cardiac receptors always increased the discharge of the responsive fibres.Conversely, stimuli affecting simultaneously cardiac and vascular receptors or restricted to aortic receptors, either increased or decreased the discharge of the same fibres. The responses, consistent for each individual fibre, were reflex in nature. The data demonstrate that both specificity and convergence regulate cardiovascular sympathetic spinal reflexes.     

Reflex patterns in postganglionic vasoconstrictor neurons following chronic nerve lesions

Lesions of limb nerves in man may be associated with a variety of painful disorders with trophic changes described by the generic term 'reflex sympathetic dystrophy'. Our hypothesis is that pain and trophic changes are produced by an abnormal discharge pattern in postganglionic neurons supplying the limb (see refs. 3,24). In relation to this hypothesis, reflex patterns in postganglionic vasoconstrictor neurons supplying the skin (CVC) and the skeletal muscle (MVC) of the cat hindlimb were investigated at various times after a peripheral nerve lesion had been produced. These reflex patterns were compared with those in animals without nerve lesions (control preparations). The following lesions were made: cutting and ligating the superficial peroneal nerve (skin nerve) with subsequent neuroma formation, suturing the central stump of the superficial peroneal nerve to the peripheral stumps of muscle branches of the deep peroneal nerve, suturing the central stumps of muscle branches of the deep peroneal nerve to the peripheral stump of the superficial peroneal nerve, cutting and resuturing the superficial peroneal nerve, deafferentation of the whole hindlimb. The responses of vasoconstrictor neurons to stimulation of arterial chemoreceptors, arterial baroreceptors (cardiac rhythmicity of postganglionic activity) and cutaneous nociceptors were tested. In the animals with nerve lesions, the following groups of postganglionic vasoconstrictor neurons were analyzed: neurons projecting to the lesioned nerve, neurons projecting to hairy skin through an intact skin nerve (sural nerve) and neurons projecting to skeletal muscle through intact muscle nerves. In control preparations without nerve lesions, MVC neurons were excited by stimulation of arterial chemoreceptors and cutaneous nociceptors and inhibited by stimulation of arterial baroreceptors. Most CVC neurons were inhibited by stimulation of chemoreceptors and nociceptors and weakly inhibited by stimulation of baroreceptors. In animals with nerve lesions a and b, many CVC neurons in the lesioned nerves, as well as in the non-lesioned cutaneous nerve nearby, behaved in the same manner as MVC neurons. With respect to the control, this difference proved to be statistically significant. In preparations with lesions a, b and c, MVC neurons did not change their reflex patterns. After nerve lesions d and e, no major changes of reflex patterns were observed in CVC and MVC neurons. The inhibitory influence of arterial baroreceptors on CVC activity decreased in deafferented preparations (lesion e).(ABSTRACT TRUNCATED AT 400 WORDS)     

Differential modulation of withdrawal reflexes by a cannabinoid in the rabbit

Inhibition of spinal and trigeminal withdrawal reflexes by morphine and by the cannabinoid agonist HU 210 has been studied in anaesthetized and in decerebrated rabbits. In intact, pentobarbitone-anaesthetized animals, the jaw-depressor reflex (JDR) evoked by stimulation of the tongue, and the reflex elicited in the ankle flexor tibialis anterior (TA) by stimulation of the toes were inhibited to the same extent by morphine (1-30 mg kg(-1) i.v. cumulative). In spinalized, anaesthetized rabbits morphine depressed the JDR to the same level as in non-spinal preparations, but the effect of the opioid on the TA reflex was significantly reduced. All effects of morphine were reversed by naloxone (0.25 mg kg(-1), i.v.). In anaesthetised intact animals, HU 210 depressed the JDR at a dose of 100 nmol kg(-1) i.v. cumulative, reduced reflexes evoked in the knee flexor muscle semitendinosus (ST) by stimulation at the toes at a dose of 30 nmol kg(-1) i.v. cumulative, but had no consistent or significant effects on the TA reflex to toe stimulation. The same results were obtained in spinalized, anaesthetised animals. In decerebrated, spinalized rabbits with no anaesthesia, HU 210 (30 nmol kg(-1)) depressed both ST and TA reflexes evoked by toe stimulation. These data reveal that trigeminal and spinal withdrawal reflexes are equally sensitive to morphine provided the spinal cord is intact, suggesting that at least part of the action of systemic morphine is due to activation of descending inhibition. The present results also show for the first time that cannabinoid agonists can inhibit trigeminal withdrawal reflexes. HU 210 had differential effects on the three reflexes studied depending on the presence or absence of anaesthesia. This is the first occasion on which we have found pharmacological distinctions between withdrawal reflexes, and indicates that spinal sensorimotor processing is more heterogeneous than has been suspected previously.     

Inhibition of the bradycardic component of the von Bezold–Jarisch reflex and carotid chemoreceptor reflex by periaqueductal gray stimulation: involvement of medullary receptors

Stimulation of the dorsolateral periaqueductal gray matter (dlPAG) and the B3 cell group inhibits the cardiovagal component of the baroreflex in rats. Our aim was to determine whether the defence reaction induces similar modulatory effects on the cardiac response of the von Bezold–Jarisch reflex and the carotid chemoreceptor reflex. We examined the effects of dlPAG stimulation on the reflex bradycardia triggered by systemic administration of phenylbiguanide or potassium cyanide. Electrical and chemical stimulation of the dlPAG produced marked inhibition of the cardiovagal components of the von Bezold–Jarisch and the carotid chemoreceptor reflexes. In addition, as 5-HT₃, NK₁ and GABA_A receptor activation blocks cardiac reflex responses, we studied whether these receptors were involved in the dlPAG-induced inhibitory effects. We found that, after microinjection of granisetron (a 5-HT₃ receptor antagonist), bicuculline (a GABA_A receptor antagonist) and GR-205171 (an NK₁ receptor antagonist) into the nucleus of the solitary tract (NTS), reflex bradycardic responses were preserved during dlPAG stimulation. Finally, activation of the B3 region also inhibited both reflex bradycardic responses, and these effects were prevented by prior blockade of 5-HT₃ receptors in the NTS. The inhibitory effect of dlPAG stimulation on the cardiac reflex responses was prevented by inhibition of neurons in the medullary B3 region. In conclusion, 5-HT₃, GABA_A and NK₁ receptors in the NTS appear to be involved in the inhibition of the von Bezold–Jarisch reflex and the carotid chemoreceptor reflex bradycardia evoked by activation of neurons in the dlPAG and the raphé magnus.     

Reflex responses to baroreceptor, chemoreceptor and nociceptor inputs in single renal sympathetic neurones in the rabbit and the effects of anaesthesia on them

Reflex responses of renal postganglionic neurones to stimulation of arterial baroreceptors, arterial and central chemoreceptors and cutaneous nociceptors, and the rhythmicity of their resting activity were studied in paralyzed, artificially ventilated rabbits, anaesthetized with either alfathesin or chloralose-urethane. A 'vasoconstrictor' response pattern was seen in all units. Perivascular balloon-induced falls in blood pressure increased firing while pressure rises silenced 90% of units and reduced firing in the rest. Resting activity was linked to pressure changes within the cardiac cycle and to the artificial respiratory cycle. The largest excitation occurred during hypoxia and injections of CO2 saturated solutions into the carotid artery while hypercapnia and stimulation of cutaneous nociceptors only slightly increased firing. Parameters characterizing rhythmicities and reflex responses were unimodally distributed with no apparent subgrouping of units on quantitative grounds. Unit response patterns were similar to those recorded in the whole renal nerve. With one exception, no silent units were found which responded to the afferent inputs studied. Nor was there a small-spike fibre group which was excited by angiotensin. However, reflex responses were significantly influenced by the anaesthetic regime selected for use. Under alfathesin, baroreceptor and chemoreceptor reflexes were double those found with chloralose-urethane. Under chloralose-urethane, hypoxia increased both rhythmicities, while under alfathesin, cardiac rhythmicity was decreased and respiratory rhythmicity was variably affected. We concluded that renal sympathetic neurones are a functionally uniform population which behave like vasoconstrictors.     

Characteristics of sympathetic reflexes evoked by electrical stimulation of phrenic nerve afferents.

In chloralose-anaesthetized cats, sympathetic reflex responses were recorded in left cardiac and renal nerve during stimulation of afferent fibres in the ipsilateral phrenic nerve. In cardiac nerve, a late reflex potential with a mean onset latency of 75.6 +/- 13.8 ms was regularly recorded which, in 20% of the experiments, was preceded by an early, very small reflex component (latency between 35 and 52 ms). In contrast, in renal nerve only a single reflex component after a mean latency of 122.1 +/- 13.1 ms was observed. Bilateral microinjections of the GABA-agonist muscimol into the rostral ventrolateral medulla oblongata resulted in a nearly complete abolition of sympathetic background activity and in an 88% reduction of the late reflex amplitude with only small effects on the latency of the evoked potentials. Under this condition, an early reflex component was never observed to appear. After subsequent high cervical spinalization, the residual small potentials which persisted after bilateral muscimol injections were completely abolished and in cardiac nerve an early reflex potential with a mean latency of 45 +/- 10 ms was observed in all but one experiment. The early reflex was therefore referred to as a spinal reflex component which, however, is suppressed in most animals with an intact neuraxis. In the renal nerve a spinal response was only observed in one experiment after spinalization. The results suggest that sympathetic reflexes evoked by stimulation of phrenic nerve afferent fibres possess similar spinal and supraspinal pathways as previously described for somato-sympathetic and viscero-sympathetic reflexes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Somatosympathetic reflex in a working heart-brainstem preparation of the rat

The purpose of the present study was to examine the cardiorespiratory responses (CR) evoked by a somatosympathetic reflex (SSR) in the working heart-brainstem preparation (WHBP). Sprague-Dawley rats (75-100 g) were anesthetized with halothane, bisected sub-diaphramatically and decerebrated pre-collicularly (n = 15). The preparation was transferred to a recording chamber and perfused via the thoracic aorta with Ringer's solution containing an oncotic agent (Ficoll, 1.25%). SSR was activated by electrical stimulation (5 s) of the brachial nerve (0.5-40 Hz, 1-20 V, 0.1 ms) or the forelimb (0.5-40 Hz, 5-60 V, 2 ms). Stimulation at 40 Hz significantly increased heart rate (HR, 366 +/- 10 to 374 +/- 9 beats/min), systemic perfusion pressure (PP, 83 +/- 5 to 89 +/- 6 mmHg) and phrenic nerve discharge (PND, 0.4 +/- 0.1 to 1.4 +/- 0.3 Hz). Ganglionic blockade with hexamethonium (300 microM) eliminated the tachycardia and pressor response but did not alter the tachypnea to forelimb stimulation (n = 3). Transection of the brachial nerve plexus abolished the increase in PP and PND (n = 4). This indicates that a neural reflex mediated these responses. Spinal transection (C1-C2) completely abolished all responses indicating that they were mediated via a supraspinal pathway (n = 2). Based upon these findings, we conclude that activation of somatosensory afferent fibers in the WHBP evokes a programmed pattern of autonomic responses altering the activity-state of both the cardiovascular and respiratory systems. The WHBP provides a unique opportunity to investigate the medullary circuits and neuronal mechanisms that may be involved in coupling cardiorespiratory and somatomotor activity during locomotion/exercise.     

Characterisation of c-Fos expression in the central nervous system of mice following right atrial injections of the 5-HT3 receptor agonist phenylbiguanide

Cardiopulmonary receptors relay signals to the central nervous system via vagal and spinal visceral afferents. To date there are no detailed topographical studies in mice indicating the distribution of central neurones activated following stimulation of cardiopulmonary afferents. In anaesthetised mice, we injected the 5-HT(3) receptor agonist phenylbiguanide (PBG), a drug that is known to stimulate cardiopulmonary afferent C-fibres, into the right atrium of the heart and mapped c-Fos expression within specific regions of the central nervous system. Intra-atrial injection of PBG produced a reflex cardiorespiratory response including a pronounced bradycardia and a respiratory depression. Using immunohistochemical detection of the protein product of the immediate-early gene c-fos, we mapped the brain regions affected by cardiopulmonary 5-HT(3) receptor stimulation. Within the nucleus of the solitary tract (nTS) of PBG-injected mice, we detected an increased number of c-Fos-positive nuclei in the dorsolateral and gelatinous parts at the level of the area postrema (-7.48 mm bregma) but not at more rostral or caudal levels (-7.76, -7.20, -6.84 and -6.36 mm bregma) relative to vehicle-injected control mice. In addition, c-Fos expression in the crescent part of the lateral parabrachial nucleus was decreased in PBG-injected mice whereas no significant differences were detected between PBG-injected and control mice in the number of c-Fos-positive nuclei in the dorsal part of the lateral parabrachial nucleus. PBG injections had no significant effects on the number of c-Fos-positive catecholaminergic neurones within the C1/A1, C2/A2, A5, A6 and A7 cell groups. Likewise, PBG injections had no significant effects on c-Fos expression in other central regions involved in cardiorespiratory control or cardiorespiratory reflexes (selected non-catecholaminergic nuclei in the medulla and midbrain periaqueductal gray, the paraventricular nucleus of the hypothalamus and the central nucleus of the amygdala). Identification of specific regions of the nTS complex involved in relaying signals from afferent cardiopulmonary C-fibres to the central nervous system will be useful for future studies aimed at understanding neural mechanisms underlying cardiopulmonary reflexes and physiological responses to cardiopulmonary disease.     

Effects of a distant noxious stimulation on A and C fibre-evoked flexion reflexes and neuronal activity in the dorsal horn of the rat

In the halothane-anaesthetized rat, the responses of 49 neurons in the lumbo-sacral cord and the reflex discharge in the common peroneal nerve following electrical stimulation of the sural nerve were recorded in order to study possible relations between neuronal events and reflex nerve discharges. A distant noxious stimulus (to activate Diffuse Noxious Inhibitory Controls (DNIC) of Le Bars et al.) was used as a conditioning stimulus. Only the responses of neurons receiving an input from both A and C fibres were studied. The neurons were classified as class 1 (low threshold mechanoreceptive input only, n = 2), class 2 (nonnoxious and noxious inputs, n = 34) or class 3 (responding to noxious stimuli only, n = 13). During conditioning stimulation the C fibre evoked discharge was inhibited in 32 out of 34 class 2 neurons. The A fibre-evoked discharge was simultaneously inhibited in 29 of these neurons. The main effect of the distant noxious stimulation on the C fibre evoked neuronal discharge was to decrease the discharge by a constant number of spikes, independent of the level of evoked activity. Only one class 3 neuron was inhibited during conditioning stimulation and none of the class 1 cells were influenced by DNIC. During conditioning stimulation the late and prolonged C fibre evoked reflex nerve discharge (latency 160-200 ms, duration up to several hundred ms) was strongly depressed. Concomitantly, a short-lasting reflex nerve discharge appeared over the interval 115-160 ms. This released reflex nerve discharge (RR) had a constant latency. There was no simultaneous change of the A beta evoked reflex nerve discharge. After the end of the distant noxious stimulation the late C fibre evoked reflex nerve discharge (latency 160-200 ms) recovered. Concomitantly, the RR disappeared. The possibility that the class 2 neurons and the class 3 neurons are intercalated in different reflex pathways is discussed.     

Responses of vagal efferent fibres to stimulation of gastric mechano- and chemoreceptors in the anaesthetized ferret

The involvement of gastric tension receptors and mucosal chemoreceptors in vagal reflexes was assessed directly by recording single efferent fibres from the cervical vagus nerve in the urethane anaesthetised ferret. 32/39 fibres responded to distension of the gastric corpus, mainly with excitation of firing, 24/39 fibres responded to antral distension, with a higher proportion showing a reduction in firing. Fifty-seven percent of fibres showed a convergent input from corpus and antrum. The magnitude of these responses was large (usually greater than 50%) and of short latency (less than 1 s). Responses to chemical perfusions in the antrum were seen in 14/31 efferent fibres, most of which showed excitation. Latency of response was variable (5-120 s) and may have been influenced by diffusion through the antral mucous barrier. These data indicate a complex arrangement of vagal reflexes involved in pre-pyloric regulation of gastric emptying by mechanical and chemical stimuli in the lumen.     

Participation of different groups of afferent fibers of mesenteric nerves in vasomotor reflexes]

The dependence of the magnitude and character of vasomotor reflexes on the amplitude of mesenteric nerve tetanic stimulation was studied in experiments with cats anesthetized with urethane and chloralose. A comparison of the stimulus amplitude reflex magnitude curves with the previously obtained data on electrical excitability of various groups of mesenteric nerves afferent fibres revealed that there are 3 groups of "vasomotor" afferents in mesenteric nerves: "fast" Adelta-fibres (conduction velocity above 8 m/s) the impulses of which evoke depressor or small pressor reflexes; "slow" Adelta-fibres (conduction velocity below 8 m/s) the impulses of which evoke pressor reflexes or interacting with impulses of lower-threshold "fast" Adelta-fibres, either decrease depressor reflexes evoked by impulses of these fibres or increase pressor reflexes evoked by these fibres impulses; C-fibres the impulses of which increase pressor reflexes evoked by impulses of "slow" Adelta-fibres.