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The aim of the present work was to identify the role of ATP acting on specific P2X receptors in the central mechanisms of thermoregulation. Immunohistochemical studies demonstrated that brainstem structures involved in controlling body temperature contained large number of nerve cells bearing P2X ATP receptors. Experiments on conscious rats involving intracerebroventricular administration of an ATP analog and P2X antagonists showed that both activation and blockade of central P2X receptors produced marked changes in body temperature. Analysis of the effects of these substances provided grounds for suggesting that ATP acting on P2X receptors fulfils an important function in the mechanisms of transmitting afferent information from peripheral thermal receptors to thermoregulatory centers in the brainstem with responsibility for heat loss, while in pyrogen-induced fever ATP acting on these receptors may be involved in the activity of the endogenous antipyretic system.

Transcripts for P2X2and P2X6subunits are present in rat CNS and frequently colocalize in the same brainstem nuclei. When rat P2X2(rP2X2) and rat P2X6(rP2X6) receptors were expressed individually inXenopusoocytes and studied under voltage-clamp conditions, only homomeric rP2X2receptors were fully functional and gave rise to large inward currents (2–3 μA) to extracellular ATP. Coexpression of rP2X2and rP2X6subunits inXenopusoocytes resulted in a heteromeric rP2X2/6receptor, which showed a significantly different phenotype from the wild-type rP2X2receptor. Differences included reduction in agonist potencies and, in some cases (e.g., Ap4A), significant loss of agonist activity. ATP-evoked inward currents were biphasic at the heteromeric rP2X2/6receptor, particularly when Zn2+ions were present or extracellular pH was lowered. The pH range was narrower for H+enhancement of ATP responses at the heteromeric rP2X2/6receptor. Also, H+ions inhibited ATP responses at low pH levels (2+on ATP responses was unchanged. Thus, the rP2X2/6receptor is a functionally modified P2X2-like receptor with a distinct pattern of pH modulation of ATP activation and suramin blockade. Although homomeric P2X6receptors function poorly, the P2X6subunit can contribute to functional heteromeric P2X channels and may influence the phenotype of native P2X receptors in those cells in which it is expressed.

We have investigated the source of central adenosine important in modulating the cardiovascular response to hypothalamic defense area (HDA) stimulation in alpha-chloralose-anesthetized rats. Microinjections of an ecto-5'-nucleotidase inhibitor, alpha,beta-methylene ADP (alpha,beta-meADP), were made into caudal nucleus of the solitary tract (cNTS) and rostral ventrolateral medulla (RVLM), and its effects on HDA-evoked responses were observed. Stimulation of HDA evoked an increase in arterial pressure and a secondary rise in arterial pressure after stimulation ceased. There was also an increase in heart rate and hindlimb blood flow. alpha,beta-meADP had no effect on resting levels of arterial pressure, heart rate, and hindlimb blood flow when injected into the cNTS or RVLM. alpha,beta-meADP also had no effect on the HDA-evoked tachycardia and increase in muscle blood flow. However, alpha,beta-meADP reduced the primary increase in arterial pressure evoked by HDA stimulation when microinjected into the cNTS. In contrast, alpha,beta-meADP reduced the secondary increase in arterial pressure when microinjected into the RVLM. From these results, we suggest that at least part of the adenosine released centrally during HDA stimulation is derived extracellularly from ATP metabolism.

1. Whole-cell patch-clamp recordings were made from 142 visually identified rat dorsal vagal preganglionic neurones (DVMs). Applications of 5-hydroxytryptamine (5-HT, 20 microM, 2 min) elicited a slow depolarization (8.2 +/- 0.5 mV, n = 59) in 95% of the cells tested, accompanied by an increase in excitability. In (68%) of DVMs the depolarization was associated with an increase in apparent membrane resistance (Rmt 22.7 +/- 2.2%). These depolarizations and increases in Rm (14.3 +/- 2.6%, n = 8) were maintained in a medium which blocked synaptic transmission. 2. The response to 5-HT was associated with a reversal potential (Erev) of -91 +/- 1 mV at an extracellular K+ concentration (LK+]o) of 4.2 mM. This correlated well with the K+ equilibrium potential (Ek = -89 mV). 3. The depolarizing effect of 5-HT was attenuated by the 5-HT2A/2C receptor antagonists, ketanserin (1 microM), LY 53,857 (1 microM) and the 5-HT1A/2A receptor antagonist, spiperone (1 microM). The 5-HT1A receptor antagonist, pindobind 5-HT1A (5 microM), had no effect on the depolarizing response to 5-HT. 4. The effect of 5-HT was mimicked by the 5-HT2A/2C receptor agonist, alpha-methyl-5-HT (50 microM), the 5-HT1 receptor agonist, 5-carboxamidotryptamine (20 microM) and the putative 5-HT4 agonist, 5-methyoxytryptamine (5 microM). The selective 5-HT4 receptor antagonist, GR113808, had no effect on the depolarizing effect of 5-HT or 5-MEOT on DVMs. 5. The 5-HT3 antagonists, MDL 72222 (10 microM) and ICS-205-930 (1 and 10 microM), partially reduced the effect of 5-HT. The 5-HT3 receptor agonist, 2-methyl-5-HT (100-300 microM), excited a proportion of neurones tested (56%) by evoking a depolarizing and/or an increase in postsynaptic potentials (p.s.ps). 6. These results are consistent with direct, postsynaptic actions of 5-HT on DVMs via 5-HT2A receptors, being mediated, in part, by the reduction of K+ conductance.

The effects of ET-1 and ET-3 on ventilation and carotid chemosensory discharge have been studied in rats anaesthetised with pentotarbitone. Autoradiographic studies were also performed in vitro to investigate the binding of [125I]ET-1 to rat carotid body, nodose ganglion and brain stem. ET-1 caused a dose-related hyperventilation that was abolished by cutting both carotid sinus nerves. Recordings of chemosensory discharge from the carotid sinus nerve confirmed that ET-1 caused chemoexcitation. ET-3 had only slight effects. The hyperventilation evoked by ET-1 was antagonised by the ETA receptor antagonist FR139317, but responses to hypoxia (10% oxygen) and to cyanide were unaffected. [125I]ET-1 bound to the carotid body, the nodose ganglion and to the brain stem, particularly in the region of the nucleus tractus solitarii. ET-1 binding in the carotid body was displaceable by FR139317, which is consistent with the functional evidence for ETA receptors in the carotid body. The effects of ET-1 on ventilation, coupled with the presence of ET binding sites in areas involved in respiratory and cardiovascular regulation, is consistent with a physiological role for ET in the control of respiration, but our evidence suggests that ET is not crucial for chemotransduction in acute hypoxia.

1. The action of adenosine in the mediation of the cardiovascular changes associated with the defence reaction has been investigated in the rat using two A1 receptor antagonists. 2. Cumulative doses of 1,3 dipropyl-cyclopentylxanthine (DPCPX) (0.3-3 mg kg-1) and ethanol (0.03-0.25 ml) and bolus doses of DPCPX (3 mg kg-1) and 8-sulphophenyltheophylline (8-SPT) (20 mg kg-1) were given into alpha-chloralose, paralysed and artificially ventilated rats. Recordings were made of arterial blood pressure and heart rate. 3. Ethanol, the vehicle for DPCPX, failed to modify the magnitude of the defence response; however, cumulative doses of DPCPX produced a dose-dependent decrease in the HDA (hypothalamic defence area)-evoked increase in arterial blood pressure, accompanied by a similar fall in the magnitude of the evoked heart rate response. 4. The evoked rise in arterial blood pressure was reduced significantly by intravenous injection of DPCPX (3 mg kg-1) but not 8-SPT (20 mg kg-1), a purely peripherally acting adenosine antagonist. 5. These results suggest that adenosine acting at A1 receptors located in the central nervous system, is involved in the HDA-evoked pressor response. Whilst the site of action of the A1 receptors is not known, possible locations are discussed.

1. In order to assess the importance of the parabrachial nuclei in modulating cardiorespiratory activity, electric current or microinjections of glutamate were used to stimulate discrete regions of the parabrachial nuclei in anaesthetized rats. 2. Stimulation of cell bodies in the medial region of the parabrachial nuclei and in the Kölliker-Fuse nuclei, caused an expiratory facilitatory response. This consisted mainly of a decrease in respiratory rate as measured by observing phrenic nerve activity. 3. Stimulation of cell bodies in the lateral region of the parabrachial nuclei caused an inspiratory facilitatory response. This consisted mainly of an increase in respiratory rate. 4. At the majority of sites (16 out of 20) where changes in respiratory rate were elicited by glutamate injection or electrical stimulation an increase in blood pressure was observed. The coexistence of increases in blood pressure and heart rate indicates the presence of inhibition of the heart rate component of the baroreflex and/or an increase in cardiac sympathetic drive. 5. The expiratory facilitatory response was not evoked reflexly by the rise in blood pressure since it was still present after administration of guanethidine, which abolished the rise in blood pressure. 6. The interactions between the parabrachial nuclei and the medullary respiratory complex in eliciting these changes are discussed.

Activation of the cerebella posterior vermis (uvula) produces marked changes in cardiovascular performance, via pathways involving the parabrachial nucleus, nucleus of the solitary tract (NTS), and rostral ventrolateral medulla. Blockade of the baroreceptor reflex during uvula stimulation results from an inhibitory action on neurons within the NTS.

The effects of electrical stimulation of the posterior cerebellar vermis in anaesthetized, decerebrate and conscious animals are described, and include marked changes in blood pressure and heart rate and an inhibition of the baroreceptor reflex. These effects appear to be restricted to lobule IX, and can be duplicated by chemical stimulation, indicating that they are a genuine cerebellar phenomenon. The results of both neuroanatomical and neurophysiological experiments to investigate the pathways responsible for the effects are described, and these show there to be a direct projection of Purkinje cell axons to the parabrachial nucleus. Experiments designed to test a possible involvement of lobule IX in the alerting response have proved negative, and while lobule IX itself appears to have no role in conditioned cardiovascular responses, lesions of lobules VI and VII do result in a significant impairment of the acquisition of conditioned bradycardia in the rabbit.

1. In anaesthetized rats recordings were made from thirty-eight neurones in the rostral ventrolateral medulla (RVL) with spinal-projecting axons. Their responses to mechanical, thermal and/or electrical stimulation were examined as were the accompanying changes in arterial pressure. 2. Mechanical, thermal and electrical stimulation of either hindpaw at a strength that can be regarded as noxious produced a consistent rise in arterial pressure. RVL-spinal-projecting 'vasomotor' neurones were excited by the noxious mechanical and thermal (52 degrees C) stimulation at a latency that was shorter than that of the evoked pressor response. 3. Percutaneous electrical stimulation of either hindlimb extremity resulted in an early peak of excitation (fourteen out of fourteen), an early trough of inhibition (twelve out fourteen), and a later peak of excitation (two out of fourteen). This response pattern to stimulation of either limb was independent of which limb was activated, but contralateral hindpaw stimulation elicited excitation at a shorter latency. The differences in latency of responses to stimulating two locations along the tail suggested that the early excitation and inhibition of RVL-spinal 'vasomotor' neurones were evoked by activation of peripheral fibres with a mean conduction velocity in the A delta range. 4. Short-latency excitatory and inhibitory responses in RVL-spinal 'vasomotor' neurones were observed also when single-pulse stimuli were delivered within the lateral part of the spinal cord. 5. Ionophoretic application of bicuculline, a GABAA receptor antagonist, blocked the evoked inhibition of these neurones on electrical stimulation of the hindpaw without attenuating the excitatory input from the same stimulus. 6. These results indicate that RVL-spinal 'vasomotor' neurones receive an input from cutaneous nociceptive afferents. This suggests that these neurones mediate, at least partly, the cardiovascular responses related to nociceptor stimulation.

1. Electrical stimulation of lobule IXb of the posterior cerebellar vermis evoked an increase in heart rate and arterial pressure in the decerebrate rabbit. This pattern of response was followed by a rebound bradycardia which was maintained until arterial pressure had returned to control levels. Activation of the arterial baroreceptors during IXb stimulation failed to elicit reflex changes in heart rate and arterial blood pressure. 2. Extracellular recordings were made from eighty-one single neurones in the nucleus tractus solitarius (NTS), and adjacent structures, that received inputs from the carotid sinus nerve, aortic nerve and/or vagus. A proportion of these were characterized as baro- or chemosensitive by physiological stimuli. The nature of the input from lobule IXb onto these cells was studied subsequently. 3. In twenty-seven out of thirty-five barosensitive neurones, stimulation of lobule IXb either decreased or abolished on-going activity. The latency to onset of inhibition was 21 ms in fourteen cells. In a further twenty-one neurones the spikes evoked orthodromically by electrical stimulation of either the aortic or sinus nerves were either inhibited or severely reduced in number during short-train conditioning stimulation of lobule IXb. This effect was most potent when the cortical conditioning stimulus was timed to occur 25 ms before the aortic or carotid sinus nerves were stimulated. 4. It is suggested that stimulation of lobule IXb in the decerebrate rabbit expresses its influence on the cardiovascular system in part by an inhibitory action on neurones receiving baroreceptor inputs at the level of the NTS.

1. In this study we have examined the roles of the lateral parabrachial nucleus (lateral PBN) and nucleus tractus solitarius (NTS) as sites mediating the circulatory responses evoked from lobule IX b of the posterior cerebellar vermis in the decerebrate and anaesthetized decerebrate rabbit. Microinjection of either kainic acid or bicuculline into the lateral PBN and NTS was undertaken to assess the importance of neurones and GABAergic mechanisms, respectively, in expressing the cardiovascular responses evoked from lobule IX b. 2. In both the decerebrate and anaesthetized decerebrate rabbit activation of neurones within the lateral PBN with a microinjection of glutamate elicited tachycardia and a pressor response together with an increase in renal sympathetic nerve discharge and vasoconstriction in the hindlimb. A microinjection of GABA into the lateral PBN evoked an opposite pattern of response. Chemical lesioning of neurones within the rostral region of the lateral PBN or pharmacological blockade of GABAA receptor, abolished or significantly attenuated the cardiovascular response (bradycardia/depressor effect) evoked from lobule IX b in the anaesthetized decerebrate rabbit. 3. In the decerebrate animal the tachycardia/pressor response elicited from lobule IX b was suppressed when either kainic acid or bicuculline was microinjected into the caudal division of the lateral PBN. 4. A bradycardia, depressor response, an abolition of on-going renal nerve activity and vasodilatation in the femoral bed were evoked by activating neurones within a restricted region of the NTS with glutamate in the decerebrate and anaesthetized decerebrate rabbit. A microinjection of GABA into the NTS produced a qualitatively opposite pattern of response. The bradycardia/depressor response evoked from lobule IX b in the anaesthetized decerebrate rabbit was not altered significantly following a microinjection of either kainic acid or bicuculline into the ipsilateral NTS. However, in the decerebrate preparation following a microinjection of bicuculline into the ipsilateral NTS the tachycardia/pressor response evoked from IX b was either abolished or in many cases reversed to the pattern of response seen in the anaesthetized decerebrate rabbit. 5. The present experiments provide evidence for two functionally distinct pathways from the cardiovascular region of lobule IX b to the lateral PBN.(ABSTRACT TRUNCATED AT 400 WORDS)

The hypothalamus is a key area for the integration of the autonomic features of affective behavior. Hypothalamic defence area (HDA) stimulation evokes major cardiorespiratory changes as well as modifications of general autonomic activity both in the anesthetized and conscious animal. Micturition is due to an increase in pelvic parasympathetic activity and, in the cat, the anterior hypothalamus has been implicated in urinary bladder control with the demonstration of a dorsolateral vesicoconstrictor pathway and a ventromedial inhibitory pathway. In this study we have investigated the effect of electrical and chemical stimulation of the HDA on bladder pressure and contractions in rat. Female rats (n = 15) were anesthetized, paralyzed and ventilated artificially. Arterial blood pressure, heart rate, urinary bladder pressure and pelvic nerve activity were recorded. HDA was electrically (1 ms, 100 Hz, 5-10 s train at intensities up to 150 micro A) and chemically (sodium glutamate, 50 nl, 2mM) stimulated. For statistical analysis the t-test was used, data were expressed as mean +/- SEM. Values of t were taken as significant when p0.05.HDA stimulation at 100-150 micro A evoked changes of both mean blood pressure (mBP) and bladder pressure (BlP). However, stimulation at30 micro A allowed a distinction within HDA of two different regions, at the same antero-posterior and lateral level, but separated 100-150 micro m in depth, which evoked differential effects on blood pressure and urinary bladder pressure. Results show that low intensity stimulation at ventral sites evoked a significant increase of mBP (from 102 +/- 5.9 to 127 +/- 8.6 mmHg, n = 10, p0.0001) with little changes of BlP (from 12 +/- 2.2 to 16 +/- 2.9 cmH(2)O, n = 10, p0.0005), whilst at more dorsal sites significant increases of BlP were elicited (from 12 +/- 8.3 to 38 +/- 4.6 cmH(2)O,n = 10, p0.0001) with only a small rise of mBP (from 102 +/- 6.2 to 111 +/- 9.8 mmHg, n = 10, p0.005). Glutamate injections at dorsal sites evoked a rise of BlP (from 11 +/- 2.2 to 30 +/- 3.0 cmH(2)O (n = 5; p0.0001) with small changes in BP, whilst at ventral sites (n = 4) glutamate microinjections evoked changes in BP but not of BlP. In conclusion stimulation at different sites within HDA can elicit separate changes in BP and BlP.

The aim of the present work was to identify the role of ATP acting on specific P2X receptors in the central mechanisms of thermoregulation. Immunohistochemical studies demonstrated that brainstem structures involved in controlling body temperature contained large number of nerve cells bearing P2X ATP receptors. Experiments on conscious rats involving intracerebroventricular administration of an ATP analog and P2X antagonists showed that both activation and blockade of central P2X receptors produced marked changes in body temperature. Analysis of the effects of these substances provided grounds for suggesting that ATP acting on P2X receptors fulfils an important function in the mechanisms of transmitting afferent information from peripheral thermal receptors to thermoregulatory centers in the brainstem with responsibility for heat loss, while in pyrogen-induced fever ATP acting on these receptors may be involved in the activity of the endogenous antipyretic system.

We have examined the importance of the A5 region modulating cardiorespiratory responses evoked from the parabrachial complex (PB) in spontaneously breathing rats. Cardiorespiratory changes were analyzed in response to electrical stimulation and glutamate microinjections into the PB (10-20 nl, 1-2 nmol) before and after ipsilateral microinjection of muscimol (50 nl, 0.25 nmol) or lidocaine (50 nl, 0.5 nmol) within the A5 region. Stimulation of medial parabrachial and Kölliker-Fuse nuclei (mPB-KF) evoked a decrease in respiratory rate (P0.001) with a rise in blood pressure (P0.001) and heart rate (P0.05). After muscimol or lidocaine microinjections within the A5 region, the pressor and heart rate responses to mPB-KF stimulation were reduced (P0.05, both cases). Muscimol within the A5 region altered the respiratory response to glutamate stimulation of mPB-KF, evoking an increase in respiratory rate (P0.05). Lidocaine abolished the respiratory response to mPB-KF stimulation. Stimulation of the lateral parabrachial nuclei (lPB) caused an increase in respiratory rate (P0.001) with a rise in blood pressure (P0.001) and heart rate (P0.05). Muscimol or lidocaine microinjections within A5 region decreased heart rate (P0.05) and pressor responses (P0.05) evoked from lPB. The increase of respiratory rate persisted unchanged. To confirm functional interactions between A5 and PB, extracellular recordings of putative A5 neurones were obtained during PB stimulation. Eighty-three A5 cells were recorded, 35 were activated from the mPB-KF (42%). The results indicate that neurones of the A5 region participate in the cardiorespiratory response evoked from the different regions of the PB complex. The possible mechanisms involved in these interactions are discussed.

The aim of the present study was to elucidate the role of extracellular ATP acting on specific P2X receptors in the central mechanisms of thermoregulation. Using immunohistochemistry methods, it was found that the brainstem structures involved in the body temperature regulation contain a large number of nerve cells which possess P2X receptors for ATP. Experiments with intracerebroventricular application of a stable ATP analogue and P2X receptor antagonists to conscious rats showed that both activation and blockade of central P2X receptors resulted in marked changes in body temperature. Analysis of the effects of these agents suggests that ATP by acting on P2X receptors performs an important function in the mechanisms of transmission of afferent information coming from peripheral thermoreceptors to the brainstem thermoregulatory centres responsible for heat loss, and during pyrogen-induced fever can be involved in the action of the endogenous antipyretic system.

The Bezold-Jarisch reflex involves a marked increase in vagal (parasympathetic) efferent discharge to the heart. During micturition, there is a marked increase in parasympathetic activity to the bladder (pelvic nerve activity) and this coincides with periods of phasic bladder contractions. In recent studies, we have shown that regions of the CNS that modulate pelvic nerve activity also affect cardiovascular variables. Recent anatomical studies have shown neuronal connections between the areas of the CNS that modulate both reflexes. Accordingly, in the present study we investigate if during the Bezold-Jarisch reflex there are simultaneous changes in pelvic nerve activity.Experiments were performed in Wistar female rats, anaesthetised with alpha-chloradose i.p. (80 micrograms/Kg) supplemented i.v. as required. Arterial blood pressure, ECG heart rate, urinary bladder pressure and pelvic nerve activity were recorded.Stimulation of cardiac chemosensitive nerve endings with veratridine evoked a decrease in arterial blood pressure from 88.6 +/- 1.72 to 66.1 +/- 2.50 mmHg (n = 30; p0.0001) and of heart rate from 420.3 +/- 5.56 to 248.8 +/- 17.58 bpm (n = 13; p0.0001) There were simultaneous changes of pelvic nerve activity that preceded an increase in urinary bladder pressure from 10.2 +/- 0.28 to 26.3 +/- 1.52 cmH2O (n = 30; p0.0001) and bladder contractions. We believe that the reflex changes resulted from activation of cardiac chemosensory nerve endings, as equivalent injections of warm saline into the left ventricle produced no changes in either cardiovascular variables or pelvic nerve activity.These observations suggest a significant role for cardiac afferents in modulating bladder contractions. The integration and functional significance of these inputs remain to be elucidated. The clinical importance of this physiological result may contribute to a better understanding of the mechanism of some semiologic findings, notably the association of urgency to micturate with myocardial ischemia and myocardial infarction.

1. In in vivo experiments, DOI (a 5-HT(2) receptor agonist), MK-212 (a 5-HT(2C) receptor agonist), and BW-723C86 (a 5-HT(2B) receptor agonist) were applied by ionophoresis to neurones in the rat nucleus tractus solitarius (NTS) receiving vagal afferent input. 2. The majority of the putative 'monosynaptically' vagal activated cells were inhibited by both MK-212 (4/6) and DOI (2/4), but unaffected by BW-723C86 (12/14). In contrast, 'polysynaptically' activated NTS cells were excited by both BW-723C86 (13/19) and DOI (9/10). Inactive 'intermediate' cells were inhibited by BW-723C86 (9/12), MK-212 (5/6) and DOI (3/4), whilst active cells of this group were excited by BW-723C86 (7/13) and DOI (5/5). 3. The selective 5-HT(2B) receptor antagonist LY-202715 significantly reduced the excitatory actions of BW-723C86 on 'intermediate' and 'polysynaptic' cells (13/13), but not the inhibitory effects observed on inactive Group 2 cells (n=5) whereas the selective 5-HT(2C) receptor antagonist RS-102221 reversed the inhibitory effects of MK-212 and DOI on 'monosynaptic and 'intermediate' neurones. 4. Cardio-pulmonary afferent stimulation inhibited two of four putative 'monosynaptically' activated calls and all four inactive intermediate cells. These were also inhibited by DOI and MK-212. In contrast, cardio-pulmonary afferents excited all five active intermediate cells and all six putative 'polysynaptically' activated NTS cells, while all were also previously excited by BW-723C86 and/or DOI. 5. In conclusion, these data demonstrate that neurones in the NTS are affected differently by 5-HT(2) receptor ligands, in regard of their vagal postsynaptic location, the type of cardio-pulmonary afferent they receive and the different 5-HT(2) receptors activated.

We have investigated whether exogenously applied adenosine modulates neuronal activity in a region of the central nervous system crucial for cardiovascular regulation. Extracellular recordings were made from neurons in the rostral ventrolateral medulla of the anaesthetized rat. Ionophoretic application of adenosine altered ongoing activity in 91% of neurons, evoking either a long-lasting depression or a short-lasting increase in firing rate. Both responses were blocked by application of the broad spectrum adenosine receptor antagonist 8-sulphophenyltheophylline, indicating that the responses were mediated by specific cell surface receptors. The adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dimethylxanthine blocked the increase, and partially blocked the decrease in firing rate in response to adenosine. The GABA(A) receptor antagonist bicuculline also blocked the increase in firing rate in response to adenosine, suggesting that adenosine may inhibit release of GABA from axon terminals in this region. The adenosine A2a receptor agonist CGS 21680 produced a long-lasting depression of ongoing activity. These results suggest that A1 receptors mediate an increase in firing rate, whilst A1 and A2a receptors mediate decreases in firing rate in some rostral ventrolateral medulla neurons. Thus, adenosine has been shown to modulate the ongoing activity of neurons in the rostral ventrolateral medulla by acting at both A1 and A2a receptors. Accordingly, we suggest, and provide some evidence to support the idea, that adenosine acts as an important neuromodulator in this region of the central nervous system, possibly by modulating the presynaptic release of neurotransmitters such as GABA.

1. In current and voltage clamp, the effects of hypoxia were studied on resting and synaptic properties of hypoglossal motoneurones in barbiturate-anaesthetized adult cats. 2. Twenty-nine hypoglossal motoneurones with a mean membrane potential of -55 mV responded rapidly to acute hypoxia with a persistent membrane depolarization of about +17 mV. This depolarization correlated with the development of a persistent inward current of 0.3 nA at holding potentials close to resting membrane potential. 3. Superior laryngeal nerve (SLN) stimulation-evoked EPSPs were reduced in amplitude by, on average, 46% while IPSP amplitude was reduced by 31% SLN stimulation-evoked EPSCs were reduced by 50-70%. 4. Extracellular application of adenosine (10 mM) hyperpolarized hypoglossal motoneurones by, on average, 5.6 mV, from a control value of -62 mV. SLN stimulation-evoked EPSPs decreased by 18% and IPSPs decreased by 46% during adenosine application. 5. Extracellular application of the KATP channel blocker glibenclamide led to a blockade of a persistent outward current and a significant increase of SLN stimulation-evoked EPSCs. 6. We conclude that hypoglossal motoneurones have a very low tolerance to hypoxia. They appear to be under metabolic stress even in normoxia and their capacity to activate protective potassium currents is limited when compared with other brainstem neurones. This may help to explain the rapid disturbance of hypoglossal function during energy depletion.

1. Studies are described that indicate that hypothalamic stimulation, at sites eliciting the defence reaction, results in an increase in adenosine levels in both the nucleus tractus solitarius (NTS) and rostral ventrolateral medulla (RVLM) of the rat. 2. Adenosine receptor antagonists applied to these sites attenuate the pressor response elicited by hypothalamic stimulation. Adenosine appears to be produced extracellularly from ATP, which is released from axon terminals in both the NTS and RVLM. 3. The implications of these observations for the pharmacology of hypothalamic actions on reflex inputs that modify cardiorespiratory function are discussed.

Because the aortic nerve of the rat is believed to contain only barosensory fibers in a functionally significant number, stimulation of this nerve provides a convenient means to identify barosensitive neurons in the central nervous system. The aim of the present study was to describe the characteristics of responses to stimulation of the aortic nerve in the neurons of the ventrolateral medulla oblongata (VLM) from in vivo intracellular recordings. Data were obtained from 25 neurons that were either spontaneously active and exhibited pulse-synchronous discharge or were silent. Cells that showed a regular increase in their discharge rate during diastole responded with either an inhibitory postsynaptic potential (IPSP, group 1; n = 3) or a biphasic excitatory postsynaptic potential (EPSP)-IPSP (group 2; n = 10) to aortic nerve stimulation. Parameters of the inhibitory response were similar in both groups (27 +/- 2 and 31 +/- 2 ms for onset latency and 55 +/- 10 and 67 +/- 7 ms in duration, respectively). In group 2 cells, the inhibition was, however, preceded by a brief excitation (14 +/- 2 ms for onset latency and 13 +/- 1 ms in duration). Group 3 neurons, which showed a regular increase in their discharge rate during or before systole (n = 7), responded with an EPSP (20 +/- 2 ms for onset latency and 44 +/- 5 ms in average duration). Group 1 and 3 responses were also encountered in quiescent neurons (n = 5). Intracellular labeling revealed that neurons of all three types were localized within the rostral VLM. The functional significance of these findings is discussed.

1. The role of centrally located adenosine A1 receptors in the cardiovascular changes associated with the hypothalamic defence response has been investigated by in vitro autoradiography and the intraventricular application of an A1 receptor antagonist. 2. 8-Cyclopentyl-1,3-dipropylxanthine (DPCPX), a highly selective adenosine A1 antagonist and its vehicle, ethanol, were administered directly into the posterior portion of the fourth ventricle of alpha-chloralose anaesthetized, paralysed and artificially ventilated rats. 3. DPCPX (0.01 to 0.3 mg kg-1) caused a dose-dependent decrease in the magnitude of the evoked pressor response (from -13 to -23 mmHg) elicited on hypothalamic defence area stimulation at a dose 10 fold lower than that required to produce an equivalent effect following systemic administration whilst ethanol, the vehicle, had no effect. 4. In vitro autoradiography revealed a heterogeneous distribution of adenosine A1 binding sites in the lower brainstem of rats. Image analysis showed the ventrolateral medulla to have the highest density of A1 receptors. Intermediate levels of binding were seen in caudal regions of the nucleus tractus solitarii and the hypoglossal nucleus. 5. These data imply that a proportion of the cardiovascular response to hypothalamic defence area stimulation are produced by the activation of adenosine A1 receptors localized close to the surface of, or adjacent to, the fourth ventricle in the immediate vicinity of the injection site.

The primary role of the respiratory and cardiovascular systems in man (and mammals) is to ensure an adequate supply of oxygen to the tissues and removal of carbon dioxide from them. Respiration is regulated to maintain the arterial concentrations of oxygen, carbon dioxide and hydrogen ions within narrow physiological levels, whilst the performance of the cardiovascular system is modified continuously so that the pattern of peripheral blood flow is appropriate to ensure an adequate supply of oxygen to the tissues of the different organ systems [71]. The demands on the systems differ greatly in different behavioural circumstances (cf. Chap. 108). Obviously, respiratory status in terms of this primary objective depends on the integration of the control of the two systems so that the metabolic demands of the tissues can be served. In principle, the cardiovascular and respiratory systems could be controlled by independent mechanisms, providing that at some point they are integrated.

1. In anaesthetized, paralysed and artificially ventilated cats, recordings have been made in the nucleus tractus solitarii (NTS) to assess further the role of this nucleus in mediating the cardiorespiratory responses that are elicited on stimulation within the hypothalamic defence area (HDA). 2. The responses of NTS neurones to stimulation in the hypothalamus were assessed, as were their patterns of evoked response to electrical stimulation of the sinus (SN), aortic (AN), superior laryngeal (SLN) and vagus (VN) nerves. 3. Stimulation in the HDA affected the activity of 110 NTS neurones (85 studied in intracellular and 25 studied in extracellular recordings). The present study focused on those sixty-eight neurones that were excited by such stimulation (51 intracellular recordings and 17 extracellular recordings). 4. Of the sixty-eight neurones that were excited by HDA stimulation, seven revealed no changes in membrane potential or evoked discharge (2 neurones) but the stimulus facilitated the excitatory effects of stimulating either (or both) the SN and SLN. An additional group of neurones showed powerful excitatory responses to HDA stimulation (15 studied with extracellular and 35 with intracellular recording). Evoked EPSPs had onset latencies in the range of 1-30 ms. Of those thirty-five neurones displaying EPSPs, twenty-six were shown to receive convergent inputs on nerve stimulation. In nine neurones the early EPSP in response to HDA stimulation was followed by an IPSP. 5. In a further group of neurones HDA stimulation elicited a long-lasting IPSP, but this was not analysed further because its features have been described in detail in earlier studies from this laboratory. 6. The patterns of response of several neurones excited by stimulation in the HDA are consistent with them forming a group of NTS interneurones that mediate the hypothalamically evoked cardiovascular responses, including modulation of reflex function, which is a major feature of cardiorespiratory control. This possibility is discussed in the light of the present physiological observations and descriptions of an intrinsic NTS group of GABA-containing neurones that have been suggested to fulfil such a role.

1. This investigation was initiated because activation of laryngeal afferents, either by electrical stimulation of the superior laryngeal nerve (SLN) or by natural stimulation of receptors in the laryngeal mucosa, results in a cardiorespiratory response comprising bradycardia, hypotension and apnoea (phrenic nerve activity was suppressed). This pattern of response is qualitatively equivalent to the response that is evoked on activation of the arterial baroreceptors. 2. Preliminary studies indicated that the effects of activating the SLN were suppressed during stimulation in the hypothalamic defence area (HDA) at points that also blocked the effects of baroreceptor stimulation. 3. Recordings were taken from seventy-two neurones localized within the ipsilateral nucleus tractus solitarii (NTS) whose activity was modified by SLN stimulation. Sixty neurones responded with an EPSP on SLN stimulation; nine of these had an inspiratory firing pattern. Five neurones were seen to receive an IPSP on SLN stimulation. 4. Five respiratory SLN-activated neurones were unresponsive to stimulation of the other nerve inputs, whilst four received convergent EPSP inputs on sinus nerve (SN) stimulation. One cell of these four also received inputs from the aortic and the vagus nerves. Sixty-one non-respiratory SLN-activated neurones also received convergent inputs from the sinus nerve. Of these, fifty displayed an EPSP, four an IPSP and seven an EPSP-IPSP. Fifteen neurones also received inputs from the aortic nerve and seventeen from the vagus. 5. From the population of neurones affected by SLN stimulation, twenty-four of seventy were also influenced by HDA stimulation (3 were respiratory cells). Sixteen of these responses consisted of an EPSP (2 respiratory cells), five of an IPSP (1 respiratory cell) and three of an EPSP-IPSP. 6. In neurones receiving an IPSP on HDA stimulation, the SLN-evoked excitatory response was reduced throughout the period of HDA-evoked inhibition. These neurones were all shown to receive excitatory inputs from the arterial baroreceptors and laryngeal mechanoreceptors. 7. Additionally, in the thirty-seven neurones that were excited by SLN stimulation but received no direct synaptic input on HDA stimulation, a conditioning stimulus to the HDA evoked a block of SLN-evoked responses without an accompanying change in membrane potential. Several of these neurones were also affected by both baroreceptor and laryngeal mechanoreceptor stimulation. 8. These observations are discussed in the context of the role of the NTS in cardiorespiratory control. The potential importance of these interactions in respiratory distress are highlighted and the implications for the organization of central pathways for the control of autonomic and respiratory function are discussed.

1. There is evidence in the literature of a mutual facilitatory interaction between the arterial chemoreceptor reflex and the alerting stage of the defence reaction, particularly in relation to the patterning of cardiorespiratory activity. The present study has been designed to test the hypothesis that a portion of this interaction involves synaptic interactions within the nucleus tractus solitarii (NTS). 2. The study has involved an analysis of the effective interactions between the stimulation of the arterial chemoreceptors and the hypothalamic defence area (HDA) on the activity of NTS neurones recorded in anaesthetized, paralysed and artificially ventilated cats. 3. A group of eighteen NTS neurones was classified as chemosensitive, on the basis of displaying EPSPs on sinus nerve stimulation (SN) and their failure to show an excitatory response to baroreceptor stimulation. Thirteen of these neurones displayed pronounced excitatory responses to chemoreceptor stimulation. In sixteen of these neurones HDA stimulation elicited an EPSP; in four of these sixteen neurones this early EPSP was followed by an IPSP. In the remaining two (of 18) neurones HDA stimulation provoked no obvious synaptic response but facilitated the efficacy of both chemoreceptor inputs and SN stimulation. 4. Neurones shown to receive convergent inputs from the arterial chemoreceptors (and SN stimulation) and HDA, often displayed excitatory responses to stimulation of other peripheral inputs. Vagally evoked EPSPs were observed in nine neurones, SLN-evoked responses in seven neurones and aortic nerve-evoked EPSPs in three neurones. 5. The organization of these synaptic interactions is discussed and these data are used to explain the pattern of interaction between chemoreceptor, baroreceptor and HDA inputs within the NTS. Conclusions are drawn regarding the functional role of different classes of NTS neurone, based on the findings in this and the accompanying two papers.

There is a body of data that indicates that the generation of respiratory activity involves the interplay of a limited number of distinct classes of respiratory neurones in the ventrolateral medulla [1]. Similarly the cardiovascular system is regulated by the activity of premotor and preganglionic neurones localised in an equivalent region of the CNS [2]. Cardio-respiratory homeostasis depends on the integration of the activity of the two systems and it could be envisaged that this was achieved by the action of two independent control systems that are loosely dependent on one another. Studies over the last decade have drawn attention, however, to the fact that the two systems are tightly coupled being sensitive to the same reflex inputs and participating in the expression of behavioral activities in a totally co-ordinated manner [3]. This has led to a recognition that rather than operating through independent control systems the CNS acts to maintain cardio-respiratory homeostasis by means of a single group of‘ cardio-respiratory’ neurones located in the ventrolateral medulla which regulate the activity of the two systems. This is achieved by generating complementary patterns of discharge of bulbospinal neurones that control respiratory motoneuronal and sympathetic preganglionic neuronal firing and through brainstem interneurones influencing the activity of vagal motoneurones that supply the heart, lungs and airways (and subsidiary muscles of respiration). This notion is not novel being firmly based in the 19th century literature but the experimental justification is only now emerging [4]. This results from an extensive electrophysiological analysis of the properties of the various motoneurones and interneurones in these networks, the nature of the reflex inputs impinging on them and the action of regions of the forebrain in generating changes in their activity.

In pentobarbitone anaesthetized and paralysed cats the effects of the A1 adenosine antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) have been observed on the pressor response to stimulation of the hypothalamic defence area (HDA) and on the effects of baroreceptor and chemoreceptor reflex activation on arterial blood pressure. The administration of DPCPX decreased the magnitude of the HDA pressor response and the chemoreceptor induced rise in blood pressure. The fall in blood pressure induced by baroreceptor activation was enhanced to a small yet significant extent. No changes were observed in the tachypnoea evoked by hypothalamic defence area (HDA) stimulation. These results suggest a possible role for central adenosine A1 receptors in mediating the cardiovascular changes evoked during HDA stimulation.

The central distribution of vagal afferents in the medulla containing either substance P, calcitonin gene-related peptide or 5-hydroxytryptamine was examined using a double-labelling technique and laser scanning confocal microscopy. Areas of the nucleus tractus solitarii, dorsal motonucleus of the vagus nerve and area postrema were scanned for double-labelled axon profiles. Analysis of this material revealed that all three neurochemicals were contained within the central terminals of vagal nerve sensory neurons. However, the distribution of vagal nerve afferents containing each of these putative transmitters differed. Afferents containing 5-hydroxytryptamine were detected mainly in the areas postrema and the adjacent nucleus tractus solitarii, with a smaller number in the ventral subnuclei of the solitary tract. In contrast afferents containing calcitonin gene-related peptide were found primarily in the medial and commissural regions of the nucleus tractus solitarii. Afferents containing substance P-immunoreactivity were surprisingly few in number and did not appear to be associated with any particular region. These results establish the presence of 5-hydroxytryptamine, substance P and calcitonin gene-related peptide in the central axons of vagal sensory afferents. Furthermore, the differential distribution of afferents immunoreactive for these neurochemicals seen in this study, together with previous demonstrations of the viscerotopic organization of vagal sensory afferents suggests a possible “chemical coding” for individual end organs.

Investigations continue into the central neuronal circuitry involved in cardiovascular control, the activity of sympathetic efferents, cardiovascular responses, and experimental hypertension. Neurons within the ventrolateral medulla are integral to several reflex pathways. The activation of neurons within the A5 cell group can generate complex patterns of sympathetic and cardiovascular response. The type of respiratory modulation of a sympathetic neuron may be associated with its function. Opioid mechanisms within the central nervous system are important in cardiovascular responses to hemorrhage. The central actions of angiotensin II and atrial natriuretic peptide may blunt the baroreceptor reflex in spontaneously hypertensive rats.

In anaesthetized cats intracellular recordings in the nucleus tractus solitarius (NTS) have been used to identify neurones receiving inputs from the carotid sinus nerve (SN), the aortic nerve and hypothalamic defence area (HDA). In forty-four neurones activated by SN stimulation HDA activation evoked an IPSP in fourteen (32%) and an EPSP in eight (11%). Those neurones receiving an HDA-evoked IPSP, and tested for reflex inputs, responded only to baroreceptor activation. Neurones receiving HDA-evoked EPSPs responded only to chemoreceptor activation. In three neurones unaffected by SN stimulation, short latency inputs from HDA were observed, suggesting direct monosynaptic inputs.

The origin of cardiac vagal preganglionic motoneurones in the rat is still controversial and knowledge of the chemistry of synaptic inputs onto these neurones is limited. In this investigation vagal preganglionic motoneurones innervating the heart were identified by the retrograde transport of cholera toxin conjugated to horseradish peroxidase (CT-HRP) combined with the immunocytochemical localization of 5-hydroxytryptamine. Injection of CT-HRP into the myocardium resulted in the retrograde labelling of neurones primarily in the ventral regions of the nucleus ambiguus (75.1%). Labelled neurones were also distributed in a narrow band through the reticular formation extending between the dorsal motor nucleus of the vagus nerve and the nucleus ambiguus (17.3%) as well as in the dorsal motor nucleus itself (7.6%). A combination of retrograde labelling with immunocytochemistry for 5-hydroxytryptamine revealed that the neuronal perikarya and the dendrites of cardiac vagal motoneurones in the nucleus ambiguus were often ensheathed in 5-hydroxytryptamine-immunoreactive axonal boutons. Electron microscopic examination of this material confirmed that there were synaptic specializations between these boutons and the cardiac vagal motoneurones. The identification of 5-hydroxytryptamine-containing synaptic inputs to this population of vagal motoneurones provides further detail towards the understanding of the regulation of heart rate by the parasympathetic nervous system.

Recently we have shown that the density of [3H]5HT binding sites in the carotid body (CB) and within the nucleus of the tractus solitarius (NTS) of the cat is increased following unilateral chronic sectioning of the carotid sinus nerve (CSN) (Dashwood et al 1990). [125I]endothelin (ET) binding sites have also been identified in these regions, but these “receptors” are unaffected by CSN section (Spyer et al 1991). There is physiological evidence that both 5HT (Kirby and McQueen 1984) and ET (Spyer et al 1991) affect chemoreflex activity in the cat. Here we report the effect of chronic chemoreceptor stimulation, by almitrine bismesylate, on the density of central and peripheral 5HT and ET receptors in the cat.

1. Intracellular and whole-cell patch recordings were made from sixty-seven neurones located in the nucleus tractus solitarii (NTS) in transverse slices of rat brainstem. 2. Baclofen at concentrations of 2-20 microM caused hyperpolarization from normal resting membrane potentials (Vm). This response was associated with a decrease in input resistance (Rm) tested by current pulses in discontinuous current clamp mode when membrane potential was restored to control level by current injection. In single electrode discontinuous voltage clamp mode, baclofen at these concentrations caused a small (< 50 pA) outward current associated with increased membrane conductance measured by voltage steps from holding potentials (Vh) of -50 or -60 mV. Current-voltage relations at these Vhs and the results of varying Vh between -50 and -110 mV during responses to baclofen gave a reversal potential of -73 mV. The amplitudes of baclofen responses were related to K+ concentration tested by comparing responses in media containing 1-24 mM extracellular K+, indicating that postsynaptically baclofen acts via a K+ conductance. 3. These effects were still apparent in the presence of tetrodotoxin (which did not abolish all spontaneous synaptic activity) and also in medium containing a combination of Co2+, the excitatory amino acid antagonist 6-cyano-7-nitro-quinoxaline-2,3-dione (CNQX) and the GABAA antagonist bicuculline which blocked synaptic activity. 4. The amplitude and frequency of spontaneous postsynaptic potentials (spPSPs) and spontaneous postsynaptic currents (spPSCs) were reduced by baclofen at concentrations (1 microM or less) which had no effect on membrane potential or holding current in current or voltage clamp recordings respectively. 5. The amplitude of evoked excitatory (evEPSPs/evEPSCs) and inhibitory (evIPSPs/evIPSCs) synaptic events elicited by electrical stimulation in the vicinity of the tractus solitarius (TS) was reduced by low concentrations of baclofen (250 nM-1 microM) which did not produce discernible postsynaptic responses. 6. In order to examine the effects of baclofen on excitatory synaptic events without contamination with inhibitory events, stimulation of the TS was carried out in the presence of bicuculline. Conversely to investigate actions on purely inhibitory synaptic responses experiments were carried out with CNQX in the bathing solution. Inhibitory synaptic responses could still be evoked, presumably by stimulation of interneurones in the vicinity of the TS. IPSPs/IPSCs were more sensitive to baclofen than EPSPs/EPSCs. 7. The effects of baclofen on membrane potential or holding current and PSP/PSCs were antagonized by 2-hydroxysaclofen (400 microM) confirming that baclofen was acting at gamma-aminobutyric acid (GABA)B receptors.(ABSTRACT TRUNCATED AT 400 WORDS)

γ-Aminobutyric acid immunoreactive perikarya and boutons in the nucleus tractus solitarius of the cat were examined at both the light and electron microscopic level. Immunoreactive neurones were found predominantly in the parvocellular subdivision of the nucleus tractus solitarius and to a lesser degree in all the other subdivisions of the nucleus tractus solitarius and the dorsal vagal motonucleus. All the immunoreactive perikarya observed were similar in size and morphology. γ-Aminobutyric acid immunoreactive boutons were observed througout the nucleus tractus solitarius. However, in contrast to its high content of immunoreactive perikarya the parvocellular subdivision contained the lowest density of immunoreactive boutons. Ultrastructural examination of immunoreactive boutons in the different regions of the nucleus tractus solitarius revealed that they formed synaptic specializations, predominantly with dendritic shafts, all of which were of the symmetric type. This pattern of innervation was observed throughout the medial, commissural, ventrolateral and parvocellular subdivisions of the nucleus tractus solitarius.

Since the inhibitory neurotransmitter γ-aminobutyric acid (GABA) has been implicated in virtually all neuronal systems, it would be surprising if it were not also involved in the pathways responsible for maintaining and controlling blood pressure and heart rate. However, the exact role that it plays in cardiovascular control is still under intensive investigation and GABAergic inhibitory connections between cell groups are still being mapped anatomically and functionally. In addition, in recent years it has become apparent that GABA can act on at least two types of receptor (GABAA and GABAB), which are both capable of mediating pre- and postsynaptic inhibition but function via different ionic mechanisms. Agonists acting at the GABAA-receptor subtype cause inhibition by opening of Cl− channels whereas agonists at the GABAB-receptor subtype cause postsynaptic inhibition via a K+-conductance (Newberry and Nicoll, 1985) and presynaptic inhibition of transmitter release probably through a reduction in Ca2+-conduc-tance (see Huston et al., 1990).

1. The cardiovascular responses, together with the effects on medullary sympathoexcitatory (vasomotor) neurones of the rostral ventrolateral medulla, of area postrema stimulation have been studied in vivo. 2. Electrical (10 Hz) or chemical stimulation using microinjections of L-glutamate of the area postrema produced a vasodepressor response and an inhibition of the medullary sympathoexcitatory neurones in the nucleus reticularis rostroventrolateralis (RVL), while similar stimulation in the adjacent nucleus tractus solitarii (NTS) caused increases in arterial pressure. 3. Single-pulse stimulation of the area postrema revealed at least three influences on the activity of RVL vasomotor neurones, one being excitatory and two inhibitory. 4. The inhibitions evoked in the medullary vasomotor neurones on area postrema stimulation were blocked by ionophoretic application of bicuculline, a GABAA receptor antagonist, without altering the excitatory input to the same neurones. Bilateral microinjections of bicuculline into the RVL in regions where the vasomotor neurones had been identified totally eliminated the vasodepression due to area postrema stimulation. 5. These data support a role for the area postrema in cardiovascular control. It is concluded that the area postrema exerts its action on cardiovascular control in part via GABAergic inhibition of the 'vasomotor' neurones in the nucleus reticularis rostroventrolateralis.

The primary function of the respiratory system, the gills and/or lungs, is to exchange gases between the external environment and the internal milieu of the organism. The cardiovascular system assists this function by transporting blood gases — O2 from the gill/lung capillaries to the tissue capillaries and CO2 from the tissue capillaries to the gill/lung capillaries. These processes of gas exchange and gas transportation are adjusted to maintain homeostasis in varying physiological circumstances — this is the basic cardiorespiratory function.

We have demonstrated by autoradiography, displaceable binding for [125I]endothelin-1 ([125I]ET-1), [125I]endothelin-2 ([125I]ET-2), and [125I]endothelin-3 ([125I]ET-3) in the cat carotid bifurcation as well as in the nucleus of the tractus solitarius, where baroreceptor and chemoreceptor afferents from the carotid body and sinus terminate. There was also significant binding in the nodose and superior cervical ganglia. Barosensory and chemosensory discharge was recorded from filaments of the carotid sinus nerve in cats anesthetized with pentobarbitone. Intra-carotid injection of ET-1 or ET-3 (4-402 pmoles) caused transient dose-related depression of baroreceptor discharge without any immediate effects on systemic blood pressure (BP) or heart rate; there was a delayed biphasic effect on BP. ET-1 had little effect on chemosensory discharge during the first 15 s post-injection, but there was a delayed (45-90 s) dose-related increase in discharge. The effects of all three ETs were qualitatively similar, and ET enhanced chemoexcitation evoked by either acetylcholine or sodium cyanide. Our results show that (a) ET binding sites are located in the baroreceptor and chemoreceptor afferent pathways and (b) ETs can influence afferent activity of baroreceptors and chemoreceptors. Further studies are needed to determine the significance of these findings, particularly with regard to reflex control of the cardiovascular system.

1. The present study has assessed the importance of neurones within the nucleus tractus solitarius (NTS) in mediating the cardiovascular response evoked from the parabrachial nucleus (PBN) in the decerebrate rabbit. Microinjection techniques were employed so that the magnitude of the circulatory responses elicited from the PBN could be compared before, and after, kainic acid or bicuculline were microinjected into restricted regions of the NTS. 2. Electrical stimulation of the PBN (both medial and lateral regions) evoked variable changes in heart rate, a pressor response, vasoconstriction in the hindlimb and an increase in renal sympathetic nerve activity. Glutamate injected into these regions of the PBN elicited a similar pattern of response except that a tachycardia was observed consistently. 3. Both electrical and chemical stimulation of restricted regions of the NTS evoked bradycardia and a depressor response together with an increase in femoral vascular conductance and an inhibition of activity in the renal nerve. 4. Chemical lesions placed in these regions of the NTS by microinjecting kainic acid were found to attenuate both the heart rate and arterial blood pressure responses elicited from sites in the medial and lateral PBN using either electrical or chemical stimulation. Equivalent effects were produced on microinjecting the GABAa receptor antagonist bicuculline into the NTS. 5. These data indicate that NTS neurones play a part in mediating the cardiovascular responses that are evoked from the PBN and suggest that the action of the PBN at the level of the NTS is mediated via a GABAergic mechanism.

Whilst there is a growing understanding of the neural organization of respiratory (Spyer et al., personal communication) and cardiovascular reflexes (Spyer, 1988), little is knovm concerning the pharmacology of these pathways. Indeed, there is as yet no convincing evidence to specify the transmitters released by the afferents innervating the arterial chemo- receptors and baroreceptors. As the afferents arising from these receptors terminate solely within the nucleus tractus solitarius (NTS), and this nucleus is rich with peptide-containing terminals and perikarya, it has been speculated that there might be some involvement of peptides in these reflexes (Heike et al., 1980).

There is both anatomical and neurophysiological evidence to indicate that the primary synapses in the arterial chemoreceptor reflex are located within the nucleus tractus solitarius (NTS). Indeed, it appears that these afferents make connections with neurones in a relatively restricted portion of the NTS, particularly the commissural nucleus and the medial subnucleus in close proximity to the obex (for review see Donoghue et al., 1984; Housley et al., 1987; Jordan and Spyer, 1987). In contrast, there is little detailed evidence to support the notion that the respiratory neurones of the ventrolateral NTS are contacted directly by these afferents. The role of intrinsic circuits within the NTS in mediating the chemoreceptor control of respiratory activity, and the attended influence of this control on cardiovascular activity, have thus become a focus of interest in our laboratory. This report will summarize the results of neurophysiological, neuroanatomical, and immunocytochemical studies that have been designed to address these questions.