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J F Paton

Publications and source records attributed to J F Paton.

At least 19 recordsLinked to original sources
PubMed2001

GABA(A) receptor epsilon-subunit may confer benzodiazepine insensitivity to the caudal aspect of the nucleus tractus solitarii of the rat.

1. Benzodiazepines (BZ) and barbiturates both potentiate chloride currents through GABA(A) receptors to enhance inhibition. However, unlike barbiturates BZ do not impair autonomic control of heart rate. We hypothesised that BZ might not significantly potentiate GABAergic transmission in the caudal nucleus of the solitary tract (cNTS), which is critically important for mediating the baroreceptor reflex. 2. In rat brain slices the BZ agonists chlordiazepoxide and midazolam (2 and 50 microM) did not significantly enhance currents evoked by GABA in voltage-clamped cNTS neurones. Chlordiazepoxide (50 microM) reversibly increased electrically evoked IPSPs in 5/10 rostral NTS (rNTS) neurones but only in 2/10 cNTS neurones. Pentobarbitone (50-100 microM) was effective in enhancing GABA(A)-mediated responses in all NTS neurones. An inverse BZ agonist, methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM; 1 or 10 microM), failed to depress GABA-induced currents in the cNTS. 3. Microinjections of midazolam (10 and 100 microM solutions) into the cNTS did not affect the baroreceptor reflex (P > 0.2) while pentobarbitone (100 microM) significantly and reversibly depressed it (gain decrease to 53 +/- 11 % of control, P < 0.01). 4. Reverse transcriptase polymerase chain reaction revealed the presence of alpha(1), alpha(2), beta(2), beta(3) and gamma(2) GABA(A) receptor subunit mRNA in the cNTS. No alternatively spliced variants of the alpha(1)- and gamma(2)-subunits were revealed. Moreover, GABA(A) epsilon-unit mRNA was found in both the cNTS and rNTS as two alternatively spliced transcripts. 5. Immunocytochemical analysis revealed numerous GABA(A) epsilon-subunit-positive neurones within the cNTS with significantly fewer epsilon-subunit-positive cells in the rNTS. 6. As incorporation of the epsilon-subunit in recombinant GABA(A) receptors may confer BZ insensitivity we propose that the paucity of BZ actions in the cNTS is due to a high level of epsilon-subunit expression. This is the first demonstration of a possible physiological impact of the epsilon-subunit on native GABA(A) receptors.

Animals↗
PubMed2001

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.

Animals↗
PubMed2001

Adenoviral vector demonstrates that angiotensin II-induced depression of the cardiac baroreflex is mediated by endothelial nitric oxide synthase in the nucleus tractus solitarii of the rat.

Angiotensin II (ANGII) acting on ANGII type 1 (AT1) receptors in the solitary tract nucleus (NTS) depresses the baroreflex. Since ANGII stimulates the release of nitric oxide (NO), we tested whether the ANGII-mediated depression of the baroreflex in the NTS depended on NO release. In a working heart-brainstem preparation (WHBP) of rat NTS microinjection of either ANGII (500 fmol) or a NO donor (diethylamine nonoate, 500 pmol) both depressed baroreflex gain by -56 and -67 %, respectively (P < 0.01). In contrast, whilst ANGII potentiated the peripheral chemoreflex, the NO donor was without effect. NTS microinjection of non-selective NO synthase (NOS) inhibitors (L-NAME; 50 pmol) or (L-NMMA; 200 pmol) prevented the ANGII-induced baroreflex attenuation (P > 0.1). In contrast, a neurone-specific NOS inhibitor, TRIM (50 pmol), was without effect. Using an adenoviral vector, a dominant negative mutant of endothelial NOS (TeNOS) was expressed bilaterally in the NTS. Expression of TeNOS affected neither baseline cardiovascular parameters nor baroreflex sensitivity. However, ANGII microinjected into the transfected region failed to affect the baroreflex.Immunostaining revealed that eNOS-positive neurones were more numerous than those labelled for AT1 receptors. Neurones double labelled for both AT1 receptors and eNOS comprised 23 +/- 5.4 % of the eNOS-positive cells and 57 +/- 9.2 % of the AT1 receptor-positive cells. Endothelial cells were also double labelled for eNOS and AT1 receptors. We suggest that ANGII activates eNOS located in either neurones and/or endothelial cells to release NO, which acts selectively to depress the baroreflex.

Adenoviridae↗
PubMed2001

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.

Anesthetics↗
PubMed2001

Reorganisation of respiratory network activity after loss of glycinergic inhibition.

gamma-Aminobutyric acid (GABA)-ergic and glycinergic inhibition is believed to play a major role in the respiratory network. In the present study we tested whether specific blockade of glycinergic inhibition resulted in changes in respiratory network interaction and function. Using the working heart-brainstem preparation from adult mice, we recorded phrenic nerve activity and the activity of different types of respiratory neurones located in the ventrolateral medulla. Strychnine (0.03-0.3 microM) was given systemically to block glycine receptors (Gly-R). During exposure to strychnine, post-inspiratory (PI) neurones shifted their onset of discharge into the inspiratory phase. As a consequence, the post-inspiratory phase failed and the rhythm changed from a three-phase cycle (inspiration, post-inspiration, expiration, with a frequency of about. 0.24 Hz) to a faster, two-phased cycle (inspiration expiration, frequency about 0.41 Hz). Inspiratory and expiratory neurones altered their augmenting membrane potential pattern to a rapidly peaking pattern. Smaller voltage oscillations at approximately 10 Hz and consisting of excitatory and inhibitory postsynaptic potential sequences occurred during the expiratory interval. Due to their high frequency and low amplitude, such oscillations would be inadequate for lung ventilation. We conclude that, under physiological conditions, glycinergic inhibition does indeed play a major role in the generation of a normal respiratory rhythm in adult mice. After failure of glycinergic inhibition a faster respiratory rhythm seems to operate through reciprocal GABAergic inhibition between inspiratory and expiratory neurones, while phase switching is organised by activation of intrinsic membrane properties.

Animals↗
PubMed2001

Baroreflex inhibition of cardiac sympathetic outflow is attenuated by angiotensin II in the nucleus of the solitary tract.

Homeostatic regulation of arterial pressure is maintained by arterial baroreceptors. Activation of these receptors results in an inhibition of sympathetic activity to the heart. It is known that angiotensin II in the nucleus tractus solitarii attenuates the baroreceptor reflex-evoked vagal bradycardia. Here, we determined whether the cardiac sympathetic component of the baroreceptor reflex could be modulated by angiotensin II in the nucleus of the solitary tract. An in situ, arterially perfused working heart--brainstem preparation of rat was employed and the sympathetic inferior cardiac nerve recorded. Increases in perfusion pressure caused a reflex bradycardia and inhibition of inferior cardiac nerve activity. Microinjection of angiotensin II (500 fmol) in the nucleus of the solitary tract attenuated significantly both the reflex bradycardia and inhibition of inferior cardiac nerve activity (P<0.01). The latter was reversible and sensitive to losartan, an angiotensin II type 1 receptor antagonist. In contrast, the peripheral chemoreceptor reflex evoked an increase in inferior cardiac nerve activity that was not affected by angiotensin II applied exogenously in the nucleus of the solitary tract. We conclude that within the nucleus of the solitary tract angiotensin II exerts a powerful and specific inhibitory modulation of the baroreceptor reflex control of sympathetic nerve activity destined for the heart. We suggest that our data may have clinical implications relating to hypertension, a condition when angiotensin II activity is heightened in the brain and the efficacy of the baroreflex is reduced.

Angiotensin II↗
PubMed2001

Properties of solitary tract neurones responding to peripheral arterial chemoreceptors.

Despite the highly integrated pattern of response evoked by peripheral chemoreceptor stimulation, limited information exists regarding the neurones within the nucleus of the solitary tract that mediate this reflex. Using a working heart-brainstem preparation, we describe evoked synaptic response patterns, some intrinsic membrane properties, location, morphology and axonal projections of physiologically characterised 'chemoreceptive' neurones located in the solitary tract nucleus in the rat. From 172 whole cell recordings, 56 neurones were identified as chemoreceptive since they responded to aortic injections of low doses of sodium cyanide (2-5 microg). Chemoreceptive neurones had a mean resting membrane potential of -52+/-1 mV and input resistance was 297+/-15 M(Omega) (n=56). Synaptic responses evoked included excitatory synaptic potentials alone, excitatory-inhibitory post-synaptic potential complexes, inhibitory synaptic potentials alone and central respiratory modulated synaptic potentials. Synaptic response latency data were obtained by stimulating electrically the solitary tract: the mean excitatory synaptic latency was 5.2+/-0.4 ms (range 2.5-8.0 ms; n=17). Chemoreceptive neurones showed a heterogeneity in their intrinsic membrane properties: neurones displayed either steady state, augmenting or adapting firing responses to depolarising current injection and, in some neurones, either delayed excitation or rebound activity following hyperpolarising pulses. Eleven chemoreceptive neurones were labelled and provided the first morphological data of these cells. Labelled somata were detected dorsomedial or medial to the solitary tract spanning the obex. Neurones typically had three to eight primary dendrites which often entered the solitary tract as well as extending across the ipsilateral region of the nucleus of the solitary tract. Axons were mostly unmyelinated with boutons of the en passant variety and often ramified within the solitary tract nucleus as well as coursed towards the ipsilateral ventral medulla. In summary, this study provides new data on the neurophysiological, anatomical and morphological properties of nucleus of the solitary tract neurones responding to arterial chemoreceptors in the rat.

Action Potentials↗
PubMed2001

Unravelling mechanisms of action of angiotensin II on cardiorespiratory function using in vivo gene transfer.

We review recent and ongoing work from our laboratory that has shed novel insights into the effects of angiotensin II (ANGII) on the baroreflex at the level of the nucleus of the solitary tract (NTS). The NTS is the site of termination for baroreceptor afferents and is a potentially powerful region for neuronal modulation. ANGII applied to this nucleus attenuated the cardiac vagal and cardiac sympathetic components of the baroreceptor reflex. This effect was antagonized by blockade of either gamma-amino butyric acid receptors or nitric oxide synthase within the NTS. Interestingly, nitric oxide donors microinjected into the NTS mimicked the effect of ANGII. Using an adenovirus we showed that ANGII activated the endothelial isoform of nitric oxide synthase. The NTS was transfected to express a dominant negative truncated mutant form of endothelial nitric oxide synthase that prevented the depressant effect of ANGII on the baroreflex. Endothelial nitric oxide synthase was present in both neurones and endothelium in the NTS. A possibility is that ANGII activation of endothelial nitric oxide synthase is calcium dependent. However, in most NTS neurones tested, ANGII failed to elevate intracellular calcium concentration. We conclude that ANGII activates endothelial nitric oxide synthase to release nitric oxide which enhances gamma-amino butyric acid transmission destined for circuitry mediating the baroreflex. We discuss the contribution of endothelial cells within the nucleus of the solitary tract as a potential target for both circulating and/or centrally produced ANGII. These data have relevance to patients with essential hypertension and left heart failure, conditions in which ANGII activity is elevated and the baroreceptor reflex is depressed.

Adenoviridae↗
PubMed2001

Response properties of baroreceptive NTS neurons.

Neurons in the nucleus of the solitary tract (NTS) responding to activation of arterial baroreceptors were recorded intracellularly using patch pipettes in an in situ arterially perfused working heart-brain stem preparation of rat. Seven of 15 (i.e., 46%) of NTS neurons showed adaptive (nonlinear) excitatory synaptic response patterns during baroreceptor stimulation followed by an "evoked hyperpolarization." This evoked hyperpolarization was stimulus intensity dependent and capable of shunting out a subsequent baroreceptor input. We suggest that this adaptive response behavior may be mediated, in part, by calcium-dependent potassium currents (IKCa) since neurons showed spike frequency adaptation during step depolarizations and an after-hyperpolarization after repetitive firing. Furthermore, in in vivo anesthetized rats, NTS microinjections of either charybdotoxin (225 fmol) or apamin (4.5 pmol) to block IKCa increased the baroreceptor reflex gain. Our data purport that the responsiveness of baroreceptive NTS neurons can be regulated by intrinsic membrane conductances such as IKCa. Modulation of such conductances during either physiological (exercise) or pathophysiological (essential hypertension) conditions may lead to changes in both the operating point and gain of the baroreceptor reflex.

Adaptation, Physiological↗
PubMed2001

Brain stem PO(2) and pH of the working heart-brain stem preparation during vascular perfusion with aqueous medium.

The rat working heart-brain stem preparation (WHBP) is an in situ preparation having many of the advantages associated with in vitro preparations while retaining cardiovascular response functionality and an eupnoeic respiratory motor pattern. The preparation is perfused arterially with an aqueous medium having a much lower oxygen-carrying capacity than blood. To evaluate the efficacy of the artificial perfusion in providing adequate gas exchange within the brain stem, we used polarographic PO(2) and pH microelectrodes to determine the tissue PO(2) and pH of the medulla oblongata at various depths. When the perfusate was equilibrated with 5% CO(2) and 95% O(2), average tissue PO(2) was 294 Torr and no hypoxic areas were encountered. Tissue pH was remarkably uniform throughout the tissue, and on average was only 0.04 +/- 0.02 pH units more acidic than that of the perfusate. Increasing the PCO(2) of the perfusate increased tissue PO(2) and decreased arterial resistance. Decreasing perfusate PCO(2) (while keeping pH constant) decreased tissue PO(2) and reduced the respiratory activity. These results suggest that arterial PCO(2), independent of arterial pH, is an essential variable in determining both respiratory drive and cerebrovascular perfusion. We conclude that the medulla of the WHBP is oxygenated and within a physiological pH, which accounts for the eupneic pattern of respiratory motor activity it generates. Furthermore, this preparation may be a useful model for exploring mechanisms of central chemoreception as well as the dynamics of the cerebral vasculature responses following changes in blood gases.

Animals↗
PubMed2000

Respiratory activity in neonatal rats.

In neonatal animals in vitro preparations have been employed widely to study the central control of respiration. These preparations have limitations in that reflex afferent inputs and kinesiological studies cannot be performed. Here, we describe an alternative in situ experimental model for studying both peripheral and central control of the respiratory system in neonatal rats. Using technology based on adult mammals, we introduce an intra-arterially perfused working heart-brainstem preparation (WHBP) that permits studies on eupnoeic respiration in neonatal rats from within a few hours of birth. Using this preparation we demonstrate a three-phase respiratory rhythm as revealed by the activity in phrenic and recurrent laryngeal motor nerves, the respiratory modulation of laryngeal resistance and the firing patterns of respiratory neurones recorded from the ventrolateral medulla. We conclude that the neonatal rat WHBP is an in situ preparation because it produces a respiratory rhythm similar to that of adult in vivo mammal preparations but distinct from in vitro preparations.

Animals↗
PubMed2000

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.

Afferent Pathways↗
PubMed2000

Glutamate stimulation of raphe pallidus attenuates the cardiopulmonary reflex in anaesthetised rats.

Serotonin has been implicated as having a modulatory action on NTS neurones mediating vagal inputs controlling the cardiorespiratory system. Since raphe pallidus and raphe obscurus both send serotonergic projections to the NTS, we have investigated a neuromodulatory role for these structures on the cardiopulmonary reflex. A multibarrelled microelectrode positioned around the level of the area postrema was placed at varying depths into mid-line brainstem structures and the effect of glutamate stimulation on the cardiopulmonary reflex tested. Excitatory chemical stimulation in the region of raphe pallidus, but not raphe obscurus, attenuated significantly the respiratory and bradycardic components of the cardiopulmonary reflex. This attenuation was reversed by an NTS microinjection of RS-39604, a selective 5-HT, receptor antagonist. We propose that neurones in raphe pallidus that project to the NTS can release serotonin which acts via 5-HT4 receptors to attenuate the reflex phrenic nerve activity and heart rate components of the cardiopulmonary reflex.

Anesthesia↗
PubMed2000

Morphological and electrophysiological properties of neurones in the dorsal vagal complex of the rat activated by arterial baroreceptors.

This study physiologically identifies and anatomically describes arterial baroreceptive neurones in the nucleus tractus solitarii of the rat. Neurones were recorded using neurobiotin-containing whole cell patch electrodes in a working heart-brainstem preparation and characterized physiologically as arterial baroreceptive in response to stimulation of the aortic arch and/or ipsilateral carotid sinus. Fifteen of 84 neurones tested were arterial baroreceptive, 7 of 8 were morphologically identified as located in the solitary tract nucleus (NTS), and 1 of 8 was located in the dorsal vagal nucleus. The seven NTS neurones had a resting membrane potential of -52 +/- 3.6 mV and a membrane input resistance of 233 +/- 38 M omega. Action potential height was 62 +/- 4.2 mV, width at half amplitude 1.46 +/- 0.38 ms, and duration of after-hyperpolarization 1.7 +/- 2.33 ms. In six of eight neurones labelled there was an invariant excitatory synaptic input (latency 3.95 +/- 0.3 ms) to stimulation of the solitary tract. Labelled somata were dorsomedial or medial to the solitary tract from -0.3 mm to +1.5 mm with regard to obex. Neurones had three to eight primary dendrites, and branches often entered the solitary tract and also extended across the ipsilateral NTS. Axons, which were mostly unmyelinated with boutons of the en passant variety, could ramify within the NTS while the main axon exited the NTS (n = 4/6), in the direction of the ipsilateral ventral medulla (n = 5/6). This is the first morphological and localisation data of physiologically characterised arterial baroreceptive NTS neurones in the rat. By comparing labelled cells with each other as well as with other unidentified cells, we conclude that NTS arterially baroreceptive neurones are morphologically and physiologically heterogenous.

Animals↗