Differential increases in P2X receptor levels in rat vagal efferent neurones following a vagal nerve section

Extracellular ATP can influence cells via activation of P2X purinoceptors, the distribution of which can be altered in the central and peripheral nervous systems following injury or tissue damage. Here we have investigated the effect of a unilateral section of the cervical vagus nerve on the distribution of P2X(1), P2X(2), P2X(3), P2X(4) and P2X(7) receptor subunit immunoreactivity (R-IR) in the dorsal vagal motor nucleus (DVN) and the nucleus ambiguus (NA) in the medulla oblongata. As early as 2 days, and followed up to 14 days, there was a dramatic ipsilateral increase in P2X(1), P2X(2) and P2X(4)R-IR in the cell soma of vagal efferent neurones in the DVN following the nerve section, but not the NA. There were no changes in P2X(3) and P2X(7)R-IR in either nuclei. To test for possible functional consequences of increased P2X receptor levels, whole-cell patch-clamp recordings were made from DVN cells in brainstem slices 4 days following unilateral vagotomy. Application of ATP revealed large cell-to-cell variance in the current amplitude in neurones from both sectioned and control DVN. However, when ATP responses were compared to those elicited by the nicotinic acetylcholine receptor agonist carbachol, the mean ratio of the peak ATP-evoked current to the peak carbachol-evoked current was significantly larger in DVN neurones ipsilateral to the section. Thus the increase in P2XR levels in DVN cells ipsilateral to a nerve section are likely to reflect an increase in expression of functional P2XRs on the cell surface.     

A GABAergic mechanism in the inhibition of cardiac vagal reflexes

A depressor response and bradycardia were produced by aortic nerve stimulation in urethane-chloralose-anaesthetized, paralyzed and artificially ventilated rabbits. Stimulation of the superficial peroneal nerve (SP) or rostral ventrolateral medulla (rVLM) inhibited the aortic nerve stimulation-evoked bradycardia, which was also inhibited by direct microinjection of GABA into the dorsal vagus nucleus (DVN). Application of bicuculline methiodide into the same medullary area antagonized the effect of GABA and partially or completely abolished the SP or rVLM stimulation-produced inhibition. However, strychnine hydrochloride had no effect on the SP or rVLM stimulation-produced inhibition. These observations indicate that GABAergic system present at the DVN is possibly involved in the inhibition of evoked bradycardia during somatic afferent and rVLM stimulation.     

Stimulation of the ventrolateral medulla inhibits the baroreceptor input to the nucleus tractus solitarius

Extracellular recording experiments were done in urethane- and chloralose-anaesthetized, paralyzed and artificially ventilated rabbits. Thirty-one baroreceptor-sensitive neurones were identified in the region of the nucleus tractus solitarius (NTS) by their excitatory responses to stimulation of the ipsilateral aortic nerve. A conditioning stimulus delivered to the ipsilateral or contralateral rostral ventrolateral medulla (rVLM) inhibited the excitatory responses of 80.0% (19 out of 24) neurones to an aortic test stimulus as early as 3 ms and extending as long as 400 ms after conditioning. The same inhibitory effect was also observed by application of excitant amino acid DL-homocysteic acid (0.2 M, 100 nl) into the ipsi- or contralateral rVLM area in 6 units. In 5 units inhibited by rVLM stimulation, evoked discharges were inhibited by prolonged electrical stimulation of the superficial peroneal nerve (SP) with low intensity and low frequency (0.1-0.3 mA and 5-10 Hz). These results provide the electrophysiological evidence for the suppressing effect of the rVLM on the excitatory responses of NTS neurones to baroreceptor afferent stimulation.     

Electrophysiological properties of paraventriculo-spinal neurones in the rat

In rats anaesthetized with urethane, electrical stimulation of the thoracic cord at T9-10 evoked antidromic response in neurones in the parvocellular portion of the ipsilateral paraventricular nucleus. Estimated conduction velocities in the spinally-projecting axons ranged from 0.6-5.9 m/sec. The majority (97%) of spinally projecting neurones were quiescent. Increases or decreases in the level of baroreceptor stimulation produced by intravenous injection of phenylephrine (1-5 micrograms) or sodium nitroprusside (10-15 micrograms) inhibited and excited amino acid-induced activity in 5/8 and 4/11 neurones respectively. Stimulation of vagal afferent endings by rapid bolus injection of 5-HT, to evoke the Bezold-Jarisch reflex. inhibited amino acid-induced activity in 15/18 cells. Activation of gastric vagal afferents by distension of the stomach had no effect on paraventriculo-spinal cells. It is suggested that at rest the excitability of paraventriculo-spinal neurones is depressed by a tonic inhibitory input which arises from vagal afferent fibres in the heart.     

Convergent inputs to neurones in the nucleus paragigantocellularis lateralis in the cat

Extracellular recordings were made in the nucleus paragigantocellularis (PGL) of the cat in response to stimulation of the hypothalamic perifornical defence area (HPDA), dorsal periaqueductal grey matter (PAG), nucleus raphe obscurus (Rob), deep peroneal nerve (DPN), and superficial peroneal nerve (SPN). Stimulation of the HPDA, dorsal PAG and SPN evoked excitatory responses whilst the prevalent response to stimulation of Rob was inhibition. However, most of the defence-reaction-related neurones showed little response to stimulation of DPN. Of 53 cells tested 48 (91%) received convergent inputs from two or more sites of stimulation. These findings are discussed in relation to the integrative function of PGL in cardiovascular control.     

Responses of cells in the rat supraoptic nucleus in vivo to stimulation of afferent pathways are different at different times of the light/dark cycle

To determine whether the daily rhythms of spike activity in the supraoptic nucleus (SON) were accompanied by changes in the behaviour of its inputs, we used conventional extracellular single cell recordings from cells in the SON of anaesthetized rats while stimulating the contralateral optic nerve and the ipsilateral suprachiasmatic nucleus (SCN). Neurones in the SON region were identified by antidromic activation and classified as oxytocin or vasopressin cells, on the basis of their spontaneous firing patterns. Approximately 27% of both oxytocin (29/108) and vasopressin (39/147) neurones were excited by stimulation of the optic nerve, and the majority of responses had a long latency (>20 ms). Very few oxytocin (3/108) and vasopressin cells (2/147) were inhibited by stimulation of the optic nerve. The pattern of the responses (excitatory, inhibitory or nonresponsive) of oxytocin and vasopressin cells to stimulation of the optic nerve was significantly related to the time of day (chi-square test; P = 0.012, oxytocin cells; P = 0.006, vasopressin cells). The proportion of oxytocin cells excited by stimulation of the optic nerve was highest at ZT 4-8 and lowest at ZT 20-24. For vasopressin cells, it was highest at ZT 12-16 and lowest at ZT 20-24. The proportion of excitatory, inhibitory and complex responses seen in oxytocin and vasopressin cells following stimulation of the SCN also changed and was significantly different at different times of day (oxytocin cells: highest proportion of excitatory responses at ZT 12-16, P = 0.029; chi-square test; vasopressin cells: highest proportion of excitatory responses at ZT 0-4, P = 0.005; chi-square test). Thus, inputs to oxytocin and vasopressin neurones from the optic nerve and some outputs from the SCN changed during the light/dark cycle. Such changes may contribute to the generation of 24-h rhythms in activity of oxytocin and vasopressin neurones and release of the peptides.     

Target site of inhibition of baroreflex vagal bradycardia by nasal stimulation

We have previously reported that stimulation of nasal mucosa inhibits baroreflex vagal bradycardia (BVB) and this inhibition was mediated exclusively by the trigeminal nerve, and occurred principally at pontomedullary level. In this study, to identify the target site of the inhibition, several types of experiments were conducted in chloralose-urethane-anesthetized, beta-adrenergic receptor-blocked rats. Afferent discharges in the ethmoidal nerve (EN5) were increased in response to nasal stimulation by smoke, and electrical stimulation of the EN5 suppressed BVB induced by electrical stimulation of the aortic depressor nerve (ADN). Electrical stimulation of the EN5 inhibited vagal bradycardia evoked by either electrical or chemical stimulation of the nucleus tractus solitarius (NTS), while it rather facilitated bradycardia by stimulation of the nucleus ambiguus (NA) region. Microstimulation of the NTS induced antidromic compound spike potential along the ADN but this was not affected by stimulation of the EN5. ADN-evoked field potentials and unitary responses of neurons in the NTS were suppressed by stimulation of the EN5. These results suggested that barosensitive neurons in the NTS are the major target sites of inhibition of BVB by nasal stimulation in rats.     

Visceral afferent stimulation-evoked changes in the release of peptides into the parabrachial nucleus in vivo

Previous investigations have demonstrated that the peptides substance P (SP), calcitonin gene-related peptide (CGRP), cholecystokinin (CCK), neurotensin (NT) and somatostatin (SOM) significantly modulate the glutamate-mediated transmission of visceral information through the parabrachial nucleus (PBN) to the ventrobasal thalamus. In addition, we have shown that the staining intensity of SOM, CCK and NT in the PBN decreases significantly following 2 h of vagal stimulation as visualized using immunohistochemistry. As well, the staining intensity of both SP and CGRP in the PBN were shown to increase under similar conditions. The present investigation was done to determine whether the altered peptide staining intensity of these peptides observed following 2 h of vagal stimulation was the result of an altered peptide release from terminals within the PBN. Male Sprague-Dawley rats were anesthetized with sodium thiobutabarbitol and instrumented to record blood pressure and heart rate and for the stimulation of the cervical vagus nerve. A push–pull perfusion cannula was lowered into the region of the PBN for the continuous sampling of extracellular fluid. Radioenzymatic quantification of the perfusates for peptide content revealed that the extracellular fluid concentration of CGRP and SP increased significantly during the 2 h of vagal stimulation. When the vagal stimulation was terminated, the release of both CGRP and SP decreased significantly below prestimulated values for approximately 30 min before returning to prestimulated levels shortly thereafter. In contrast, there was a significant decrease in the release of CCK, SOM and NT into the PBN during the period of vagal stimulation. Extracellular perfusate levels of these peptides returned to normal upon termination of stimulation. These results demonstrate that terminal release of CGRP and SP is significantly increased and terminal release of CCK, SOM and NT is significantly decreased in the PBN during 2 h of vagal stimulation. These results are consistent with our previous finding that the immunohistochemical staining intensity of CGRP and SP is increased while that of CCK, SOM and NT is decreased following vagal stimulation.     

Electrophysiological evidence for axonal branching of ambiguous laryngeal motoneurons

Laryngeal motoneurons in the nucleus ambiguous (NA) were identified antidromically by stimulation of the ipsilateral superior laryngeal nerve (S) and/or the recurrent laryngeal nerve (R). In some NA motoneurons, antidromic spikes elicited by both S and R stimulation collided with the spontaneously occurring discharges. In the same neuron, spikes evoked antidromically by stimulation of one laryngeal nerve always collided with antidromic spikes elicited by stimulation of the other laryngeal nerve. Of 105 NA neurons activated by S and R stimulation, 36 neurons satisfied the criteria, and were classified as NA neurons with branching axons (branching NA (B-NA) neurons). Those neurons activated by either S or R stimulation but not both were classified as NA neurons without branching axons (unbranched NA (UB-NA) neurons). Mean antidromic latencies of B-NA neurons were 0.79 +/- 0.20 ms to S stimulation and 1.91 +/- 0.45 ms to R stimulation and those values for UB-NA neurons were 0.84 +/- 0.17 ms to S stimulation and 2.10 +/- 0.53 ms to R stimulation respectively. None of these mean values were significantly different from one another. Conduction time in the unbranched portion of the branching axon was estimated according to the equation reported by Anderson and Yoshida. The mean conduction time for 20 B-NA neurons was 0.45 +/- 0.35 ms. The branching point in B-NA neurons was estimated on the basis of the conduction time in the unbranched stem portion and those times in two branches of a branching axon measured electrophysiologically. The results suggest that the majority of B-NA neurons bifurcate within a half axonal length.(ABSTRACT TRUNCATED AT 250 WORDS)     

Localization of cardiac vagal preganglionic motoneurones in the rat: Immunocytochemical evidence of synaptic inputs containing 5-hydroxytryptamine

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-hydroxytrypta-mine 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. © 1993 Wiley-Liss, Inc.     

A study of the substance P innervation of the intermediate zone of the thoracolumbar spinal cord

Immunocytochemical procedures have been used to examine the distributions of substance P (SP)-positive fibres within the intermediate zone of the thoracolumbar spinal cords of rabbits, cats, and monkeys. In all three species SP fibres were concentrated in areas known to contain sympathetic preganglionic neurones. These included the intermediolateral nucleus and the funiculus just lateral to it, the medial gray matter in the area of the nucleus intercalatus, and the paracentral region. The density of the SP innervation varied in a characteristic way both between these subpopulations of sympathetic neurones and in its overall input to different segmental levels. Generally the greatest accumulations of SP fibres were found in the T3-T5 and L2-L4 regions and these were concentrated in the intermediolateral nucleus (ILN). The highest densities of SP fibres in the lateral funiculus were in the upper thoracic and upper lumbar segments whereas SP fibres forming transverse bands, possibly in association with neurones in the nucleus intercalatus, were most prominent in T5-T8. Substance P fibres adjacent to the midline were more or less equally dense throughout the segments examined. Substance P-positive cell bodies situated immediately lateral to the central canal were present at a density of 200-300 per segment throughout the cat thoracolumbar cord. These neurones may be the cells of origin of at least some of the SP fibres in the intermediate zone. The close association of sympathetic preganglionic neurones with SP fibres, many of which are thought to be derived from cells in the medulla, suggests a role for SP-containing fibres in the modulation of sympathetic activity. The variation in input to different segments and classes of sympathetic neurones further suggests a specificity which may be related to the different functions of the neurones innervated.     

Neurones in the chicken ureter are innervated by substance P- and calcitonin gene-related peptide-containing nerve fibres: Immunohistochemical and electrophysiological evidence

Numerous ganglia or single neurones immunoreactive to protein gene-product 9.5 (PGP) were demonstrated in the chicken ureter. Ganglia were observed in the main nerve trunks accompanying the ureter (400–2,000 cells), in the adventitia (1–45 cells; density; 79 ± 12 ganglia/cm²; mean ± S.E.M.), in the circular muscle (1–9 cells; 76 ± 10 ganglia/cm²) and in the longitudinal muscle (1–8 cells; 232 ± 41 ganglia/cm²). Most of the PGP-positive neurones in the nerve trunk ganglia (∼66%) and in the smooth muscle layers (85%) were encircled by a dense plexus of varicose nerve fibres containing both substance P (SP) and calcitonin gene-related peptide (CGRP). SP-positive somata were rarely observed. Immunogold electron microscopy revealed that SP- and CGRP-immunoreactivity were colocalised in the same dense core vesicles. A strong reduction of SP-positive nerve fibres was observed in organ cultures of the ureter, indicating their extrinsic origin. The fibres might originate from the dorsal root ganglia, where SP and CGRP were colocalised in 20–30% of the neurones. The sensitivity of ureteric neurones to SP and CGRP was investigated in recordings obtained from mechanosensitive nerve fibres with cell bodies located in or adjacent to the ureter (U-G units). The majority (71%) of the U-G units was excited by local application of SP in a dose-dependent manner. The SP-sensitive U-G neurones had higher mechanical thresholds (29 ± 5 mmHg) as opposed to the SP-insensitive ones (10 ± 3 mmHg). Repeated applications of high doses of SP to the U-G units resulted in desensitisation and reduced the response to mechanical stimuli. None of the U-G units responded to local application of CGRP, but all U-G units were excited by acetylcholine. The data support the hypothesis that SP-containing primary afferents are involved in the modulation of the activity of ureteric neurons in the chicken. J. Comp. Neurol. 380:105–118, 1997. © 1997 Wiley-Liss. Inc.     

Collaterals of recurrent laryngeal nerve fibres innervate the thymus: a fluorescent tracer and HRP investigation of efferent vagal neurons in the rat brainstem

The origin and course of efferent vagal fibers, which innervate the rat thymus, were investigated by a fluorescent retrograde double labeling method, using Fast blue (FB) and Diamidino yellow dihydrochloride (DY) as tracers. In the same animal, one tracer was injected into the cranial portion of the right lobe of the thymus and the other dye was deposited around the cut end of the right recurrent laryngeal nerve. The neuronal population giving origin to the recurrent nerve was mapped by using retrograde labeling with HRP applied to the central stump of the nerve. The HRP retrograde axonal transport showed that most efferent vagal fibers of the recurrent nerve have their perikarya in the nucleus retroambigualis (NRA), nucleus ambiguus (NA), and to a lesser extent in the nucleus retrofacialis (NRF). In fluorescent retrograde double labeling of thymus and recurrent laryngeal nerve both single and double labeled cells were found. The cells labeled by the injections into the thymus were colocalized with the neurons labeled by the tracer deposited in the recurrent laryngeal nerve to the NRA, NA, and NRF. Moreover along the rostrocaudal extent of the NRF and NA double labeled cells were present, showing that some of the thymic efferents are collaterals of the recurrent nerve fibers. Our experiments shown that some thymic vagal fibres originate from neurons of nucleus dorsalis nervi vagi (NDV) as demonstrated both by HRP and FB injected thymuses. The possible role of these efferents in thymic function is briefly discussed.     

Responses of pontocerebellar neurones to stimulation of the parietal association and the frontal motor cortices.

The corticopontine projections from the parietal association cortex (the anterior portion of the middle suprasylvian gyrus) were electrophysiologically investigated and compared with those from the frontal motor cortex (the anterior sigmoid gyrus) in cats under light Nembutal anaesthesia. It was indicated by field potential study that the pontine nucleus (PN) neurones receive a significant amount of the direct corticopontine fibres from both the parietal and frontal cortical areas. In extracellular unitary study, out of 107 PN neurones identified by antidromic activation due to the brachium pontis stimulation, 33 responded with firings to stimulation of the parietal association area and 64 to the frontal motor area. Only 10 of them were excited by both parietal and frontal stimulations, but they were not estimated to receive the dual monosynaptic projections from both cortical areas. There were found data suggesting that the pontocerebellar neurones with faster conduction velocities respond at shorter latencies to the cortical stimulation and those with slower conduction velocities fire at longer latencies on the cortical stimulation. No remarkable difference was observed between the topographical localization of the PN neurones receiving the projection fibres from the parietal association and the frontal motor cortical areas. It was concluded that a vast majority of the pontocerebellar neurones possibly receive monosynaptic contacts differentially with the corticopontine fibres originating from the parietal association and the frontal motor areas.     

Source and origin of nerve fibres immunoreactive for substance P and calcitonin gene-related peptide in the normal and chronically denervated superior cervical sympathetic ganglion of the rat

Immunohistochemical studies of sympathetic ganglia have indicated that the normal rat superior cervical ganglion contains both SP-IR and CGRP-IR fibres, and CGRP- and SP-immunoreactivity coexist in some fibres. In rat sympathetic ganglia decentralization by preganglionic denervation leads to intraganglionic increase of peptidergic fibres immunoreactive (IR) for substance P (SP) and calcitonin gene-related peptide. We explored the sources of SP- and CGRP-IR fibres in normal and in chronically decentralized rat SCGs.The distribution of immunoreactivities for CGRP and SP was determined in SCGs of normal rats and of rats following preganglionic denervation followed by sensory denervation. Ganglia were studied after short-term (2–5 days) sensory denervation, and long-term (7–16 months) sympathetic denervation followed by short-term (2 days) sensory denervation. To explore for the production of SP and CGRP by intrinsic neurones within the ganglion, normal and chronically decentralized SCGs were examined following pretreatment by local in vivo application of colchicine. Normal and chronically decentralized ganglia were also injected with fluorescent tracer Fluorogold for retrograde tracing of extrinsic fibres back to their neurones of origin.The observations suggest that in normal SCG in the rat the SP-IR and CGRP-IR nerve fibres are derived via direct links from vagus and glossopharyngeal nerves and the cervical plexus, or from nerve fibres running along the cervical sympathetic trunk, and the external carotid and the internal carotid nerves.Sensory nerve inputs to the rat SCG following decentralization may contribute to the low levels of ganglionic activation observable in the autonomic failure of multiple system atrophy in man.     

The role of 5-HT, GABA and opioid peptides in presynaptic inhibition of tooth pulp input from the medial brainstem

In decerebrate or chloralose-anaesthetized cats electrical stimulation in the spinal trigeminal nucleus evoked antidromic responses in the mandibular canine tooth pulp. Conditioning stimulation in nucleus raphe magnus (NRM) and in the adjacent contralateral medullary reticular formation, nucleus reticularis gigantocellularis (NRGC) and nucleus reticularis magnocellularis (NRMC), produced a decrease in the threshold for the antidromic responses in a proportion of the tooth pulp inputs. This was interpreted as being due to depolarization of the tooth pulp afferent terminals, reflecting presynaptic inhibition. The primary afferent depolarization (PAD) of tooth pulp afferent terminals by NRM stimulation could be selectively blocked by bicuculline applied intravenously or by iontophoresis in the terminal region. Intravenous naloxone, cinanserin and methysergide had no effect on the PAD evoked from NRM, NRGC or NRMC. Thus NRM appears to exert presynaptic inhibitory control of A delta tooth pulp input to the spinal trigeminal nucleus via GABA-containing neurones.     

Separate medullary pathways mediatiating reflex vagal bradycardia to stimulation of buffer nerves in the cat

In this investigation of medullary pathways mediating reflex vagal bradycardia to stimulation of buffer nerves, four series of experiments were done in 37 cats anesthetized with chloralose, paralyzed and artificially ventilated. In the first series using animals in which the contralateral vagus was sectioned and the spinal cord was transected at C2, simultaneous stimulation of the aortic depressor (ADN) and carotid sinus (CSN) nerves elicited a bradycardia of magnitude not significantly different from the algebraic sum of the responses elicited by stimulation of the two buffer nerves separately, suggesting the existence of two separate central pathways mediating vagal bradycardia. In the second series, in spinal unilaterally vagotomized animals, lesions of the nucleus ambiguus (AMB) selectively attenuated the ADN reflex vagal bradycardia but not the CSN response; on the other hand, lesions of the external cuneate nucleus (ECN) attenuated the reflex vagal bradycardia elicited by stimulation of the CSN, but did not alter the ADN response. In the third series of experiments, 153 spontaneously firing single units in the region of the AMB and 98 in the region of the ECN were tested for responses to ADN and CSN stimulation. In the AMB 42% of the responsive units were excited only by stimulation of the ADN, 25% were activated only by stimulation of the CSN and 33% were activated by stimulation of both buffer nerves. Of the single units in the ECN region 85% were excited only by stimulation of the CSN, 15% during stimulation of both the CSN and ADN and none responded to stimulation of only the ADN. In the final series, electrical stimulation of the ECN evoked antidromic compound action potentials in the CSN but not in the ADN. Electrical stimulation of the AMB did not evoke antidromic activity in either the CSN or ADN. These studies provide evidence for the existence of two separate medullary pathways mediating the vagal reflex bradycardia to stimulation of the ADN and CSN.     

Gating of vagal inputs by sciatic afferents in nonspinally projecting neurons in the rat rostral ventrolateral medulla oblongata

Integration and coordination of somato-visceral sensory information is crucial to achieve adaptive behavioural responses. We have recently shown that sensory vagal and somato-sensory (sciatic nerve) inputs converge in neurons of the rostral ventrolateral medulla oblongata, which was implicated in adjusting visceral activities to changing somatic performances. In the present study, the neuronal mechanism of interaction between sciatic and vagal sensory inputs was examined in the rostral ventrolateral medulla oblongata using in vivo intracellular recording and labelling. Conditioning stimulation of the contralateral sciatic nerve (2 V) led to a time-dependent inhibition of responses to vagal stimulation (100 microA) in each RVLM neuron that received convergent sciatic and vagal sensory inputs (n = 50). None of these neurons had direct spinal projections, and only 8% of them exhibited a visible response to stimulation of the aortic depressor nerve. A significant attenuation of the amplitude of vagal test responses was present for up to 800 ms of conditioning delay, although the duration of this sciatico-vagal inhibition was greatly dependent on the intensity of both stimuli. The electrophysiological data indicated that sciatico-vagal inhibition is mediated presynaptically, via activation of GABAB receptors. Morphological evidence of axo-axonic interactions that may underlie sciatico-vagal inhibition was subsequently found in the electron microscope. It is suggested that during movements of the hindleg, activation of sciatic sensory fibres leads to re-patterning of neuronal activity in RVLM neurons via inhibition of visceral sensory inputs. Sciatico-vagal inhibition is likely to affect the activity of those RVLM neurons that modulate higher neuronal activities via ascending projections.     

Sciatic and vagal sensory inputs converge onto non-baroreceptive neurones of the rostral ventrolateral medulla

Previous data suggested that somatic and vagal sensory afferent inputs may converge in the rostral ventrolateral medulla oblongata (RVLM). The aim of the present study was to establish the existence of convergence between inputs mediated via the cervical vagus and contralateral sciatic nerves using in vivo intracellular recordings. The majority of RVLM neurones that received input from the vagus or the sciatic nerves also responded to stimulation of the other nerve. In 72% of the neurones the response was excitation or inhibition to stimulation of both nerves, respectively. The most frequent response type was a short excitation in response to stimulation of both nerves. Only 8% of the neurones exhibited a visible response to stimulation of the aortic depressor nerve. The results provided experimental evidence that non-baroreceptive neurones of the RVLM are involved in coordination of somatic and visceral sensory inputs.     

Burst-firing activity of presumed 5-HT neurones of the rat dorsal raphe nucleus: electrophysiological analysis by antidromic stimulation

We recently reported raphe neurones which frequently fired spikes in short bursts. However, the action potentials were broad and the neurones fired in a slow and regular pattern, suggesting they were an unusual type of 5-hydroxytryptamine (5-HT) neurone. In the present study, we investigated whether these putative burst-firing 5-HT neurones project to the forebrain and whether all spikes fired in bursts propagate along the axon. In anaesthetised rats, electrical stimulation of the medial forebrain bundle evoked antidromic spikes in both burst-firing neurones and in single-spiking, classical 5-HT neurones recorded in the dorsal raphe nucleus. Although the antidromic spike latency of the single-spiking and burst-firing neurones showed a clear overlap, burst-firing neurones had a significantly shorter latency than single-spiking neurones. For both burst-firing neurones and classical 5-HT neurones, antidromic spikes made collisions with spontaneously occurring spikes. Furthermore, in all burst-firing neurones tested, first, second and third order spikes in a burst could be made to collide with an antidromic spike. Interestingly, in a small number of burst-firing neurones, antidromic stimulation evoked spike doublets, similar to those recorded spontaneously. From these data we conclude that burst-tiring neurones in the dorsal raphe nucleus project to the forebrain, and each spike generated by the burst propagates along the axon and could thereby release transmitter (5-HT).