The inhibitory effect of somatic inputs on the excitatory responses of vagal cardiomotor neurones to stimulation of the nucleus tractus solitarius in rabbits

Experiments were done in 41 rabbits anaesthetized with urethane and chloralose, paralyzed with Flaxedil and ventilated artificially. Extracellular recordings of 142 units were made in the dorsal vagal nucleus (DVN) and the nucleus ambiguus (NA), identified by antidromic response to stimulation of the cervical vagus nerve. In total 63.5% of them exhibited spontaneous activity and 22 units (17 in DVN and 5 in NA) showed a cardiac rhythm; their antidromic conduction velocity was 3.7-12.5 m/s, which suggests their having axons in the range of B fibres. These neurones were classified as vagal cardiomotor neurones. A total of 16 DVN and 4 NA vagal cardiomotor neurones were excited orthodromically by electrical stimulation of the contralateral nucleus tractus solitarius (NTS). Electrical stimulation of the superficial peroneal nerve (SP) with low intensity or the deep peroneal nerve (DP) with high intensity which activated C fibres inhibited excitatory responses of 16 neurones (14 in DVN and 2 in NA). The other 4 neurones were unaffected by SP inputs. These results provide electrophysiological evidence for the inhibitory effect of somatic inputs on the evoked discharges of vagal cardiomotor neurones in the DVN and the NA.     

Electron microscopic localisation of P2X4 receptor subunit immunoreactivity to pre- and post-synaptic neuronal elements and glial processes in the dorsal vagal complex of the rat

P2X receptors are ligand gated ion channels activated by extracellular ATP. There are seven P2X subunits, P2X(1-7), and all are expressed in the CNS. The P2X(4) receptor subunit (P2X(4)R) is likely to be important in the CNS as it has been reported to be expressed throughout the brain and spinal cord. However, P2X(4)Rs have been identified as restricted to neurones, only in glia or expressed in both neurones and glia with no discernible relationship to CNS region or epitope target of antibodies used for staining. In addition, although there are particularly high levels of mRNA encoding P2X(4)R in the brainstem, previous immunohistochemical studies have revealed only indistinct staining. We therefore examined the distribution of P2X(4)R in the dorsal vagal complex (DVC) of the brainstem using immunohistochemistry in sections obtained from adult Wistar rats transcardially perfused with aldehyde fixatives. When this revealed staining identifiable only as small puncta at the light microscope level, we examined the area with electron microscopy. This ultrastructural study revealed that P2X(4)R immunoreactivity (IR) was present in neurones at both pre- and post-synaptic sites as well as in glial cell processes and somata. This P2X(4)R-IR was localised adjacent to plasma membranes, as well as internally in membrane bound structures resembling endosomes. Immunoreactivity in endosomes was more prominent following antigen retrieval protocols. Localisation of P2X(4)R-IR in astrocytes, identified by the presence of glial fibrillary acidic protein (GFAP), was confirmed using immunofluorescence. The presence of P2X(4)Rs in the dorsal vagal complex is consistent with expression studies, but some reasons for a lack of correlation with pharmacological studies are discussed. The P2X(4)R is therefore expressed by neurones and glia in the dorsal vagal complex and may play a role in mediating extracellular signalling by ATP in this region.     

Reduced glutamate binding in rat dorsal vagal complex after nodose ganglionectomy

Quantitative receptor autoradiography with L-[3H]glutamate was employed to examine the distribution and properties of glutamate binding sites in the rat brain 14 days after excision of the right nodose ganglion. Slide-mounted coronal sections of the brain showed reduced L-[3H]glutamate binding in the nucleus tractus solitarius/dorsal motor nucleus of the vagus in the ipsilateral relative to the sham-operated side. Densitometric and saturation analyses of binding data indicated a significant reduction in the density of glutamate binding sites (57% decrease relative to sham), while there was a significant increase in receptor affinity (40% greater than sham). Binding was unaltered in the inferior olivary complex. Glutamate receptors are likely to exist on synaptic nerve terminals of vagal afferent fibres within the nucleus tractus solitarius and on vagal preganglionic neurones within the dorsal motor nucleus of the vagus and/or their dendritic processes within the nucleus tractus solitarius. Additionally, our receptor autoradiographic studies provide evidence for L-glutamate being a transmitter of vagal afferent neurones.     

Molecular analysis of the vagal motoneuronal degeneration after right vagotomy

The aim of this study was to investigate the vagal motoneuronal degeneration after right vagotomy using in situ hybridization, RT-PCR, and immunohistochemistry methods. The morphology of the vagal motoneurons in dorsal motor nucleus of the vagus nerve (DMV) and nucleus of ambiguus (NA) after right vagotomy was examined by using Nissl staing and TUNEL. The expression of inducible nitric oxide synthase (iNOS), bcl-2, bax, and caspase-3 in DMV and NA of rats after right vagotomy was studied. Additionally, the involvement of the N-methyl-D-aspartate (NMDA) receptor-calcium-neuronal nitric oxide synthase (nNOS) pathway in the vagal motoneuronal degeneration was addressed by double-immunolabeling analysis of nNOS with NMDAR1 and calbindin D28K in right-vagotomized rats. The neurons in right DMV and NA displayed a darkly stained, shrunken morphology at 1 day and 5 days following right vagotomy as shown by Nissl staining. Quantitative analysis revealed that, at 1 day and 5 days following right vagotomy, the number of neurons in right DMV, but not NA, was significantly reduced in comparison with that of control rats. Occasional TUNEL-positive neurons were detected in right DMV of rat at 1 day after right vagotomy. The expression of iNOS protein and mRNA was absent in DMV and NA of control rats. However, the iNOS mRNA expression was induced bilaterally in DMV and NA at 1 day postoperation and continued to be up-regulated until 5 days after vagotomy as shown by in situ hybridization. Immunohistochemistry analysis also showed the increased expression of iNOS in bilateral DMV and NA of vagotomized rats. RT-PCR analysis revealed the enhanced bcl-2 and reduced bax mRNA levels and subsequent up-regulation of both bcl-2 and bax mRNA in right sides of the vagotomized brainstems at 1 day and 5 days postoperation, respectively. In situ hybridization analysis confirmed the up-regulation of bcl-2 and bax mRNA in right DMV and NA of the rats at 5 days following operation. Immunohistochemistry analysis showed up-regulated Bcl-2 immunoreactivity and undetectable changes in Bax immunoreactivity in DMV and NA of rats at 1 day after vagotomy, whereas enhancement of both Bcl-2 and Bax immunoreactivity was observed at 5 days postoperation. In addition, the caspase-3 mRNA level was elevated ipsilaterally in DMV and NA at 1 day and 5 days following right vagotomy. Double-immunofluorescence analysis showed complete colocalization of nNOS with NMDAR1 and with calbindin in ipsilateral DMV and NA at 10 days following right vagotomy. This study suggests that the signal pathway for NMDAR1-calcium-nNOS and the up-regulation of iNOS in DMV and NA may be involved in the vagal motor neurodgeneration after right vagotomy. Furthermore, our results imply that the apoptosis pathway mediated by Bcl-2, Bax, and caspase-3 may be activated in vagal motoneurons after right vagotomy.     

Selective retrograde transport of 3H-serotonin in vagal afferents

A new serotonergic afferent vagal component has been demonstrated in the cat by radioautography. Twenty-four hours after a bilateral injection of tritiated serotonin (3H-5-HT) into the area of the nucleus of the solitary tract (NST), heavily and lightly labelled cell bodies were observed in the nodose ganglia. After unilateral injections of 3H-5-HT into the same area, labelled ganglionar cell bodies were found in the ipsilateral nodose ganglion. Some were also found in the contralateral one, suggesting a serotonergic crossed fibers component. Dense clusters of silver grains, depicting typical labelling of neuronal varicosities, were observed in the NST. After destruction of the serotonergic terminals with 5,7-dihydroxytryptamine, followed by injection of 3H-5-HT, the number of labelled cell bodies decreased dramatically in the ipsilateral nodose ganglia and the clusters of silver grains disappeared in the NST. After ligature or section of the supranodose vagal nerve, following injection of 3H-5-HT into the NST, no radioautographic reaction was observed in the homolateral nodose ganglia. The present study demonstrates the existence of a peripheral serotonergic system in vagal afferents. The physiological implications of this new serotonergic visceral pathway remain to be studied.     

Preprogalanin mRNA is increased in vagal motor neurons following axotomy.

Expression of preprogalanin and tyrosine hydroxylase mRNA was examined in the rat dorsal vagal complex following nodose ganglionectomy and cervical vagotomy, using in situ hybridization of specific 35S-labelled oligonucleotides. Seven days after unilateral cervical vagotomy (and nodose ganglionectomy), neurons in the ipsilateral dorsal motor nucleus of the vagus and nucleus ambiguus expressed 6- to 10-fold increased levels of preprogalanin mRNA. In contrast, tyrosine hydroxylase mRNA was no longer expressed by cells of the dorsal motor nucleus of the vagus after the lesion. These results demonstrate that changes in the expression of the galanin and tyrosine hydroxylase genes occur in vagal motor neurons following lesion of their axons. More generally, these results, and those from other laboratories, demonstrate that specific alterations of neuropeptide and neurotransmitter production, are part of the reactive process activated by nerve injury.     

Selective retrograde transport of H-serotonin in vagal afferents

A new serotonergic afferent vagal component has been demonstrated in the cat by radioautography. Twenty-four hours after a bilateral injection of tritiated serotonin (³H-5-HT) into the area of the nucleus of the solitary tract (NST), heavily and lightly labelled cell bodies were observed in the nodose ganglia. After unilateral injections of ³H-5-HT into the same area, labelled ganglionar cell bodies were found in the ipsilateral nodose ganglion. Some were also found in the contralateral one, suggesting a serotonergic crossed fibers component. Dense clusters of silver grains, depicting typical labelling of neuronal varicosities, were observed in the NST. After destruction of the serotonergic terminals with 5,7-dihydroxytryptamine, followed by injection of ³H-5-HT, the number of labelled cell bodies decreased dramatically in the ipsilateral nodose ganglia and the clusters of silver grains disappeared in the NST. After ligature or section of the supranodose vagal nerve, following injection of ³H-5-HT into the NST, no radioautographic reaction was observed in the homolateral nodose ganglia. The present study demonstrates the existence of a peripheral serotonergic system in vagal afferents. The physiological implications of this new serotonergic visceral pathway remain to be studied.     

FINE STRUCTURAL CHANGES IN NERVE CELL BODIES OF THE ADULT RABBIT DORSAL MOTOR VAGAL NUCLEUS DURING AXON REACTION

Counts of neuronal nucleoli were made in the dorsal motor vagal nucleus (DMV) of the adult rabbit 10, 18, 70 and 90 days following unilateral cervical vagotomy. The structural characteristics of nerve cell bodies in the DMV were studied electron microscopically 2–90 days after cervical vagotomy. The nucleolar counts indicated that 20% of the large DMV neurones had disappeared ipsilateral to the operation 10 days postoperatively (p. o.), 65% 18 days p. o. and 70% 70 and 90 days p. o. No loss of small neurones was found. Large neurones ipsilateral to the operation showed nuclear displacement, infoldings of the nuclear membrane and disappearance of granular endoplasmic reticulum beginning 4 days p. o. and being prominent 6–18 days p. o. At the peak of the response, 10–18 days p. o., reacting neurones showed nucleolar condensation and vacuolation, the appearance of intranuclear electron-dense particles, extensive accumulation of intracytoplasmic lipid droplets, increased numbers of microtubules and neurofilaments, focal mitochondrial aggregates, and widespread mitochondrial degeneration. Ten to 21 days p. o. degenerating neurones were observed. After 30 days p. o. survival a partial recovery of surviving large DMV neurones seemed to have taken place. The findings are interpreted as indications of distubed protein metabolism, oxidative metabolism and intraneuronal transport in the axotomized DMV neurones. The unique response of these neurones compared to previously studied peripherally projecting neurones is emphasized.     

Cross-inhibitory interactions between GABAA and P2X channels in myenteric neurones

Inhibitory interactions between GABA(A)[induced by gamma-aminobutyric acid (GABA)] and P2X [activated by adenosine 5'-triphosphate (ATP)] receptors of myenteric neurones from the guinea pig small intestine were characterized using whole-cell recordings. Currents induced by GABA (I(GABA)) or ATP (I(ATP)) were inhibited by picrotoxin or pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid, respectively. Currents induced by GABA + ATP (I(GABA+ATP)) were only as large as the current induced by the most effective transmitter, revealing current occlusion. This occlusion requires maximal activation of at least one of these receptors. Sequential applications of neurotransmitters, and kinetic and pharmacological properties of I(GABA+ATP) indicate that they are carried through both GABA(A) and P2X channels. ATP did not affect I(GABA) in neurones: (i) in which P2X channels were not present; (ii) after inhibiting P2X channels with Ca2+ (iii) in the presence of pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid, a P2X receptor antagonist; (iv) after P2X receptor desensitization or (v) at I(ATP) reversal potential. Similarly, GABA did not affect P2X-mediated currents in neurones: (i) in which GABA(A) channels were not present; (ii) in the presence of picrotoxin, a GABA(A) channel blocker; (iii) after GABA(A) receptor desensitization or (iv) at the I(GABA) reversal potential. Current occlusion occurred as fast as current activation and it was still present in the absence of Ca2+, at 11 degrees C, after adding to the pipette solution a cocktail of protein kinase inhibitors (staurosporine + genistein + K-252a), after substituting the GTP in the pipette with GDP-beta-S and after treating the cells with N-ethylmaleimide. Taken together, all of these results are consistent with a model of cross-inhibition between GABA(A) and P2X.     

Origins of cardiac vagal preganglionic fibers: A retrograde transport study

The origin of cardiac preganglionic neurons in the rat was investigated using the retrograde transport of horseradish peroxidase (HRP). A single injection of HRP was made into the right myocardium in either a sinoatrial or mid-ventricular location. Labeled cells were found in the mid- and lower medulla primarily in and around the nucleus ambiguus (NA) 600–1800 μm above the obex. The dorsal motor nucleus of X (DMN) was sparsely labeled and a few cells were found in an intermediate zone near the level of the obex. Labeling was bilateral with slightly heavier labeling found ipsilateral to the injection site than contralateral to it. Following a unilateral vagotomy, labeled cells were only found ipsilateral to the intact vagus. Atrial and midventricular injections yielded similar results. Occasionally only 1– cells in the NA were labeled per section. Inspection of serial sections revealed that in these sparsely labeled rats, the HRP was often in the same location within the NA forming a column of cells within the nucleus. The columns sometimes extended at least 240 μm in the rostral-caudal direction. The columnar organization was most apparent in rats with few labeled cells presumably because it was obscured in nuclei that were heavily labeled.In a second group of rats, the right vagus was cut at the cervical level and dipped in HRP to determine the extent of the NA and DMN in rats. In these animals, heavier labeling was found in the DMN than in the NA. Cells in the DMN were filled from the upper spinal cord to its most rostral extent 1200 μm above the obex. Thus, although the DMN and NA send projections in the vagus nerve, those axons terminating in the myocardium primarily originate in the NA.     

Effect of vagotomy on cholinergic parameters in nuclei of rat medulla oblongata

Cholinergic enzymes and muscarinic receptors in nuclei of rat medulla oblongata were examined after unilateral vagotomy to determine their association with efferent vagal neurons. Vagotomy caused an ipsilateral depletion of acetylcholinesterase from the dorsal motor nucleus of the vagus (DNV) and the nucleus ambiguus (NA). Choline acetyltransferase activity was reduced in ipsilateral DNV, nucleus tractus solitarius and rostral NA. Muscarinic receptor localization by autoradiography with [3H]quinuclidinyl benzilate (QNB) revealed marked intranuclear variations in receptor density. Vagotomy had no effect on the QNB binding pattern. Loss of cholinergic enzymes is a consistent response of motor and preganglionic autonomic neurons to axotomy. Depletion of muscarinic receptors is an additional component of axon reaction in brain stem motoneurons. Accordingly, previous studies have shown a decrease in neurotransmitter-related proteins after axotomy of motoneurons. In the present study, cholinergic enzymes were depleted from axotomized vagal neurons but receptors were not. It is concluded that muscarinic receptors in the DNV and NA are not associated with vagal efferent neurons.     

Expression of c-fos and NGFI-A messenger RNA in the medulla oblongata of the anaesthetized rat following stimulation of vagal and cardiovascular afferents.

Messenger RNA encoding the immediate early genes (IEGs) c-fos and NGFI-A was localized by in situ hybridization of specific 35S-labelled oligonucleotides to detect activated neurones in the medulla oblongata following unilateral electrical stimulation of the vagus (nX) and aortic depressor nerve (ADN), and following mechanical stimulation of the left carotid sinus (CS). In electrically stimulated rats, c-fos and NGFI-A mRNA was strongly expressed in the nucleus tractus solitarius (NTS) (predominantly ipsilaterally), area postrema (AP) and in a dorsal subregion of the paratrigeminal nucleus (PTN). Lower levels of c-fos and NGFI-A mRNA were seen in the ipsilateral NTS and PTN following mechanical stimulation of the left CS. In general these data correlate with the topography of innervation by the different nerve afferents, although the expression in the PTN (and in some cases the AP) would not be predicted on the basis of neuronal innervation patterns reported for the rat. Expression of these IEGs also occurred in the rostral and caudal ventrolateral medulla and inferior olive of both stimulated and sham-operated rats; presumably due to effects of the anaesthesia and surgical procedures. In conclusion the localization of the expression of c-fos and NGFI-A mRNAs represents a useful neuroanatomical technique for detecting the cell bodies of neurones that are activated by cardiovascular nerve afferents and should allow the further characterization of the neurochemical identity of these neurones.     

Ionotropic glutamate receptor subunit immunoreactivity of vagal preganglionic neurones projecting to the rat heart

The ionotropic glutamate receptor subunits expressed by vagal preganglionic neurones in the rat medulla oblongata were examined by using fluorescence immunolabelling combined with retrograde neuronal tracing. The general population of these neurones in the medulla was identified by intraperitoneal injections of Fluorogold and also with choline acetyltransferase antibodies. Cardiac projecting neurones were specifically identified by applying the fluorescent tracer 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine (DiI) to the heart or by injecting cholera toxin B-subunit into the pericardium. Both tracers labelled populations of neurones lying in the dorsal vagal nucleus, intermediate reticular formation and nucleus ambiguus, and when both tracers were applied simultaneously, approximately 50% of cells were dual-labelled. Control experiments established that the labelling was specific for neurones projecting to the heart. Most vagal preganglionic neurones, including those projecting to the heart, irrespective of their location in the medulla, had a similar profile of glutamate receptor immunoreactivity. Labelling of somata for the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic (AMPA) subunit GluR1 was weak or absent, while labelling with antibodies directed to GluR2, a common sequence of GluR2 and GluR3, and GluR4 was moderate or intense. All neurones studied appeared to express the N-methyl-D-aspartate (NMDA) receptor subunit NR1, and while antibodies recognising the NR2A and NR2B splice variants gave strong labelling, immunoreactivity with a NR2B specific antibody was weaker. Weak to moderate labelling was seen in some neurones using antibodies to the kainate receptor subunits KA2 and GluR5-7. These results are consistent with neurophysiological data indicating the presence of AMPA, NMDA and kainate responses in cardiac vagal preganglionic neurones, and suggest that these neurones are similar to other vagal parasympathetic preganglionic neurones in expressing mainly AMPA receptor subunits GluR2-4.     

Changes in expression of NMDA receptor subunits in the rat lumbar spinal cord following neonatal nerve injury

The vulnerability of motoneurones to glutamate has been implicated in neurological disorders such as amyotrophic lateral sclerosis but it is not known whether specific receptor subtypes mediate this effect. In order to investigate this further, the expression of N-methyl-D-aspartate (NMDA) receptor subunits was studied during the first three post-natal weeks when motoneurones are differentially vulnerable to injury following neonatal nerve crush compared to the adult. Unilateral nerve crush was carried out at day 2 after birth (P2) which causes a decrease of 66% in motoneurone number by 14 days (P14). To study receptor expression in identified motoneurones, serial section analysis was carried out on retrogradely labelled common peroneal (CP) motoneurones by combined immunocytochemistry and in situ hybridization (ISH). mRNA levels were also quantified in homogenates from lumbar spinal cords in which the side ipsilateral to the crush was separated from the contralateral side. The NR1 subunit of the NMDA receptor was widely distributed in the spinal cord being expressed most strongly in motoneurone somata particularly during the neonatal period (P3-P7). The NR2 subunits were also expressed at higher levels in the somata and dendrites of neonatal motoneurones compared to older animals. NR2B mRNA was expressed at low to moderate levels throughout the studied period whereas NR2A mRNA levels were low until P21. Following unilateral nerve crush, an initial decrease in NR1 mRNA occurred at one day after nerve crush (P3) in labelled CP motoneurones ipsilateral to the crush which was followed by a significant increase in NR1 subunit expression at 5 days post-injury. This increase was bilateral although reaching greater significance ipsilateral to the crush compared with sham-operated animals. A significant increase in NR1 and NR2B mRNA post injury was also detected in spinal cord homogenates. In addition, the changes in levels of NR1 and NR2B mRNA were reflected by comparable bilateral changes at P7 in receptor protein determined by quantitative immunocytochemical analysis of NR1 and NR2 subunit expression in identified CP motoneurones indicating a co-ordinated regulation of receptor subunits in response to injury.     

Activation of cardiac vagal afferents facilitates late vagal inhibition in neurones of the rostral ventrolateral medulla oblongata bilaterally

A reduced activity of cardiac vagal afferent fibres is considered as one of the pathophysiological causes of post-infarction complications [A. Head, Baroreflexes and cardiovascular regulation in hypertension. J. Cardiovasc. Pharmacol. 26 (1995) S7-S16]. The mechanism of how a reduction of cardiac vagal activity leads to enhanced sympathetic drive and systemic hypertension is however not yet clear. Experimental data have shown that the rostral ventrolateral medulla oblongata (RVLM) plays an important role in tonic blood pressure regulation, the control of sympathetic vasoconstriction and cardiac performance. The aim of the study was to determine whether activation of cardiac vagal afferents contributes to eliciting the long-lasting late inhibition that we have previously shown to occur in neurones of the RVLM [A. Zagon, K. Ishizuka, I. Rocha, K.M. Spyer, Late vagal inhibition in neurones of the ventrolateral medulla oblongata in the rat. Neurosci. 92 (1999) 877-888]. The experiments were carried out in terminally anaesthetised and artificially ventilated rats using in vivo intracellular recordings. The data confirmed that late vagal inhibition is elicited by cumulative activation of functionally different vagal afferents, including those that originate from cardiac receptors. It was also demonstrated that activation of cardiac afferents could lead to a significant increase in the duration of this long-lasting late response component. Facilitation of late vagal inhibition was observed in RVLM neurones both ipsi- and contralateral to the stimulated nerve. It is suggested that such facilitation of late vagal inhibition may be a mechanism of how pulse-synchronous activation of cardiac afferents leads to a tonic modulation of the activity of RVLM neurones. An attenuation of late vagal inhibition during reduced activity of cardiac vagal afferents could lead to enhanced excitability in these neurones which in turn can lead to an increase in medullary sympathetic outflows towards the heart and peripheral blood vessels.     

Estrogen blocks the cardiovascular and autonomic changes following vagal stimulation in ovariectomized rats

The current investigation examines the effect of acute and chronic estrogen administration on baroreflex sensitivity and autonomic tone following 2 h of vagal afferent stimulation in ovariectomized female rats. Female Sprague-Dawley rats were ovariectomized and supplemented daily for 7 days with either estrogen (OVX-E2; 0.5 microg/kg; s.c.) or saline (OVX-S; 0.9%; s.c.). On the 8th day the animals were anaesthetized (sodium thiobutabarbitol; 100 mg/kg) and instrumented for recording blood pressure, heart rate and efferent vagal and renal nerve activities. The baroreflex was evoked using intravenous injection of various doses of phenylephrine hydrochloride (0.025, 0.05+/-0.1 mg/kg). Electrical stimulation of vagal afferents for 2 h produces autonomic imbalance characterized by sympathoexcitation and parasympathetic withdrawal. This protocol of vagal stimulation produced a significant increase in renal nerve activity (from 20+/-6 to 140+/-20 spikes/2 s) and decreases in both vagal nerve activity (from 22+/-3 to 10+/-2 spikes/2 s) and baroreflex sensitivity (from 0.55+/-0.05 to 0.3+/-0.05) in OVX-S female rats. However. vagal stimulation had no effect on baroreflex sensitivity or autonomic nerve activities in OVX-E2 rats. Administration of a single, bolus dose of estrogen (1 x 10(-2) mg/kg) to OVX-S rats immediately prior to termination of vagal stimulation blocked the changes in autonomic nerve activities and baroreflex sensitivity previously observed. These results suggest that both chronic and acute estrogen supplementation may provide resistance to the autonomic disturbances associated with visceral afferent activation.     

P2X receptor-mediated excitatory synaptic currents in somatosensory cortex

Fast P2X receptor-mediated excitatory postsynaptic current (EPSC) was found in pyramidal neurones of layer V of somatosensory cortex in slices acutely isolated from the brain of 17- to 22-day-old rats. The EPSCs were elicited by field electrical stimulation in the layer VI at 0.1 Hz in the presence of picrotoxin. When the glutamatergic EPSC was blocked by glutamate receptors inhibitors NBQX and D-AP5, a residual EPSC (rEPSC) was recorded from 85% of neurones tested. This rEPSC was not affected by blockers of nicotinic (hexamethonium) and serotonin (Y25130) receptors; however, it was reversibly inhibited by P2X receptors antagonists (NF023, NF279, and PPADS). An application of ATP (20 microM), beta,gamma-methylene ATP (25 microM), and alpha,beta-methylene ATP (20 microM) to acutely isolated pyramidal neurones of layer V evoked inward currents (30 to 400 pA) in 75% of cells tested. We concluded that several subtypes of P2X purinoreceptors participate in synaptic transmission in neocortex.     

Induction of c-jun expression in vagal motoneurones following axotomy.

Using in situ hybridization, we have measured c-jun mRNA expression in the caudal medulla oblongata of the rat following sectioning of the cervical vagus nerve. Twenty four hours after either unilateral cervical vagotomy or nodose ganglionectomy, motoneurones in the ipsilateral dorsal motor nucleus of the vagus and nucleus ambiguus expressed increased (2-3 fold) levels of c-jun mRNA. After 7 days, c-jun mRNA expression was further elevated to levels 8-10-fold higher than basal. These results demonstrate an unusual, long-lasting activation of a member of the 'leucine zipper' class of immediate early gene, and suggest that immediate early genes such as c-jun may be responsible for the regulation of the expression of structural protein and neuropeptide genes that have been demonstrated to occur following nerve injury.     

P2X receptors mediate ATP-induced primary nociceptive neurone activation

ATP-gated P2X ion-channel receptors are localised throughout the mammalian nervous system and have been identified on neurones which participate in conduction of nociceptive information from the periphery to, and within, the CNS. This article briefly reviews recently published research describing the role that ATP and P2X receptors may play in pain perception, highlighting the importance of the P2X(3) receptor in this process. The P2X(3) receptor subunit is almost exclusively expressed on a subset of small and medium diameter sensory neurones innervating cutaneous and visceral tissue. Activation of P2X receptors present on the peripheral terminals of primary afferents results in neuronal depolarisation and, in conscious animals, leads to the manifestation of acute nociceptive behaviour. Recent animal studies have also shown that P2X(3) receptor expression is increased in sensory ganglia following acute neuronal injury, hinting that similar plasticity in the expression of this receptor subtype could underlie the mechanisms involved in a range of conditions characterised by sensory hypersensitivity in man. It is apparent from the evidence available that functional antagonists at specific P2X receptor subtypes could represent an important class of novel analgesic agents.