Immunohistochemical localisation of a gastrin-releasing peptide-like material in area postrema, nucleus of the solitary tract and vagal motor nucleus in the brainstem of rat

The presence of an endogenous gastrin-releasing peptide (GRP)-like peptide in the hindbrain of rat was demonstrated immunohistochemically using antisera directed against the N-terminus and C-terminus of GRP. N-terminal and C-terminal-like immunoreactive material were distributed throughout the nucleus of the solitary tract (NTS), dorsal motor nucleus of the vagus (DMV) and tractus solitarius (TS), as well as in areas postrema (AP) and substantia gelatinosa separating AP from NTS. Positive immunostaining was localised to a dense network of nerve fibres which project longitudinally along the neuraxis. Immunolabelled cell bodies were observed rostral to the obex, principally in the mediolateral subnucleus of NTS. These immunopositive neurones project their axons caudally and longitudinally towards the commissural subnucleus of the NTS. Immunolabelled cell bodies also were found in AP; they projected their axons caudally and ventrally towards NTS. Positive immunostaining was blocked by pre-adsorbing antisera with either GRP (1 nmol/ml) or bombesin (3 nmol/ml), but was unaffected by substance P (30 nmol/ml) and spared by capsaicin pretreatments which deplete sensory nerves of their peptide content. The results indicate that NTS neurons containing a GRP-like peptide connect the rostral and caudal regions of the dorsal vagal complex by way of longitudinal nerve tracts descending NTS and TS. Some neurons in AP also contain a GRP-like peptide and appear to connect with the dorsal vagal complex.     

Relationship between the mechanism of gastro-oesophageal reflux and oesophageal acid exposure in patients with reflux disease

Abstract  This study investigated the relationship between the oesophageal acid exposure time and the underlying manometric motor events in patients with gastro-oesophageal reflux disease (GORD). In 31 patients, 3-hour oesophageal motility and pH were measured after a test meal. Ten patients underwent 24-hour ambulatory manometry and pH recording. In the 3-hour postprandial study, of 367 reflux episodes 79% was associated with a transient lower oesophageal sphincter relaxation (TLOSR), 14% with absent basal lower oesophageal sphincter (LOS) pressure and the remaining 7% with other mechanisms, representing 62, 28 and 10% of the acid exposure time, respectively. Acid reflux duration per motor mechanism was longer for absent basal LOS pressure than for TLOSR (189 ± 23 s and 41 ± 5 s, respectively, P  < 0.001). In the 24-hour ambulatory study, the contribution of TLOSRs to reflux frequency vs acid exposure time were 65 vs 54% interprandially and 74 vs 53% after the meal. During the night, absence of basal LOS pressure accounted for 36% of reflux events representing 71% of acid exposure time. In conclusion, the duration of oesophageal acid exposure following a TLOSR is shorter than reflux during absent basal LOS pressure. TLOSRs are, the major contributor to oesophageal acid exposure during the day. At night, however, reflux during absent basal LOS pressure is the major contributor to acid exposure.     

Transient lower oesophageal sphincter relaxations play an insignificant role in gastro-oesophageal reflux to the proximal oesophagus.

Gastro-oesophageal reflux to the proximal oesophagus may cause atypical symptoms of gastro-oesophageal reflux disease (GORD). The motor abnormalities underlying reflux into the proximal oesophagus are still unclear. The aim of this study was to analyse the oesophageal motility during reflux into the proximal oesophagus in a group of healthy subjects and in patients with atypical symptoms of GORD. We concentrated particularly on lower oesophageal sphincter (LOS) activity and transient lower oesophageal sphincter relaxations (TLOSRs). Ten patients (7M, 3F, age 25-51 years) with mild oesophagitis (Savary-Miller grade I-II) and 10 healthy subjects (6M, 4F, age 23-54 years) underwent a 24-h dual pH-metric and manometric recording, using an electronic portable device. This recorded distal and proximal oesophageal pH values, oesophageal body and LOS motility. GORD patients had more distal and proximal reflux (DR and PR) compared with healthy controls (DR P < 0.001; PR P < 0.05). TLOSRs were the most frequent event during reflux into the distal oesophagus, whereas TLOSR frequency was much lower during reflux to the proximal oesophagus in GORD patients and in healthy controls (P < 0.05 and P < 0.01 vs. distal reflux, respectively). A significant relationship between TLOSRs and distal refluxes was present but no relationship with proximal reflux was detected. We conclude that TLOSRs are much less frequent during reflux to the proximal oesophagus than distal oesophageal reflux in patients with mild GORD suffering from atypical manifestations. The mechanism of acid reflux to the proximal oesophagus is unclear.     

Brainstem sites controlling the lower esophageal sphincter and crural diaphragm in the ferret: a neuroanatomical study

The lower esophageal sphincter (LES) and the crural diaphragm (CD) surrounding the esophagogastric junction are key components of the gastroesophageal reflex mechanism, which engages the vago-vagal brainstem circuitry. Although both components work in conjunction to prevent gastroesophageal reflux, little is known about the brain area(s) where this integration takes place. The aims of this study were to: (1) trace the brainstem circuitry associated with the CD and the LES, and (2) determine possible sites of convergence. Experiments were done in adult male ferrets. Under isoflurane anesthesia, recombinant strains of the transneuronal pseudorabies virus (PRV-151 or PRV-Bablu) or the monosynaptic retrograde tracer cholera toxin beta-subunit (CTb) were injected into either the CD or the LES. Following a survival period of 5-7 days, animals were euthanized, perfused and their brains removed for dual-labeling immunofluorescence processing. In animals injected with recombinants of PRV into the CD and the LES, distinct labeling was found in various brainstem nuclei including: area postrema, DMV, nucleus tractus solitarius (NTS), medial reticular formation (MRF) and nucleus ambiguous (NA). Double-labeled cells were only evident in the DMV, NTS and MRF. Injections of CTb into the CD or the LES resulted in retrograde labeling only in the DMV. These findings demonstrate the presence of a direct projection from the DMV to the CD. They further suggest that the neuronal connections responsible for CD or LES function are contained in circuitries that, though largely independent, may converge at the level of DMV, NTS and MRF.     

Mechanisms of gastro-oesophageal reflux in the ferret

Transient lower oesophageal sphincter (LOS) relaxation is the major mechanism of gastro-oesophageal reflux in humans – an event unassociated with swallowing. Mechanisms involved in triggering transient LOS relaxation are poorly understood, and their further study requires a small animal model. In this study we aimed to establish methods for prolonged ambulant oesophageal manometry in ferrets, and to determine motor events associated with reflux episodes and their triggering by different gastric nutrient loads. Forty-two studies were performed on nine ferrets with chronic cervical oesophagostomies, through which a manometric assembly was introduced and secured to a collar, which incorporated a microphone for detection of swallows. The assembly included a gastric feeding channel, one gastric and four oesophageal manometric sideholes, a 2.5-cm-long LOS sleeve sensor, and an oesophageal pH electrode. Intragastric infusions were given over 2 min, the first after a 30-min control recording period, and in 29/42 studies, a second infusion was given 60 min later. Infusions were either 25 mL 10% dextrose solution, pH 3.5 (22 studies), 25 mL triglyceride emulsion (Intralipid) pH 3.5 (11 studies), or 25 mL air (nine studies). Episodes of oesophageal acidification were absent before gastric infusions. After infusion, 2.1 ± 0.2 episodes occurred over the first 30 min. After glucose infusion, 15/18 acidification episodes (83%) occurred during transient LOS relaxation, and 3/18 (17%) occurred after gradual (< 1 mmHg sec⁻¹) downward drifts in basal LOSP to < 2 mmHg. After lipid infusion two acidification episodes occurred, both during transient LOS relaxation. Mean duration of transient LOS relaxation was 8.0 ± 0.4 sec. All infusions increased occurrence of transient LOS relaxation to a similar extent, each of which ended with primary peristalsis. We conclude that gastric infusion of glucose, lipid and gas are all effective in provoking gastro-oesophageal reflux in ferrets. Reflux occurs through similar mechanisms to those seen in humans, i.e. increased triggering of transient LOS relaxation. The conscious ferret is therefore an appropriate model for future studies of manipulation of mechanisms giving rise to gastro-oesophageal reflux.     

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.     

Transient lower oesophageal sphincter relaxation in achalasia: everything but LOS relaxation

Abstract  In conducting clinical high-resolution oesophageal pressure topography (HROPT) studies we observed that after subjects sat upright between series of supine and upright test swallows, they frequently had a transient lower oesophageal sphincter relaxation (TLOSR). When achalasia patients were studied in the same protocol, they exhibited a similar HROPT event leading to the hypothesis that achalasics had incomplete TLOSRs. We reviewed clinical HROPT studies of 94 consecutive non-achalasics and 25 achalasics. Studies were analyzed for a TLOSR-like event during the study and, when observed, that TLOSR-like event was characterized for the degree and duration of distal oesophageal shortening, the degree of LOS relaxation, associated crural diaphragm (CD) inhibition, oesophageal pressurization and upper oesophageal sphincter (UOS) relaxation. About 64/94 (68%) non-achalasics and 15/24 (63%) of achalasics had a pressure topography event after the posture change characterized by a prolonged period of distal oesophageal shortening and/or LOS relaxation. Events among the non-achalasics and achalasics were similar in terms of magnitude and duration of shortening and all were associated with CD inhibition. Similar proportions had associated non-deglutitive UOS relaxations. The only consistent differences were the absence of associated LOS relaxation and the absence of HROPT evidence of reflux among the achalasics leading us to conclude that their events were incomplete TLOSRs. Achalasic patients exhibit a selective defect in the TLOSR response suggesting preservation of all sensory, central and efferent aspects of the requisite neural substrate with the notable exception of LOS relaxation, a function of inhibitory (nitrergic) myenteric plexus neurons.     

The effect of glucagon-induced gastric relaxation on TLOSR frequency

This study aimed to determine the effect of glucagon-induced gastric relaxation on the frequency of transient lower oesophageal sphincter relaxations (TLOSRs). Eight normal subjects (four male, age 18-52 y) were studied after a 6-h fast using a combined manometric barostat assembly. The recording was divided into two 1-h sessions: (1) a baseline period with the barostat set at minimal distending pressure (MDP) + 2 mmHg and (2) a period with continuous glucagon or placebo infusion with barostat set at MDP + 2 mmHg. Patients were studied on two different days and randomly received glucagon (4.8 microg kg(-1) bolus followed by 9.6 microg kg(-1) h(-1) infusion) on 1 day and placebo (saline) on another. Lower oesophageal sphincter (LOS) pressure, frequency of TLOSRs, and barostat bag volumes were determined for both placebo and glucagon infusion. Glucagon induced significant fundal relaxation compared with placebo (P < 0.05) and significantly decreased baseline LEOS pressure (P < 0.05). The frequency of TLOSRs was not altered by glucagon infusion compared with placebo. Despite causing substantial proximal stomach relaxation, glucagon did not increase TLOSR frequency. This suggests that the relevant gastric mechanoreceptors responsible for triggering TLOSRs do not respond to passive elongation.     

Excitatory and inhibitory local circuit input to the rat dorsal motor nucleus of the vagus originating from the nucleus tractus solitarius

The nucleus tractus solitarius (NTS) and dorsal motor nucleus of the vagus nerve (DMV) constitute sensory and motor nuclei of the dorsal vagal complex, respectively. We used whole-cell patch-clamp recordings from DMV neurons in rat brain slices and three methods of stimulation (electrical, glutamate microdrop, glutamate photostimulation) to test the hypothesis that convergent excitatory and inhibitory inputs to DMV neurons originate from intact neurons in multiple NTS areas. Electrical stimulation of the NTS resulted in evoked excitatory and inhibitory postsynaptic currents (eEPSCs and eIPSCs) in DMV neurons. Stimulation of the dorsal NTS with glutamate microdrops, which selectively stimulates the soma and dendrites of intact neurons, resulted in 31% of DMV neurons receiving eEPSCs, 44% receiving eIPSCs, and 6% receiving convergent excitatory and inhibitory inputs. Glutamate photostimulation allowed selective activation of intact neurons in multiple, discrete areas of the NTS and resulted in 36% of DMV neurons receiving eEPSCs, 65% receiving eIPSCs and 20% receiving both inputs. Data obtained by stimulation of multiple NTS areas support the hypothesis that there are anatomically convergent inputs to DMV neurons originating from intact neurons within the NTS. These data support the hypothesis that there is transfer of convergent information from the NTS to the DMV, implying that significant sensory-motor processing occurs within the brainstem.     

Release of nitric oxide in the central nervous system mediates tonic and phasic contraction of the cat lower oesophageal sphincter

Nitric oxide (NO) in the brainstem is implicated in the control of swallowing and oesophageal peristalsis. This study examines the role of brainstem NO in the maintenance of lower oesophageal sphincter (LOS) tone, relaxation and contraction. In urethane-anaesthetized cats, oesophageal peristalsis and sphincter pressures were continuously monitored. Drugs were administered into the fourth ventricle. Oesophageal peristalsis and sphincter relaxation and contraction were induced by superior laryngeal nerve stimulation or intra-oesophageal balloon distention. Basal sphincter pressure was significantly reduced after the i.c.v. administration of the nitric oxide synthase (NOS) inhibitor, l-Ng-monomethyl arginine. The inhibitor's d-isomer had no significant effect on basal sphincter pressure, while l-arginine partially reversed the effect. The NOS inhibitor had no effect on sphincter relaxation, whereas the contraction of the sphincter following relaxation was significantly inhibited. Central nitric oxide synthase inhibition reduces basal LOS tone and contraction amplitude but has no effect on swallow or balloon distention induced sphincter relaxation. Therefore, central release of NO acts in the pathway to stimulate dorsal motor nucleus of the vagus neurones projecting to excitatory neurones in the sphincter. Inhibition of nitric oxide synthase in the CNS does not prevent relaxation of the LOS, suggesting that other pathways that do not utilize NO are important in the induction of LOS relaxation.     

Computer analysis of prolonged lower oesophageal sphincter pressure recordings

The aim of the study was to validate a recently developed computer program for the analysis of prolonged recordings of lower oesophageal sphincter pressure. Thirty 1-hour stretches were selected from sets of 24-h pressure signals recorded from the pharynx, oesophagus, lower oesophageal sphincter (LOS) and stomach in 10 ambulant patients with gastrooesophageal reflux disease. Three experienced investigators visually analysed end-expiratory LOS pressures and transient lower oesophageal sphincter relaxations (TLOSRs), using published criteria. A computer program was developed for calculation of an end-expiratory pressure curve and detection of TLOSRs using the same criteria. Although the results showed an maximum deviation from the mean of 11.1% and 14.8% for manually calculated LOS pressures and visually detected TLOSRs, respectively only 62.1% of the detected TLOSRs were detected by all three observers. LOS pressure as measured by the computer closely approximated the mean of the LOS pressures calculated by the three observers. Although the total number of TLOSRs was comparable to that assessed by visual analysis, the computer detected only 46% of the TLOSRs detected by each observer and 56.8% of the TLOSRs detected by all observers. It is concluded that automated calculation of end-expiratory LOS pressure is feasible and yields reliable results, whereas automated detection of TLOSRs could not be satisfactorily accomplished. Our study showed that improvement of computer algorithms for TLOSR detection is desirable. However, the previously described criteria for detection of TLOSRs are insufficiently precise; further refinement of these criteria will be necessary to reduce the large discrepancies between the outcome of detection of TLOSRs by computer and by humans, and to reduce the equally large discrepancies between the results of detection by different human observers.     

Vagal ganglionic and nonadrenergic noncholinergic neurotransmission to the ferret lower oesophageal sphincter

In the present study we aimed to discretely characterise ganglionic and neuroeffector transmission to the ferret lower oesophageal sphincter (LOS) using a novel preparation of LOS muscle with intact vagal innervation in conjunction with isolated LOS muscle strips. In this way we could compare vagally mediated LOS relaxation with that of enteric inhibitory motorneurones which were directly stimulated. Preparations of LOS muscle, with or without attached vagus nerves, were dissected from adult ferrets and maintained under preload in organ baths, where LOS muscle developed spontaneous tone. LOS relaxations in response to vagal stimulation (0.5-5 Hz, 30 V) were recorded, alone and following pretreatment with tetrodotoxin (TTX), hexamethonium (Hex), Hex and atropine and NG-nitro-L-arginine (L-NNA). Direct activation of enteric inhibitory motorneurones was performed via electrical field stimulation (EFS). Vagal stimulation elicited frequency-dependent relaxations of the LOS that were abolished by tetrodotoxin (1 microM) and markedly reduced following L-NNA pretreatment (100 microM), but unaltered following pretreatment with the selective VIP or PACAP receptor antagonists VIP (10-28) or PACAP (6-38), respectively (each at 5 microM). The potent NOS inhibitor S-methyl-L-thiocitrulline (100 microM) inhibited LOS relaxation to the same degree at 5 Hz. Hex alone (500 microM) reduced maximal relaxation by 50%; in combination with atropine (2 microM), relaxation was almost abolished. In isolated LOS muscle strips, neither VIP (10-28) nor PACAP (6-38) altered EFS-induced relaxation. Taken together, these results suggest ganglionic neurotransmission to the ferret LOS occurs mainly through a combination of nicotinic and muscarinic receptors and utilises nitroxidergic enteric inhibitory motorneurones to relax the LOS. Moreover, LOS relaxation due to direct activation of inhibitory motorneurones also utilises primarily nitric oxide and other as yet undefined neurotransmitters. Neither VIP nor PACAP are involved in vagally mediated or direct enteric neuronally stimulated LOS relaxation in the ferret.     

Mapping the effects of motor cortex stimulation on single neurons in the dorsal column nuclei in the rat: direct responses and afferent modulation

The major aim of this study was to define the topography of descending motor cortical influences on the dorsal column nuclei of the rat. A total of 1442 single neurons were recorded throughout the rat cuneate (Cu) or external cuneate (ECu) nuclei and classified according to their: 1) location within the nuclei, 2) receptive fields, and 3) response to standardized deep layer microstimulation in the forelimb area of the motor (MI) cortex. Excitatory neuronal responses to this MI cortical stimulation ranged in latency from 2-28 msec, with 80% of responses in the 4-12 msec range. Overall, the rostral extremity of the Cu (0.5-1.0 mm rostral to the obex), and the ECu contained the highest percentage of recorded neurons responding to the MI cortical stimulation (61%). By contrast, only 11% responded in the middle subregion of the Cu (+0.4 to -0.9 mm relative to the obex), and 28% responded in the caudal-most subregion (1.0-2.0 mm caudal to obex). A similar paradigm was used to investigate the topography of MI corticofugal modulation of afferent transmission through the Cu and ECu. The MI cortical stimulation was used as a conditioning (C) stimulus in a condition-test (C-T) paradigm in which the test (T) stimulus was standardized mechanical vibration of digit No. 4 of the contralateral forepaw. A total of 30 cells from different subregions were analyzed by measuring the averaged evoked unit responses to the T-stimuli, and plotting them as a function of C-T intervals from 0-50 msec.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of endotoxin on opossum oesophageal motor function

Endotoxin induces nitric oxide (NO*) synthase and alters gastrointestinal functions. We explored the effect of lipopolysaccharide (LPS) on oesophageal motor function at 6, 12, 24, and 48 h. The effects of inhibiting inducible NO* synthase (iNOS) were studied 12 h after administration of LPS with/without aminoguanidine (AG). Oesophageal manometry was performed and tissue bath studies were performed with muscle strips from the oesophagus and lower oesophageal sphincter (LOS). Plasma nitrite/nitrate concentrations were determined. The amplitudes of peristaltic pressure waves, resting LOS pressure and the percentage LOS relaxations were diminished by LPS. AG attenuated the decrease in amplitude of oesophageal pressure waves, LOS pressure, and percentage relaxation of LOS brought about by LPS. LPS decreased electrical field stimulation (EFS)-induced relaxation of LOS muscle. AG attenuated this decrease in LOS relaxation. The off-response of transverse oesophageal muscle strips was decreased, and AG antagonized this effect. Plasma concentrations of nitrite/nitrate were increased. The increase in plasma nitrite/nitrate was attenuated by AG. These studies support the hypothesis that endotoxin modulates oesophageal motor function by increasing NO production and suggest that this results from the induction of iNOS.     

Distinct patterns of oesophageal shortening during primary peristalsis, secondary peristalsis and transient lower oesophageal sphincter relaxation

This study characterized oesophageal shortening during secondary peristalsis and transient lower oesophageal sphincter relaxation (TLOSR) in an attempt to determine its contribution to the opening mechanism. Eight healthy subjects (four males, 26 +/- 1 years) had metal clips affixed at 0, +3, and +8 cm relative to the squamocolumnar junction (SCJ), defining two distal oesophageal segments. Axial clip movement was assessed with concurrent videofluoroscopy and manometry during primary peristalsis, secondary peristalsis and TLOSR. Clip-defined oesophageal segment length change was measured at 0.5-s intervals. The magnitude of the most distal segment shortening was least with TLOSR, greatest with primary peristalsis and intermediate with secondary peristalsis. Conversely, maximal overall oesophageal shortening during TLOSR, evidenced by SCJ movement, was similar to that during primary peristalsis. In 3/12 TLOSRs, the moment of LOS opening and gas reflux was optimally imaged; SCJ excursion was 0.3 +/- 0.1 cm prior to LOS opening and 1.4 +/- 0.7 cm immediately after gas reflux. The segmental pattern of oesophageal shortening was distinct during primary peristalsis, secondary peristalsis and TLOSR. During TLOSR, significant elevation of the SCJ occurred only after LOS opening, suggesting that this was a consequence of oesophageal distension induced by gas reflux rather than a component of the opening mechanism.     

The effect of mechanoreceptor stimulation of the laryngopharynx on the oesophago-gastric junction.

Stimulation of the pharynx by injection of minute amounts of water induces prolonged period of lower oesophageal sphincter (LOS) relaxation and augmentation of the crural diaphragm (CD) contraction. The purpose of this study was to determine the effects of stimulating mechano-receptors at the laryngopharynx on the LOS and CD by using an air pulse stimulation device. Air pulses were delivered via the internal channel of a flexible endoscope. Oesophageal, LOS, and CD pressures; CD electromyogram; and oesophageal pH 5 cm above the LOS were recorded in nine healthy subjects. Stimulation of the laryngopharynx induced LOS relaxation in the absence of swallow and oesophageal peristalsis. The stimulation of epiglottis and arytenoid produced higher incidence of LOS relaxation compared to the base of tongue. The magnitude of LOS relaxation differed significantly between the three anatomical sites, with greater relaxation occurring at the epiglottis and arytenoid compared with the base of the tongue. None of the LOS relaxations induced by laryngeal stimulation resulted in inhibition of the CD or gastroesophageal reflux. We conclude that stimulation of the laryngopharyngeal mechanoreceptors induces LOS but not CD relaxation. The LOS relaxation induced by laryngopharyngeal stimulation is not accompanied by acid reflux in fasting state.     

Bulbospinal projections to the intermediolateral cell column: a neuroanatomical study

The location of those neurones in the brain stem that project to the intermediolateral column (ILC) from which preganglionic sympathetic neurones have their origin was studied by the method of retrograde transport of horseradish peroxidase (HRP). In cats 30--50 nl of a 30% HRP solution was injected into the region of the ILC at T3 or L1 on one side. After a survival period of 72 h the lower brain stem from C1 to the inferior colliculi was sectioned and prepared for histological study under brightfield illumination. Neurones stained with exogenous HRP were found in three regions: (a) in the ipsilateral, dorsomedial part of the nucleus of the solitary tract (NTS) (43% of all labeled neurones), in the cranial part of the NTS, and also on the contralateral side (7%); (b) in the ventrolateral reticular formation beginning at the level of the obex up to 8 mm cranial to the obex (25% ipsilateral, 3% contralateral); and (c) in the ventral part of the raphe nuclei (postpyramidal and inferior central nucleus) from 2 to 9 mm cranial to the obex (22%).     

Similarities of the neuronal circuit for the induction of fictive vomiting between ferrets and dogs

Previous studies suggested that the following neuronal circuit participates in the induction of vomiting by afferent vagal stimulation in decerebrated paralyzed dogs: (1) afferent fibers of the vagus nerve, (2) neurons of the solitary nucleus (NTS), (3) neurons of the prodromal sign center near the semicompact part of the nucleus ambiguus (scAMB), (4) neurons of the central pattern generator in the reticular area adjacent to the compact part of nucleus ambiguus (cAMB), (5) respiratory premotor neurons in the caudal medulla, (6) motor neurons of the diaphragm and abdominal muscles. However, the commonality of this neuronal circuit in different species has not yet been clarified. Thus, this study was conducted to clarify this point. This study clarified for the first time that fictive vomiting in decerebrated paralyzed ferrets could be induced by vagal stimulation, and could be identified by centrifugal activity patterns of the phrenic and abdominal muscle nerves. The distributions of c-Fos immunoreactive neurons in the NTS, scAMB and cAMB areas in ferrets that exhibited fictive vomiting were denser than those in ferrets that did not. Application of the nonNMDA receptor antagonist into the 4th ventricle produced the reversible suppression of fictive vomiting. The NK1 receptor immunoreactive puncta were found in the reticular area adjacent to the scAMB. Microinjections of NK1 receptor antagonist into the reticular areas on both sides abolished fictive vomiting. All these results in the ferrets are identical with results previously obtained in dogs and cats. Therefore, this suggests that the above neuronal circuit commonly participates in the induction of emesis in these animal species.     

The lower oesophageal sphincter

Abstract  The lower oesophageal sphincter (LOS) is a specialized segment of the circular muscle layer of the distal oesophagus, accounting for approximately 90% of the basal pressure at the oesophago-gastric junction. Together with the crural diaphragm, it functions as an antireflux barrier protecting the oesophagus from the caustic gastric content. During swallowing or belching, the LOS muscle must relax briefly in order to allow passage of food or intragastric air. These swallow-induced and prolonged transient lower oesophageal sphincter relaxations (TLOSRs) respectively result from activation of the inhibitory motor innervation of the sphincter. Both in man and animals, the main neurotransmitter released by the inhibitory neurones is nitric oxide. The two typical examples of dysfunction of the LOS are achalasia and gastro-oesophageal reflux disease (GORD). Achalasia is characterized by reduction or even absence of the inhibitory innervation to the LOS, leading to impaired LOS relaxation with dysphagia and stasis of food in the oesophagus. On the contrary, GORD results from failure of the antireflux barrier, with increased exposure of the oesophagus to gastric acid. This leads to symptoms such as heartburn and regurgitation, and in more severe cases to oesophagitis, Barrett's oesophagus and even carcinoma. To date, TLOSRs are recognized as the main underlying mechanism, and may represent an important target for treatment. More insight in the pathogenesis of both diseases will undoubtedly lead to new treatments in the near future.     

Medullary cells of origin of vagal cardioinhibitory fibers in the pigeon. II. Electrical stimulation of the dorsal motor nucleus

Stimulation of the peripheral end of the cervical vagus results in marked bradycardia, a secondary fall in blood pressure, and little change in respiration. Stimulation of the central end results in immediate apnea, a moderate fall in blood pressure, and little change in heart rate. Electrical stimulation of the dorsal medulla produces short-latency bradycardia at points clustering in: (a) lateral dorsal motor nucleus, 0.5–1.0 mm rostral to the obex; (b) solitary tract; and (c) commissural nucleus of Cajal. Apnea frequently accompanies bradycardia except when elicited by stimulation of the dorsal motor nucleus. Hypotension always follows bradycardia, while hypertension accompanies tachycardia. These bradycardia responses are abolished by bilateral vagotomy and severly diminished when the ipsilateral vagus is cut or pharmacologically blocked. Beta-sympathetic blockade has no effect on the occurrence of bradycardia responses. It is concluded that an efferent cardioinhibitory region in the pigeon is localized in the lateral dorsal motor nucleus 0.5–1.0 mm rostral to the obex and that bradycardia elicited from the solitary tract and commissural nucleus result from stimulating afferents. These results correlate nicely with the localization of the cells of origin of vagal cardioinhibitory fibers based on retrograde degeneration experiments, and together the anatomical and physiological findings indicate a concentration of these neurons in the ventrolateral aspect of the dorsal motor nucleus approximately three-quarters of a millimeter rostral to the obex.