Indomethacin-induced lesion modifies contractile activity in rat small intestines.

BACKGROUND: Indomethacin induces intestinal lesions. The change in contractility of segments with lesions was studied. METHODS: Motility was recorded in segments with lesions isolated from the rat small intestine 24 h after a subcutaneous injection of indomethacin (20 mg/kg). RESULTS: Concentration-contraction curves for acetylcholine shifted leftward independently of the degree of severity of lesions, but the curves for carbachol and 5-hydroxytryptamine did not. Contractions produced by intramural nerve stimulation were enhanced in segments with no visible damage but decreased with progression of lesions. Neostigmine augmented them in normal rats but not in indomethacin-treated rats. The peristaltic activity was enhanced in segments with no visible lesions. CONCLUSIONS: The results suggest that treatment of the rat with indomethacin enhances contractility of the small intestine owing to diminution of acetylcholinesterase activity independently of the degree of lesions and reduces it owing to decreased responsiveness of enteric neurons with the progression of lesions.

Effects of gastrectomy on motility, perfusion pressure, and caerulein-induced relaxation of sphincter of Oddi in dogs.

The effects of subtotal-gastrectomy (gastrectomy) on the spontaneous motility and caerulein-induced relaxation of the sphincter of Oddi (SO) were investigated in the dog. The spontaneous motility and the response to caerulein of the SO were recorded using perfusion method. The basal perfusion pressure (5.1 +/- 0.5 cmH2O) and the frequency of phasic contractions (6.1 +/- 0.5 cycles/min, c/min) of the SO increased to 8.2 +/- 0.6 cmH2O (p < 0.05) and 9.3 +/- 0.4c/min (p < 0.05) after gastrectomy, respectively. They were observed one month after operation (7.8 +/- 0.5 cmH2O and 9.1 +/- 0.9 c/min, p < 0.05), but did not change by vagotomy with sympathectomy (vagosympathectomy). In the spontaneous motility of the SO, the motility index increased to 143.7 +/- 18.7% (p < 0.05) at 4 hrs and 135.0 +/- 9.1% (p < 0.05) at one month after gastrectomy, but did not increase after vagosympathectomy. Caerulein had an inhibitory effect on the SO motility in the normal animal 48.0 +/- 4.2%). Gastrectomy reversed to the excitatory effect from the inhibitory effect to caerulein at 4 hrs (127.6 +/- 5.3%, p < 0.05) and at one month (126.6 +/- 5.3%, p < 0.05) after operation, but not reversed by vagosympathectomy and sham gastrectomy. The excitatory response to caerulein after gastrectomy was not effected by vagosympathectomy. It is concluded that gastrectomy induced the SO dysfunction, an increase of the perfusion pressure and the frequency of phasic contractions of the SO, and a change of the response to caerulein of the SO. These alterations suggests that one of the mechanisms of the regulation of the SO motility exist as the reflex from the stomach and/or uppermost duodenum through intrinsic nervous pathways.

Role of 5-HT3 receptors in peristaltic reflex elicited by stroking the mucosa in the canine jejunum.

1. The role played by the 5-HT3 receptor, a serotonin subtype receptor, in peristaltic reflexes was studied in dogs first given ketamine, then anaesthetized with urethane (1.0 g kg-1, I.V.) and alpha-chloralose (100 mg kg-1, I.V.). The jejunal loop was partitioned into two segments with respect to blood supply. Drugs were infused intra-arterially into each segment. 2. Stroking of the mucosa of the aboral and oral segments elicited an ascending contraction and a descending relaxation, respectively. 3. The ascending contraction was concentration-dependently inhibited by treatment of the aboral segment with the 5-HT3 receptor antagonists ICS 205-930 and ondansetron (1.4 pmol min-1 to 14 nmol min-1 for both). The maximal inhibition was 49.5 and 69.3%, respectively. The response was not affected by treatment of the oral segment with these drugs. The descending relaxation was inhibited by 51.4 and 60.8%, respectively, by treatment of the oral segment with ICS 205-930 and ondansetron (1.4 nmol min-1 for both). 4. The ascending contraction was markedly inhibited by treatment of either segment with hexamethonium (140 nmol min-1). The response was abolished by treating both segments with hexamethonium and by treating the oral segment with atropine (14 nmol min-1). 5. These results suggest firstly that, in the canine jejunum, enteric neurons with 5-HT3 receptors play a role as sensory neurons or interneurons in the ascending excitatory and the descending inhibitory pathways of the peristaltic reflex elicited by stroking the mucosa, and secondly, that the ascending limb is composed of cholinergic interneurons and motoneurons.

Histochemical study of the lumbar colonic nerve supply to the internal anal sphincter and its physiological role in dogs.

The sympathetic innervation of the internal and sphincter (IAS) and its physiological role in maintaining sphincter tone were histochemically and mechanically studied in dogs anesthetized with pentobarbital sodium. Numerous catecholamine-fluorescent nerve fibers with varicosities were identified in the IAS of normal dogs. Such fibers were markedly reduced at one week and one month after resection of the hypogastric nerves (HGNs) or the lumbar colonic nerve (LCN), and disappeared after combined HGN and LCN resection. IAS tone decreased to 37.2% of baseline at 1 h after LCN resection and to 69.9% after HGN resection. It returned to the preoperative level at one week and one month after resection. The restored IAS tone was decreased again by acute transection of the previously intact HGNs or LCN. Combined LCN and HGN resection also caused a marked reduction of IAS tone (36.9%) at 1 h after the procedure, and was then restored to the preoperative level with time. The restored tone was not decreased by phentolamine administration. These findings confirmed that both the LCN and the HGN innervate the IAS and play a physiological role in the development of resting tone. The restoration of IAS tone after denervation may be due to intrinsic myogenic properties of the sphincter.

Ascending contraction mediated by 5-hydroxytryptamine3 receptors in canine small intestine.

We investigated the mechanism of ascending contraction induced by activation of 5-hydroxytryptamine3 (5-HT3) receptors in anesthetized dogs. Pressure-measuring balloons were inserted into a loop of extrinsically denervated jejunum. Drugs were administered via the arterial tree to the oral or the anal segment and the ensuing mechanical responses were monitored. Administration of 2-methyl-5-HT (440 pmol-44 nmol) to the anal segment caused contractions in the oral segment in a dose-dependent manner. This response was inhibited by treating the anal segment with ICS 205-930, cocaine, hexamethonium, or tetrodotoxin and by treating the oral segment with atropine or hexamethonium. The response persisted even after abolition of contraction in the anal segment by nifedipine. These results imply that activation of 5-HT3 receptors can induce an ascending contraction through an enteric excitatory pathway formed by a series of cholinergic interneurons and final cholinergic motor neurons, apart from the anal contraction.

Antroduodenal coordinated contractions as studied by chemical ablation of myenteric neurons in the gastroduodenal junctional zone.

Antroduodenal contractions were studied in rat preparations. Augmented duodenal contractions occurred spontaneously in coordination with antral contractions in normal and saline-pretreated preparations. The coordination did not occur when muscle layers and the myenteric plexus were transversely cut at the duodenum just anal to the gastroduodenal junction. In silent preparations, the coordinated contractions were produced by neostigmine or domperidone. When the antroduodenal junctional zone was pretreated with benzalkonium chloride, the augmented duodenal contractions did not occur spontaneously, and even after administration of neostigmine and domperidone although antral contractions occurred spontaneously. In these preparations, there were notably few myenteric neurons in the junctional zone, but the neurons were distributed normally in the areas where motility was recorded. The results suggest that myenteric neurons mediate antroduodenal coordinated contractions and that the coordination is modified by myenteric cholinergic excitatory and dopaminergic inhibitory pathways.

Capsaicin-sensitive afferents activate a sympathetic intestinointestinal inhibitory reflex in dogs.

1. In urethane-anaesthetized dogs, an intra-arterial infusion of capsaicin (0.7-14 nmol min-1) into a separated jejunal segment inhibited a vagally evoked cholinergic contraction of the other non-infused segments. The mechanism of this reflex was investigated. 2. The inhibition by capsaicin was abolished after bilateral splanchnic nerve section or cervical spinal cord transection (C5 or C6), but was unaffected by bilateral vagotomy. Decerebration partially reduced the inhibition. 3. The inhibition by capsaicin was abolished by pre-treatment with phentolamine or yohimbine, but was unaffected by prazosin or propranolol. 4. Sympathetic efferent discharge of the mesenteric nerve increased with capsaicin application, during which time vagally evoked contractions were inhibited. 5. Single-unit discharges of the major splanchnic and mesenteric afferents increased with capsaicin infusion to the loop which was innervated by the units. 6. Together the results implied that capsaicin stimulated canine intestinal primary afferents, resulting in the sympathetic intestinointestinal inhibitory reflex supraspinally. The inhibition of vagally evoked contractions may be due to a presynaptic inhibition via alpha 2-adrenoceptors, which are activated by the reflex.

Intrinsic reflexes mediated via peptidergic neurons in the smooth muscle esophagus of the hen.

In the extrinsically denervated smooth muscle esophagus of the hen anesthetized with urethane (1 g/kg, i. m.), it was studied whether peptidergic neurons in the intramural plexus are involved in the intrinsic reflex. Ascending and descending contractions, and descending relaxation were induced by electrical stimulation of a narrow segment of the esophagus. Naloxone (1 microM), desensitization to substance P (0.3 microM) and spantide (20 microM) inhibited the ascending and descending contractions, respectively. The degree of the inhibition of the contractile response by a combination of naloxone and substance P was nearly the same as that by a single administration of naloxone or substance P. The ascending and descending contractions were reduced to one-third of the control by hexamethonium (100 microM) and abolished by atropine (10 microM). The descending relaxation was abolished after desensitization to vasoactive intestinal peptide (0.3 microM). Taken together the results suggest that in the hen's esophagus, opioid- and substance P-containing neurons in the intramural plexus may act as preganglionic neurons of cholinergic motor neurons in the ascending and descending excitatory pathways and that vasoactive intestinal peptide-containing neurons are involved in the descending inhibitory pathway.

Antidromic activation of vagal and sympathetic afferents does not produce intestinal contractions in dogs.

The question has been asked whether vagal and sympathetic afferents activated antidromically play a role as motor nerves on the in vivo small intestine in dogs anesthetized with urethane. The vagus nerve of one side was cut above the nodose ganglion and the efferent fibers allowed to degenerate. Peripheral stimulation (5-50 Hz, 0.5-3 ms, 5-25 V) of an intact cervical vagus, being able to excite both efferent and afferent fibers, caused large contractions in the jejunum and stomach, whereas stimulation of the contralateral cut cervical vagus could not produce any response in the jejunum but small contractions in the stomach. Peripheral stimulation of the cut cervical vagus did not produce bradycardia and hypotension. Single- and multi-unit discharges to distension of the jejunal segments could be recorded from the peripheral cut end of the cut cervical vagus. Immunohistochemically, there were many substance P-containing cells in both nodose ganglia. Antidromic stimulation of the dorsal roots (T7-T10) did not induce any response in the jejunum but contractions in the stomach. The results may confirm that vagal and sympathetic afferents have no antidromic motor function at least in the in vivo canine small intestine.

Regulation of bile duct motility by vagus and sympathetic nerves in the pigeon.

Effects of stimulation of the vagus and sympathetic nerves on bile duct peristalses were studied in pigeons anesthetized with urethane. Vagus stimulation increased the frequency of peristalses. Atropine, hexamethonium and tetrodotoxin abolished this excitatory effect. After atropine, inhibition of peristalses sensitive to tetrodotoxin was produced. Stimulation of sympathetic area in the spinal cord inhibited peristalses. Propranolol converted this effect into an excitatory one, which was abolished by phentolamine. The results suggest that vagal and sympathetic innervations of the bile duct in pigeons are similar to those of the sphincter of Oddi in mammalian species.

Myenteric 5-HT-containing neurones activate the descending cholinergic excitatory pathway to the circular muscle of guinea-pig ileum.

1. Participation of myenteric 5-hydroxytryptamine (5-HT)-containing neurones in the ascending and descending pathways of the guinea-pig isolated ileum was investigated in a new preparation. Transmural electrical stimulation of the longitudinal muscle-myenteric plexus (LM-MP) portion of the preparation caused ascending and descending contractions of circular or longitudinal muscle in the attached, intact segments situated orally or anally to the point of stimulation. 2. All contractions of LM-MP stimulation were abolished by tetrodotoxin (0.2 microM). The ascending and descending contractions of circular muscles were also abolished by atropine and inhibited to about 50% by hexamethonium. They were not affected by desensitization to substance P (SP) or by the SP antagonist, (D-Pro2,D-Trp7,9)-substance P. The contractions of longitudinal muscles were inhibited by about 45% by hexamethonium and abolished by a combination of atropine with SP desensitization or the SP antagonist, (D-Pro2,D-Trp7,9)-substance P. 3. Desensitization to 5-HT, ICS 205-930 (1 microM) or cocaine (1 microM) reduced the descending contraction of circular muscle by 80-90%, without significantly affecting the ascending contraction. Methysergide (0.2 microM) failed to alter either contraction. 4. 5-HT desensitization, ICS 205-930 and cocaine only partially reduced the descending contraction of longitudinal muscle. A similar reduction of the ascending contraction (20-30%) was also observed. Methysergide had no effects on either contraction. 5. Contractions of either circular or longitudinal muscle produced by field stimulation of the intact segment were not significantly affected by any of the 5-HT receptor antagonists tested. 6. The results imply that 5-HT-containing neurones, as interneurones, are involved mainly in the descending cholinergic excitatory pathway to the circular muscles.

Involvement of substance P neurons in contractions of canine small intestine produced by mesenteric nerve stimulation.

Pathways for contractions of in vivo canine small intestine produced by mesenteric nerve stimulation (MNS) were studied. In intact and chronically sympathectomized dogs, contractions of jejunal and ileal segments were largely reduced by intra-arterial infusion of capsaicin (10-100 microM, 0.07 ml/min), substance P (SP) antagonist, (D-Pro4, D-Trp7.9) SP (4-11) (100 microM, 0.14 ml/min), hexamethonium (100-1000 microM, 0.07 ml/min) or atropine (100 microM, 0.07 ml/min). In chronically vagotomized dogs, capsaicin, SP-antagonist or atropine significantly reduced MNS-induced contractions, but hexamethonium did not. In dogs in which the coeliac and superior mesenteric ganglia had previously been removed, MNS caused no response although intra-arterial injection of 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP, 0.1 mumol) caused marked contractions. It may therefore be suggested that extrinsic SP neurons probably originating in spinal ganglia and intrinsic SP neurons receiving input from vagal preganglionic cholinergic neurons are involved in the excitatory pathways to MNS-induced contractions and that activation of these neurons excites myenteric cholinergic neurons, thereby causing contractions of the small intestine.

Contractions of the guinea-pig ileum evoked by stimulation of the submucous plexus.

Neural pathways from the submucous plexus to the longitudinal muscle of an adjacent segment of isolated guinea-pig ileum were studied. It was found that electrical field stimulation of a strip of submucosa-submucous plexus produced frequency-dependent longitudinal contractions of an intact segment of intestine lying oral to the point of stimulation. The responses were reduced to less than 10% of control by tetrodotoxin, atropine, morphine and chymotrypsin and by desensitization to substance P (SP). The responses were only inhibited by one-third by hexamethonium and were not affected by desensitization to 5-hydroxytryptamine. The effect of desensitization to SP was reversible, but the effect of chymotrypsin was irreversible. SP-induced desensitization and chymotrypsin did not inhibit the twitch response produced by field stimulation of the whole ileal segment. The same results were observed with preparations made from ileal segments that had been extrinsically denervated. The results suggest that intrinsic neurons with processes in the submucous plexus can excite cholinergic and SP-containing neurons in the myenteric plexus, thereby causing the longitudinal muscle to contract.

Polar innervation of enteric non-cholinergic and non-adrenergic neurons in the isolated guinea-pig ileum.

To study the polarity of the efferent pathway of the myenteric plexus, recordings were made of the mechanical activity of the longitudinal muscle of isolated guinea-pig ileal segments upon stimulation with an electrical field around the myenteric plexus contained within strips of longitudinal muscle (LM-MP) continuous with each end of ileal segment. The amplitude of the contractile response to stimulation of the anal LM-MP was always larger than that to the oral LM-MP. After cholinergic and adrenergic transmission was suppressed by atropine (10 microM) and guanethidine (1 microM), and the tone of the segment was enhanced by histamine (1 microM), the LM-MP stimulation produced non-cholinergic, non-adrenergic (NCNA) ascending contraction and NCNA descending relaxation. The NCNA contraction, but not the NCNA relaxation, was abolished or reduced by desensitization to substance P. The present results suggest that the NCNA innervation of the myenteric plexus participates in the polar effects observed in the guinea-pig ileum, that the NCNA excitatory response may be mediated at least in part by myenteric substance P neurons, and that the NCNA inhibitory response is mediated by non-adrenergic neurons.

Functional role of lumbar sympathetic nerves and supraspinal mechanism in the defecation reflex of the cat.

The role of the lumbar sympathetic nerves and supraspinal mechanism in the defecation reflex was investigated in 30 adult cats and 6 kittens. One or two propulsive contractions, whose mean pressure evoked was more than about 90 cmH2O (adult cats) and 50 cmH2O (kittens), were induced in the rectum of all animals by rectal distension. These propulsive contractions could be generated at the descending and the transverse colons. The removal of the supraspinal influence by spinal transection at T13 or removal of pelvic afferents to the supraspinal center by spinal transection at L abolished the propulsive contractions. Successive lumbar sympathectomy restored the contractions. Lumbar sympathectomy and the successive removal of the supraspinal influence did not affect the propulsive contractions. In both cases, the final exclusion of the sacral segments by pithing of the spinal cord abolished the propulsive contractions. These results suggest that the sacral excitatory reflex mediated via pelvic nerves and the lumbar inhibitory reflex mediated via lumbar sympathetic nerves can function during rectal distension in spinal cats and that the lumbar inhibitory reflex is suppressed by the supraspinal sympathetic inhibitory reflex activated by pelvic afferents in intact cats, as in guinea pigs, resulting in propulsive contractions.

Enteric opioid neurons modulate the basal tone of the isolated puppy ileum.

We studied the role of enteric opioid neurons in the spontaneous motility of the longitudinal muscle in the isolated puppy ileum. Regular fluctuations in tone that rose above and returned to the basal level occurred at an interval of 4.7 +/- 0.3 min. Naloxone (10(-8) and 10(-7) M) reduced the spontaneous tonic contraction by 42.6 +/- 11.6% (p less than 0.02) and 77.0 +/- 3.6% (p less than 0.001), respectively. Tetrodotoxin (3.1 X 10(-7) M) and atropine (10(-7) M) terminated the fluctuations. Met- and Leu-enkephalins (10(-9)-10(-8) M) caused tonic contraction which was abolished by tetrodotoxin and atropine. The contractile response produced by transmural electrical stimulation was reduced by naloxone (10(-7) M). This response was also abolished by atropine and tetrodotoxin. These results suggest that enteric opioid neurons are spontaneously active and might operate, at least in part, to raise the basal tone of the longitudinal muscle in the puppy ileum through a cholinergic excitatory mechanism.