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Simultaneous imaging of Ca2+ signals in interstitial cells of Cajal and longitudinal smooth muscle cells during rhythmic activity in mouse ileum.

Electrical rhythmicity in smooth muscle cells is essential for the movement of the gastrointestinal tract. Interstitial cells of Cajal (ICC) lie adjacent to smooth muscle layers and are implicated as the pacemaker cells. However, the pace making mechanism remains unclear. To study the intercellular interaction during electrical rhythm generation, we visualized changes in intracellular Ca2+ concentration ([Ca2+]i) in smooth muscle cells and myenteric ICC within segments of mouse ileum loaded with a fluorescent Ca2+ indicator, fluo-3. We observed rhythmic [Ca2+]i changes in longitudinal smooth muscle cells travelling rapidly through the smooth muscle cell layer. Between the rhythmic Ca2+ transients, we found brief Ca2+ transients localized to small areas within smooth muscle cells. The amplitude but not the periodicity of rhythmic [Ca2+]i transients in both cell types was partially inhibited by nicardipine, an L-type Ca2+ channel antagonist, suggesting that the rhythmic [Ca2+]i transients reflect membrane potential depolarizations corresponding to both slow waves and triggered Ca2+ spikes. Longitudinal smooth muscle cells and myenteric ICC showed synchronous spontaneous [Ca2+]i transients in eight out of 21 ileac preparations analysed. In the remaining preparations, the synchrony between ICC and smooth muscle cells was absent, although the rhythmicity of the smooth muscle cells was not disturbed. These results suggest that myenteric ICC may play multiple roles including pace making for physiological bowel movement.

Animals↗

An electrical description of the generation of slow waves in the antrum of the guinea-pig.

This paper provides an electrical description of the generation of slow waves in the guinea-pig gastric antrum. A short segment of a circular smooth muscle bundle with an attached network of myenteric interstitial cells of Cajal (ICC-MY) and longitudinal muscle sheet was modelled as three electrical compartments with resistive connexions between the ICC-MY compartment and each of the smooth muscle compartments. The circular smooth muscle layer contains a proportion of intramuscular interstitial cells of Cajal (ICC-IM), responsible for the regenerative component of the slow wave. Hence the equivalent cell representing the circular muscle layer incorporated a mechanism, modelled as a two stage reaction, which produces an intracellular messenger. The first stage of the reaction is proposed to be activated in a voltage-dependent manner as described by Hodgkin and Huxley. A similar mechanism was incorporated into the equivalent cell describing the ICC-MY network. Spontaneous discrete transient depolarizations, termed unitary potentials, are detected in records taken from either bundles of circular smooth muscle containing ICC-IM or from ICC-MY. In the simulation the mean rate of discharge of unitary potentials was allowed to vary with the concentration of messenger according to a conventional dose-effect relationship. Such a mechanism, which describes regenerative potentials generated by the circular muscle layer, also simulated the plateau component of the pacemaker potential in the ICC-MY network. A voltage-sensitive membrane conductance was included in the ICC-MY compartment; this was used to describe the primary component of the pacemaker potential. The model generates a range of membrane potential changes with properties similar to those generated by the three cell types present in the intact tissue.

Animals↗

Factors modifying the frequency of spontaneous activity in gastric muscle.

The cellular mechanisms that determine the frequency of spontaneous activity were investigated in gastric smooth muscles isolated from the guinea-pig. Intact antral muscle generated slow waves periodically; the interval between slow waves was decreased exponentially by depolarization of the membrane to reach a steady interval value of about 7 s. Isolated circular muscle bundles produced slow potentials spontaneously or were evoked by depolarizing current stimuli. Evoked slow potentials appeared in an all-or-none fashion, with a refractory period of approximately 2-3 s. Low concentrations of chemicals that modify intracellular signalling revealed that the refractory period was causally related to the activity of protein kinase C (PKC). Activation of PKC increased and inhibition of PKC activity decreased the frequency of slow potentials. Chemicals that inhibit mitochondrial functions reduced the frequency of slow waves. Inhibition of internal Ca(2+)-store activity decreased the amplitude, but not the frequency of slow potentials, suggesting that the amplitude is causally related to Ca(2+) release from the internal store. The results suggest that changes in [Ca(2+)](i) caused by the activity of mitochondria may play a key role in determining the frequency of spontaneous activity in gastric pacemaker cells.

Animals↗

Autonomic dysfunction in gastrointestinal motility disorders.

The records of 113 consecutive patients with a suspected gastrointestinal motility disorder referred between January 1988 and July 1991 were retrospectively reviewed. The aims were to identify the prevalence of autonomic dysfunction in those with or without associated neurological disease and to determine the diagnostic value of testing for autonomic dysfunction. All patients had gastrointestinal manometry (3 hours fasting, 2 hours fed), 94 of 113 underwent testing of sympathetic adrenergic and cholinergic function and cardiovagal cholinergic function. All tests were scored in a standard manner. There was a significant (p < 0.05) but modest (r = 0.28) rank correlation between autonomic and motility scores. This correlation was stronger (r = 0.67, p = 0.01) in diabetic patients. The number of patients in each group with autonomic dysfunction was as follows: irritable bowel syndrome nine of 33, idiopathic upper gastrointestinal dysmotility six of 21, diabetes mellitus nine of 13, identified non-diabetic neurological syndromes six of nine, postvagotomy or abdominal surgery three of 11, and myopathic pseudo-obstruction two of seven. Autonomic testing is useful in the assessment of autonomic involvement outside the gastrointestinal tract. Logistic discriminant analysis showed that autonomic function testing did not add to the diagnostic value of motility tests in distinguishing between patients with and without irritable bowel syndrome, although a slight improvement was indicated for identifying neuropathic dysmotilities. Thus, the aetiological role of general autonomic dysfunction in irritable bowel syndrome and idiopathic and postvagotomy dysmotilities deserves further study. The addition of autonomic function tests does not add substantially to the diagnostic accuracy of clinical, radiological, endoscopic, and manometric techniques in most patients referred for evaluation of a suspected motility disorder.

Autonomic Nervous System Diseases↗

The effect of flufenamic acid on gastrointestinal myoelectrical activity and transit time in dogs.

BACKGROUND: Flufenamic acid, a fenamate, has been shown to alter markedly the membrane potential of small intestinal smooth muscle and increase intracellular calcium in single cells. AIMS: To determine the effects of flufenamic acid on myoelectrical motor activity and gastrointestinal transit in the intact animal. METHODS: Myoelectrical motor activity was recorded via seromuscular platinum electrodes sutured at regular intervals in the stomach and throughout the small intestine. Fasted and fed gastrointestinal transit was assessed using technetium-99m (99mTc) as the radioactive marker linked to 1 mm amberlite pellets or added to the meal. RESULTS: Flufenamic acid (600 mg, intravenously) induced intense spike activity in the small intestine. The mean duration of irregular spike activity was 250 (7) minutes. Spike activity was more pronounced in the lower small intestine. Flufenamic acid also accelerated initial gastric emptying and markedly shortened transit time in the small intestine. In the fasted state the 50% transit time in the small intestine was 54 (8) minutes with infusion of flufenamic acid compared with 105 (10) minutes in the control group; in the fed state 99mTc first reached the colon at 220 (10) minutes compared with 270 (12) minutes in the control group. CONCLUSIONS: Flufenamic acid had marked effects on both myoelectrical motor complex activity and small intestinal transit in the dog. The observed effects suggest that flufenamic acid may be of potential use as a prokinetic agent.

Animals↗

Involvement of two different pathways in the motor effects of erythromycin on the gastric antrum in humans.

BACKGROUND: During the interdigestive state in humans, erythromycin 40 mg induces a premature activity front that starts in the stomach, while erythromycin 200 mg induces a prolonged period of enhanced antral contractile activity. AIMS: To study the involvement of a cholinergic pathway in the motor effects of erythromycin using the muscarinic antagonist atropine and the neural 5-HT1 receptor agonist sumatriptan. METHODS: In 30 healthy volunteers, fasted antroduodenojejunal motor activity was studied by stationary manometry. Placebo (n = 10), atropine (15 micrograms/kg intravenous bolus plus 15 micrograms/kg/h over 30 minutes; n = 10), or sumatriptan (6 mg subcutaneously; n = 10) was administered, followed by infusion of erythromycin 40 mg or 200 mg. RESULTS: After placebo, erythromycin 40 mg induced a premature activity front with gastric onset after 19.1 (1.7) minutes in all volunteers. After atropine, erythromycin 40 mg failed to induce a premature activity front during a 60 minute period in all volunteers (p < 0.001), while sumatriptan prevented the induction of a premature activity front during a 60 minute period in all but one volunteer (p < 0.005). The number of antral contractions and their mean amplitude in the 60 minutes after erythromycin 200 mg did not differ significantly after atropine or sumatriptan versus placebo. CONCLUSIONS: The antral motor effects of erythromycin in humans are mediated via different pathways. The induction of a premature activity front is mediated through activation of an intrinsic cholinergic pathway, while the induction of enhanced antral contractile activity may be mediated via a pathway potentially involving activation of a muscular receptor.

Anti-Bacterial Agents↗

Impaired development of interstitial cells and intestinal electrical rhythmicity in steel mutants.

Electrical rhythmicity in the gastrointestinal tract may originate in interstitial cells of Cajal (IC). Development of IC in the small intestine is linked to signaling via the tyrosine kinase receptor, c-kit. IC express c-kit protein, and disruption of c-kit signaling causes breakdown in IC networks and loss of slow waves. We tested whether mutations in steel factor, the ligand for c-kit, affect the development of IC networks. IC were found in the region of the myenteric plexus (IC-MY) in mice with steel mutations (i.e., Sl/Sld) at 5-10 days postpartum, but these cells formed an abnormal network. IC-MY were not observed in adult Sl/Sld animals. IC in the deep muscular plexus (IC-DMP) appeared normal in Sl/Sld animals. Electrical slow waves, normally present in the small intestine, were absent in Sl/Sld animals (10-30 days postpartum). Neural inputs were intact in Sl/Sld animals. Steel factor appears important for the development of certain classes of IC, and IC-MY appear to be involved in the generation of electrical rhythmicity in the small intestine.

Animals↗

Spatial and temporal coupling between slow waves and pendular contractions.

In contrast to the mechanisms of segmental and peristaltic contractions in the small intestine, not much is known about the mechanism of pendular contractions. High-resolution electrical and mechanical recordings were performed from isolated segments of the rabbit ileum during pendular contractions. The electrical activities were recorded with 32 extracellular electrodes while motility was assessed simultaneously by video tracking the displacements of 20-40 serosal markers. The electrical activities consisted of slow waves, followed by spikes, that propagated in either the aboral or oral direction. The mechanical activity always followed the initial electrical activity, describing a contraction phase in one direction followed by a relaxation phase in the opposite direction. Pendular displacements were always in rhythm with the slow wave, whereas the direction of the displacements was dictated by the origin of the slow wave. If the slow wave propagated aborally, then the pendular displacement occurred in the oral direction, whereas if the slow wave propagated in the oral direction, then the displacement occurred in the aboral direction. In the case of more complex propagation patterns, such as in the area of pacemaking or collision, direction of displacements remained always opposite to the direction of the slow wave. In summary, the direction and pattern of propagation of the slow wave determine the rhythm and the direction of the pendular motility. The well-known variability in pendular movements is caused by the variability in the propagation of the underlying slow wave.

Animals↗

Mechanisms underlying nutrient-induced segmentation in isolated guinea pig small intestine.

Mechanisms underlying nutrient-induced segmentation within the gut are not well understood. We have shown that decanoic acid and some amino acids induce neurally dependent segmentation in guinea pig small intestine in vitro. This study examined the neural mechanisms underlying segmentation in the circular muscle and whether the timing of segmentation contractions also depends on slow waves. Decanoic acid (1 mM) was infused into the lumen of guinea pig duodenum and jejunum. Video imaging was used to monitor intestinal diameter as a function of both longitudinal position and time. Circular muscle electrical activity was recorded by using suction electrodes. Recordings from sites of segmenting contractions showed they are always associated with excitatory junction potentials leading to action potentials. Recordings from sites oral and anal to segmenting contractions revealed inhibitory junction potentials that were time locked to those contractions. Slow waves were never observed underlying segmenting contractions. In paralyzed preparations, intracellular recording revealed that slow-wave frequency was highly consistent at 19.5 (SD 1.4) cycles per minute (c/min) in duodenum and 16.6 (SD 1.1) c/min in jejunum. By contrast, the frequencies of segmenting contractions varied widely (duodenum: 3.6-28.8 c/min, median 10.8 c/min; jejunum: 3.0-27.0 c/min, median 7.8 c/min) and sometimes exceeded slow-wave frequencies for that region. Thus nutrient-induced segmentation contractions in guinea pig small intestine do not depend on slow-wave activity. Rather they result from a neural circuit producing rhythmic localized activity in excitatory motor neurons, while simultaneously activating surrounding inhibitory motor neurons.

Action Potentials↗

Reflex changes in intestinal tone: relationship to perception.

Using an original technique, we demonstrated a modulation of intestinal tonic muscular activity (intestinal tone) by intestino-intestinal reflexes. In 11 healthy volunteers we quantitated intestinal tone variations as changes in the air volume within a flaccid bag (12 cm long) located in the proximal jejunum and maintained at a constant pressure by an electronic barostat. Validation studies with glucagon showed significant intestinal relaxation (117 +/- 10% delta vol; P less than 0.05). In six healthy volunteers, graded balloon distensions (1 min duration at 10-min intervals in 8-ml stepwise increments) were randomly performed 8 cm orad, 8 cm caudad, and 20 cm caudad to the bag of the barostat. Perception was scored (0-6) by a questionnaire. Distensions at the three sites induced similar perception; at the threshold for discomfort (score greater than or equal to 5) distension also induced intestinal relaxatory responses (43 +/- 10%, 34 +/- 5%, and 32 +/- 4% delta vol from orad to caudad, respectively; P less than 0.05 for all). However, while unperceived orad distensions (13 +/- 2 ml) induced reflex relaxation (21 +/- 6% delta vol; P less than 0.05), 20-cm-caudad distensions at higher levels (16 +/- 2 ml, 2.7 +/- 0.5 perception score; P less than 0.05) did not (1 +/- 7% delta vol). This dissociation between perception and intestinal tone reflexes suggests that both responses to intestinal distension are mediated by specific mechanisms.

Adult↗

Studies in vivo and in vitro on effects of PGE2 on colonic motility in rabbits.

Prostaglandins (PG) of the E series are synthesized throughout the gastrointestinal tract, and their elevated levels have been reported in many diarrheal states, including inflammatory bowel disease. It is already known that PGE2 has region-specific and muscle layer-specific effects in different areas of the intestine. The aim of this study was to evaluate possible dose-related motor effects of PGE2 on rabbit proximal and distal colon both in vivo and in vitro. We found that, in the proximal colon in vivo, PGE2 caused inhibition of myoelectric and mechanical activity at low doses but at higher doses caused marked excitation. Under the same experimental conditions, PGE2 caused only excitation in the distal colon, a phenomenon associated with an increase in antegrade contractions and diarrhea. In vitro, PGE2 caused excitation of both proximal and distal colonic longitudinal muscle and relaxation of the circular muscle. Its actions, however, were much more pronounced in the distal region. It is concluded that PGE2 has profound effects on colonic motility that are concentration dependent and that differ with the region of the colon under study. Furthermore, the evidence also suggests that elevated PGE2 levels in disease states may play a significant role in abnormal colonic motility and may facilitate the onset of diarrhea.

Acetylcholine↗

Effect of erythromycin on gastric myoelectrical activity in normal human subjects.

While a great deal of attention has been paid to the effect of erythromycin (Ery) on gastric contractility, its effect on gastric myoelectrical activity, which controls gastric motility, remains unknown. In this study, Ery (6 mg/kg) was infused intravenously in 14 normal human subjects (placebo controlled). Gastric myoelectrical activity was recorded using the surface electrogastrographic method. The electrogastrogram (EGG) recordings were analyzed using spectral analysis methods. It was found that the presence of the 2-4 cycles/min activity (normal slow wave frequency range) in the EGG was 51 +/- 19% in the first hour of the intravenous Ery infusion, which was significantly smaller (P less than 0.001, t test) than that (72 +/- 20%) during the corresponding control period (intravenous saline). This difference was, however, not quite significant (P = 0.067, t test) in the second hour after the infusions. The average score for nausea during Ery was 4.5 (0 for no nausea, 10 for the most severe nausea). We conclude that 1) intravenous Ery induces irregularities in the cutaneous EGG in normal human subjects; and 2) the noninvasive cutaneous EGG is an attractive method both for the investigation of the effects of pharmacological and prokinetic agents such as Ery on gastric myoelectrical activity in humans and for correlating clinical responses to changes in the EGG.

Adult↗

Symptomatic responses to stimulation of sensory pathways in the jejunum.

We hypothesized that intestinal afferent pathways inducing perception may be selectively activated by transmucosal electrical nerve stimulation, without disruption of the intrinsic myoelectrical rhythm. Hence, in 12 healthy subjects we measured perception (by a questionnaire) and jejunal slow wave activity (by electromyography), and we randomly applied for 1 min at 5-min intervals graded electrical (15 Hz, 100 microseconds) and mechanical stimuli (balloon distension) in the jejunum up to the respective discomfort threshold. Electrical and mechanical stimuli induced dose-related perception; the perception and discomfort thresholds were 39 +/- 7 and 63 +/- 6 mA and 31 +/- 3 and 49 +/- 5 ml for electrical and mechanical stimuli, respectively. More than one-half of electrical stimuli elicited clinical-type symptoms (abdominal pressure, fullness, colicky or sharp sensation) similar to those induced by mechanical stimuli; the remaining electrical stimuli (38 +/- 10%) induced paresthesia or flutterlike sensation. Similar types of symptoms were perceived with weak and strong stimuli. Jejunal slow wave activity (11.3 +/- 0.4 cycles/min) was not modified by either stimuli. We conclude that activation of intestinal sensory pathways, either by transmucosal nerve stimulation or via mechanoreceptors, induces a similar dose-related symptomatic response, without interfering with the intrinsic myoelectrical activity.

Adult↗

Serosal and cutaneous recordings of gastric myoelectrical activity in patients with gastroparesis.

The aims of this study were to 1) investigate gastric myoelectrical activity in patients with gastroparesis, 2) validate the cutaneous electrogastrogram (EGG) in tracking the frequency change of the gastric slow wave, and 3) investigate the effect of electrical stimulation on gastric myoelectrical activity. Gastric myoelectrical activity was recorded in 12 patients with documented gastroparesis using serosal electrodes for > 200 min in each subject. All recordings were made at least 4 days after surgery. Each session consisted of a 30-min recording in the fasting state and a 30-min recording after a test meal. The test meal (liquid or mixed) was selected according to patient's tolerance. Electrical stimulation was performed in three subjects via the serosal electrodes at a frequency of 3 cycles/min. Gastric myoelectrical activity was recorded using serosal electrodes in each session. The serosal recording showed slow waves of 2.5 to 4.0 cycles/min in all 12 subjects. Absence of spikes was noted in 11 of the 12 subjects. The simultaneous serosal and cutaneous recording of gastric myoelectrical activity showed that the frequency of the EGG was exactly the same as that of the serosal recording. Liquid meals resulted in a significant decrease in slow-wave frequency (Student's t test, P = 0.006), and the EGG accurately reflected this change. Electrical stimulation had no effect on the frequency of the gastric slow wave and did not induce spikes.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Intestinal myoelectric response in two different models of acute enteric inflammation.

The inflammatory responses to intraluminal administration of the cytotoxic lectin ricin or the hapten 2,4,6-trinitrobenzenesulfonic acid in 25% ethanol were compared. Myoelectric activity was recorded from in vivo ileal loops in rabbits after administration of either agent. At the end of a 6-h study the loops were removed and prepared for histological evaluation and for ex vivo generation of leukotriene (LT) C4 and prostaglandin (PG) E2. Both agents induced significant increases in mucosal damage, polymorphonuclear leukocyte infiltration, and edema. These changes tended to be patchy in the hapten model and confluent in the ricin model. Spike burst activity and mucosal LTC4 and PGE2 generation were also enhanced significantly. Intraluminal administration of the specific LTD4 antagonist Wy-48252 1 h before either inflammagen, significantly inhibited the myoelectric response without affecting tissue morphology or eicosanoid synthesis. These data demonstrate that there is a stereotypic spike burst response to acute inflammation, which is mediated in part by LTD4.

Acute Disease↗

Esophageal myoelectric responses to magnetic stimulation of the human cortex and the extracranial vagus nerve.

We describe the use of magnetic stimulation of the human motor cortex and the extracranial vagus nerve in the neck to study human esophageal electromyographic (EMG) responses. Diffuse cortical stimulation produced both an early and a late EMG response in the proximal esophagus with latencies of 10.9 +/- 0.5 and 43.4 +/- 4.3 ms, respectively. Increasing the stimulation intensity increased the amplitude and duration, but reduced the latency, of the early response (P < 0.01), whereas the late response remained unchanged. The early response, but not the late response, was facilitated when cortical stimulation was performed during valsalva maneuver. Maximal early response amplitudes were obtained by focal stimulation of either hemisphere 4-8 cm lateral to the vertex. Stimulation of the neck over the extracranial vagus nerve at the angle of the jaw also produced an early and a late EMG response in the proximal esophagus with latencies of 4.3 +/- 0.4 and 56.8 +/- 14.5 ms, respectively. Early responses were also recorded in the midesophagus and distal esophagus from either cortical or vagal stimulation, but amplitudes were smaller and rise times were longer, suggesting passive "volume conduction" from the proximal esophagus. Comparison of laryngeal, pharyngeal, and esophageal early EMG responses showed similar amplitudes and rise times, suggesting that the action potentials were generated locally at each site and that true esophageal responses were being detected.

Adult↗

Equine pelvic flexure myoelectric activity during fed and fasted states.

The pelvic flexure is the midpoint of the equine large colon that marks the junction of dorsal and ventral components. Previous studies of intraluminal pressure in this region indicate that it could be an important motility control center. The present study was undertaken to expand our knowledge of normal myoelectric activity around the pelvic flexure region. Eight bipolar silver wire electrodes were surgically fixed at 5-cm intervals to the colonic serosa of five adult horses, starting 30 cm oral to the pelvic flexure on the left ventral colon and ending 15 cm aboral to the pelvic flexure on the left dorsal colon (LDC). Recordings of myoelectric activity were done after feed had been withheld for 20 h or when the horses had been allowed to eat hay up to the time of the recording session. The activity was recorded on a polygraph, digitized, processed through a commercial graphics software package, and stored on magnetic tape for later analysis. Action potential activity was basically separated into long spike bursts (LSB) that were > or = 4 s duration and short spike bursts (SSB) that were < 4 s duration and quantified by a software program developed exclusively for the equine large intestine.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗