Development of interstitial cells of Cajal and pacemaking in mice lacking enteric nerves.

BACKGROUND & AIMS: Development of interstitial cells of Cajal (ICC) requires signaling via Kit receptors. Kit is activated by stem cell factor (SCF), but the source of SCF in the bowel wall is unclear and controversy exists about whether enteric neurons express the SCF required for ICC development. METHODS: Glial cell line-derived neurotrophic factor (GDNF) knockout mice, which lack enteric neurons throughout most of the gut, were used to determine whether neurons are necessary for ICC development. ICC distributions were determined with Kit immunofluorescence, and function of ICC was determined by intracellular electrical recording. RESULTS: ICC were normally distributed throughout the gastrointestinal tracts of GDNF-/- mice. Intracellular recordings from aganglionic gastrointestinal muscles showed normal slow wave activity at birth in the stomach and small intestine. Slow waves developed normally in aganglionic segments of small bowel placed into organ culture at birth. Quantitative polymerase chain reaction showed similar expression of SCF in the muscles of animals with and without enteric neurons. Expression of SCF was demonstrated in isolated intestinal smooth muscle cells. CONCLUSIONS: These data suggest that enteric neurons are not required for the development of functional ICC. The circular smooth muscle layer, which develops before ICC, may be the source of SCF required for ICC development.     

Role of the longitudinal smooth muscle coat in the ileal motile activity: evidence of ileo-ileal inhibitory reflex

BACKGROUND: All gut movements are claimed to be activated essentially by the concentric contraction of the circular muscle, moving the chyme aborally. The role of the longitudinal smooth muscle of the small intestine in gut motility is poorly understood; this point was investigated in the current study. METHODS: The abdomens of 14 crossbreed dogs (eight dogs, six bitches) were opened. A segment of the small intestine was distended by a balloon in increments of 2 mL of saline, and the pressure and electrical activity were recorded proximally and distally to the balloon. The gut wall around the balloon was anesthetized and the test was repeated. The longitudinal muscle coat of the small intestine segment was then excised, and the pressure response and electrical activity were recorded on ileal distension. RESULTS: Two milliliter ileal distension produced pressure decrease (P < 0.05) proximally and distally to the balloon and caused balloon movement. Four, 6 and 8 mL distension effected similar pressure response, while 10 mL showed no response. Electrical waves were recorded from the three electrodes applied to the ileal segment. Upon ileal distension, electrical activity increased over the distended area, with no activity proximally and distally to it. Balloon distension of the anesthetized ileal segment produced no pressure response or electrical activity. After longitudinal myectomy, no electrical activity was recorded at rest or upon ileal distension, and the balloon did not move. CONCLUSION: Ileal distension initiated circular muscle contraction only in the presence of the overlying longitudinal muscle, which appears to transmit the electrical activity to the circular muscle upon ileal distension. Ileal contraction is suggested to initiate ileal hypotonia in the proximal and distal ileal segments mediated through an 'ileo-ileal inhibitory reflex' that leads to aboral progress of the proximally and distally located chyme.     

Gastrointestinal myoelectric activity in an infant with congenital idiopathic motility disorder

We investigated myoelectric activity in an 8-month-old male who presented with a perinatal bowel obstruction, duodenal band, congenital short small intestine, and persistent feeding intolerance. Serosal electrodes were surgically implanted on stomach, duodenum, and jejunum during Nissen fundoplication and ileostomy. A 5-cm ileal specimen was taken for in vitro studies. Spontaneous migrating myoelectric complexes (MMC) were present in stomach and small intestine. Bethanechol increased electrical response activity (ERA) in stomach and duodenum. Morphine induced intense ERA and distinct phase III activity. Pentagastrin infusion did not disrupt MMC cycling. Feeding disrupted MMC complex cycling 30-40 min after the meal. Metoclopramide before feeding delayed disruption of the MMC cycling after the feeding. Intermittent gastric arrhythmias were present after the fifth postoperative day. In vitro muscle strips showed spontaneous contractions and electrical control activity (ECA). Bethanechol, McNeil A-343, motilin, and cholecystokinin induced contractions, but pentagastrin had no effect. We conclude that in spite of a major clinical motility dysfunction, several of our findings were normal. The abnormalities include short MMC period, absence of disruption of MMC by pentagastrin, and gastric arrhythmias.     

Neuromuscular and vascular hamartoma of small bowel presenting as inflammatory bowel disease.

We present the case of a rare hamartomatous condition of the small intestine clinically mimicking inflammatory bowel disease. Disorganised fascicles of smooth muscle derived from both muscularis mucosae and propria, bundles of non-myelinated nerve fibres with groups of abnormal ganglion cells, and haemangiomatous vessels were present within the submucosa of a long segment of small intestine causing subacute obstruction.     

Neuromuscular and vascular hamartoma of small bowel

Two examples of a hitherto undescribed hamartomatous condition of the small intestine are presented. They consist of fascicles of smooth muscle derived from the muscularis mucosae, bundles of unmyelinated nerve fibres with scattered ganglion cells, and haemangiomatous vessels, occurring focally within a segment of the intestine, and causing stenosis.     

Interstitial cells of cajal and inflammation-induced motor dysfunction in the mouse small intestine

BACKGROUND & AIMS: Interstitial cells of Cajal (ICC) play an important role in the control of gastrointestinal motility. We aimed to determine a potential role for ICC in the pathophysiology of inflammation-induced motor disorders. METHODS: Effects of Trichinella spiralis infection on electrical pacemaker activity, the structure of ICC associated with Auerbach's plexus, and in vivo motor patterns were studied in the mouse small intestine. RESULTS: Between days 1 and 15 after infection, structural damage occurred in the network of ICC, particularly in the processes connecting ICC to each other and to smooth muscle cells. This was associated with desynchronization of electrical pacemaker activity. Abnormal slow wave activity occurred, including doubling of frequency and electrical quiescence, leading to the development of ectopic pacemaker activity in vivo. In vivo motor patterns in the small intestine changed from consistent peristaltic contractile activity in control animals to periods of quiescence alternating with both orally and aborally propagating contractile activity in the presence of inflammation. Sixty days after infection, all parameters studied had returned to normal values. CONCLUSIONS: Inflammation-induced alterations in the network of ICC of the small intestine associated with Auerbach's plexus lead to disorganization of motor patterns. Because of the strong temporal correlation between damage to the ICC network, electrical uncoupling, the appearance of ectopic pacemaker activity, and the occurrence of retrograde peristalsis, it is concluded that ICC can play a major role in inflammation-induced motor disturbances.     

Gastric electrical stimulation in patients with gastroparesis

Abstract As in the heart, there is a pacemaker in the human stomach and it generates myoelectrical activity with a frequency of approximately three cycles per minute. Abnormalities in gastric myoelectrical activity may result in gastric motility disorders, such as gastroparesis. Electrical stimulation of the stomach is achieved by delivering electrical currents via electrodes attached to the smooth muscle of the stomach. Recently, a number of studies on electrical stimulation of the stomach in both humans and dogs have indicated that gastric electrical stimulation with appropriate parameters is able to entrain gastric slow waves and normalize gastric dysrhythmias. This has led some investigators to use gastric electrical stimulation to treat patients with gastroparesis. Previous studies and the current state of the field in gastric electrical stimulation in treatment of gastroparesis will be discussed and summarized.     

Chronic intestinal pseudoobstruction as a complication of Duchenne's muscular dystrophy

We report a case of Duchenne's muscular dystrophy complicated by intestinal pseudoobstruction. The patient had recurrent attacks of nausea, vomiting, and abdominal distention for many years, and abdominal films repeatedly showed a dilated and fluid-filled small intestine and colon. Barium studies showed an esophageal diverticulum, reduced esophageal and gastric motility, and a dilated small bowel and colon. Pathologically, the entire gastrointestinal tract had smooth muscle fibrosis, but this was most marked in the esophagus and stomach. We conclude that Duchenne's muscular dystrophy may involve intestinal smooth muscle and produce pseudoobstruction.     

Electrical activity recorded from abdominal surface after gastrectomy or colectomy in humans

The visceral electrical activity recorded from the abdominal surface was studied before and after either total gastrectomy or colectomy. The patterns obtained from fast Fourier transform analysis demonstrated the disappearance of the power peak of approximately 3 cpm after gastrectomy, whereas colectomy did not result in the disappearance of the power peak of approximately 3 and 8-12 cpm. Only the frequencies of approximately 3.5-7.5 cpm were not present after colon surgery. These data demonstrate that the spectral power peaks at frequencies of approximately 3 cpm are entirely related to the stomach because they disappear after gastrectomy; the power peaks between 3.5 and 7.5 cpm are related to the colon because they are present after gastrectomy but not after colectomy; the power peaks between 7.5 and 11 cpm are related to the small intestine because they are present after either gastrectomy or colectomy. The authors conclude that the electrical activity recorded from the abdominal surface and analyzed by fast Fourier transform gives reliable information concerning the electrical activity of the stomach and small intestine, although it is less reliable concerning the electrical activity of the colon.     

Electron microscopic study of neuropeptide Y-containing nerve elements of the guinea pig small intestine

Neuropeptide Y-containing nerve cell bodies and processes were identified by electron microscopic immunocytochemistry in the guinea pig small intestine. Labeled nerve processes were numerous in the myenteric plexus. However, a few immunoreactive nerve fibers were found in all layers of the small intestine. Some of the immunoreactive nerve processes were found in close apposition to the epithelial cells of the crypts of Lieberkühn and to endothelial and smooth muscle cells. The neuropeptide Y-containing nerve cell bodies were preferentially located in the submucous ganglia. In the myenteric plexus many synaptic connections were observed between the neuropeptide Y-immunoreactive nerve fibers and unlabeled nerve cell bodies and other nerve fibers. These findings provide a morphologic basis for the possibility that neuropeptide Y may act as a transmitter and exert postsynaptic effects on intrinsic neurons, in addition to participating in the regulation of smooth muscle activity and epithelial cell functions.     

Electrical activity of the stomach and upper intestine

Thecontrol andresponse electrical activities of the stomach and upper intestine have been described. Control activities are spontaneous, rhythmic activities which do not induce contractions directly but which do control contractions by controlling the appearance of response activities. Properties of the control activity (pacesetter potentials) of the stomach have been described and compared to the properties of the control activity of the upper duodenum. Differences between these control activities were stressed: only in the stomach can single pacesetter potentials be induced locally as desired (by intraarterial acetylcholine) and obliterated locally as desired (by intraarterial catecholamines). Using these procedures, it was shown that the pacesetter potential of the stomach is followed by a refractory period; can be abolished temporarily by collision with a premature potential which spreads in a retrograde direction, or by a local injection of catecholamines proximal to a recording site. These findings and other evidence suggest that the pacesetter potentials of the stomach may differ from those of the intestine, and present a challenge to current models (eg, coupled relaxation oscillators) for the control activities of the gastrointestinal tract. The electrical activity of the stomach can be manipulated experimentally, and this would be advantageous in testing the applicability of models to gastrointestinal electrical activity.Supported by a grant from the Life Insurance Medical Research Fund.     

The arrest of intestinal epithelial `turnover' by the use of x-irradiation

Mitotic activity and hence cell turnover has been abolished by x-irradiation with 1,000 r. in segments of the rat's gut. The whole stomach, the ascending colon, and 5 cm. of jejunum and ileum were irradiated and the changes observed histologically up to three days. Cessation of cell turnover in the small intestine is followed by cell death and desquamation within 24 hours, leading to denudation of the epithelium and collapse of villous structure. The pylorus and colon showed similar but less marked changes. In contrast the fundus and body of the stomach were unaffected by the absence of mitosis over a period of three days. It is concluded that cell turnover is, in part, necessary to maintain the mucosal structure, particularly the villous pattern in the small intestine.     

Coordination of the myoelectrical activity of the large and small intestine

Coordination of the myoelectrical activity of the large and small intestine was studied. Pacemaker cells of intestine are predominantly located at the proximal divisions of large and small intestine and have an increased spontaneous slow-wave activity, which ensures the distribution of excitation in smooth muscle underlying intestines. Due to the ileocecal coordination by sequential motor activity of small and large intestine is provided. The distal direction gradient of slow waves frequency reduction was established. Pacemaker cells possess certain structural specificity and is specialized in the spontaneous bioelectric activity.     

Herbal extracts modulate the amplitude and frequency of slow waves in circular smooth muscle of mouse small intestine

BACKGROUND: Herbal preparations like STW 5 (Iberogast) are widely used drugs in the treatment of dyspepsia and motility-related disorders of the gastrointestinal tract. STW 5 is a phytotherapeutic agent consisting of a fixed mixture of 9 individual plant extracts. The electrophysiological mechanisms of action of STW 5 remain obscure. AIM: The aim of the present study was to investigate whether herbal extracts influence electrophysiological parameters of the small intestine. For this purpose, the resting membrane potential (RMP) and the slow wave rhythmicity of smooth muscle cells of mouse small intestine were observed. METHODS: Intracellular recordings of smooth muscle cells of the circular muscle layer of mouse small intestine were performed using standard microelectrode techniques. After dissection of the mucosa, the small intestine was placed in an organ bath and a microelectrode was applied on a circular smooth muscle cell. The RMP and the amplitude of slow waves were measured in millivolts. RESULTS: The RMP of smooth muscle cells was -59 +/- 1.3 mV. This RMP was significantly depolarized by STW 5 (9.6 +/- 1.6 mV); the depolarizing effects can be mainly attributed to the constituents of matricariae flos, angelicae radix and chelidonii herba. The basal frequency of small intestinal slow waves was 39.5 +/- 1.4 min(-1) and the amplitude was 23.1 +/- 0.9 mV. STW 5 significantly reduced the amplitude and frequency of the slow waves (11.7 +/- 0.8 mV; 33.5 +/- 3.4 min(-1)). This effect on slow waves represents the sum of the effects of the 9 phytoextracts. Whereas angelicae radix and matricariae flos completely blocked slow wave activity, Iberis amara increased the frequency and amplitude, chelidonii herba reduced the frequency and amplitude of the slow waves, mentae piperitae folium reduced the frequency and left amplitude unchanged and liquiritae radix, carvi fructus and melissae folium had no effects. CONCLUSION: Herbal extracts cause changes in smooth muscle RMP and slow wave rhythmicity, up to reversible abolition, by blockade of large conductance Ca2+ channels and other not yet identified mechanisms. In herbal preparations like STW 5 these effects add up to a total effect and this study indicates that herbal preparations which are widely used in dyspepsia and motility-related disorders have characteristic, reproducible, reversible effects on small intestinal electrophysiology.     

The ultrastructural bases for coordination of intestinal motility

The electrical control activity (slow waves) of dog small intestine is characterized by phase locking of potential changes in a frequency plateau in the upper intestine. In the distal intestine, phase locking does not occur, though frequencies of each segment are pulled up (increased) by adjacent, more proximal segments. This suggests poorer coupling in the distal compared to the proximal intestine. Electron microscopic studies of the intestine revealed no differences in appearance or number of nexuses (found only in circular muscle) or of intermediate contacts (found in both muscle layers) in duodenum and upper jejunum as compared with ileum. Thus, differences in cell to cell contacts could not explain poorer coupling in the ileum. No difference in innervation of these two regions was observed. However, evidence was obtained that circular muscle cells of the ileum, unlike those of the duodenum, are not oriented perpendicularly to the longitudinal muscle layer. This could provide a structural basis for poorer coupling and for the observed phase lag of potentials around the circumference of the ileum.     

Blockade of kit signaling induces transdifferentiation of interstitial cells of cajal to a smooth muscle phenotype.

BACKGROUND & AIMS: Interstitial cells of Cajal (ICC) serve as pacemaker cells and mediators of neurotransmission from the enteric nervous system to gastrointestinal muscles. ICC develop from mesenchymal cells that express c-Kit, and signaling via Kit receptors is necessary for normal development of ICC. We studied the fate of functionally developed ICC after blockade of Kit receptors to determine whether ICC undergo cell death or whether the phenotype of the cells is modified. The fate of undeveloped ICC was also investigated. METHODS: Neutralizing, anti-Kit monoclonal antibody (ACK2) was administered to mice for 8 days after birth. ICC in the small intestine were examined by immunohistochemistry and electron microscopy. Occurrence of apoptosis was also assayed. RESULTS: When Kit receptors were blocked, ICC nearly disappeared from the small intestine. Apoptosis was not detected in regions where ICC are normally distributed. Remaining Kit-immunopositive cells in the pacemaker region of the small intestine developed ultrastructural features similar to smooth muscle cells, including prominent filament bundles and expression of the muscle-specific intermediate filament protein, desmin, and smooth muscle myosin. ICC of the deep muscular plexus normally develop after birth in the mouse. Precursors of these cells remained in an undifferentiated state when Kit was blocked. CONCLUSIONS: These data, along with previous studies showing that ICC in the pacemaker region of the small intestine and longitudinal muscle cells develop from the same Kit-immunopositive precursor cells, suggest inherent plasticity between the ICC and smooth muscle cells that is regulated by Kit-dependent cell signaling.     

Insulin and myoelectric activity of the small intestine of the pig

The effect of insulin on the myoelectric activity of the small intestine was determined in conscious pigs. Animals were implanted with electrodes along the small intestine, a strain gage on the stomach and catheters in both saphenous arteries. Feeding modified the migrating myoelectric complex (MMC), a cyclic pattern of action potential activity of the small intestine characteristic of fasting. The first period of regular spiking activity (RSA) on the duodenum after feeding was delayed and was not followed by quiescence. Plasma insulin and glucose concentrations during the first three MMC after feeding were highest just before periods of duodenal RSA. Injection or infusion of insulin into fasted pigs with production of hypoglycemia caused disruption of stomach motility and duodenal electrical activity. The duodenal MMC was not altered when glucose to prevent hypoglycemia was infused together with insulin or when glucose was infused alone. These studies suggest that insulin is not directly responsible for the postprandial modification of MMC activity as insulin infusions only modify the MMC when hypoglycemia occurs.A preliminary report of this work has been published as an abstract (1).     

Kinetics of enteroendocrine cells with implications for their origin: a study of the cholecystokinin and gastrin subpopulations combining tritiated thymidine labelling with immunocytochemistry in the mouse.

Evidence for a common endodermal stem cell has been derived from kinetic studies in mouse small intestine which indicate that the turnover characteristics of endocrine cells are similar to those of other cell lineages (columnar and goblet cells). We have used continuous tritiated thymidine labelling and peptide immunocytochemistry on resin embedded semithin sections, a combination of techniques which have not been used before in the small intestine. Our data show that the turnover time for endocrine cells in the small intestine is 10 days, considerably longer than the four days suggested by previous studies, although for columnar and mucous cell lineages, turnover rates are similar to the published literature. In the stomach, the turnover time was very slow indeed (of the order of 45-60 days). These results show that endocrine cells do not share turnover characteristics with the other cell types and suggest that they constitute a kinetically distinct cell population independent of the other cell lineages. These data are not consistent with a common stem cell origin for gut endocrine cells.     

Interstitial cells of Cajal mediate inhibitory neurotransmission in the stomach.

The structural relationships between interstitial cells of Cajal (ICC), varicose nerve fibers, and smooth muscle cells in the gastrointestinal tract have led to the suggestion that ICC may be involved in or mediate enteric neurotransmission. We characterized the distribution of ICC in the murine stomach and found two distinct classes on the basis of morphology and immunoreactivity to antibodies against c-Kit receptors. ICC with multiple processes formed a network in the myenteric plexus region from corpus to pylorus. Spindle-shaped ICC were found within the circular and longitudinal muscle layers (IC-IM) throughout the stomach. The density of these cells was greatest in the proximal stomach. IC-IM ran along nerve fibers and were closely associated with nerve terminals and adjacent smooth muscle cells. IC-IM failed to develop in mice with mutations in c-kit. Therefore, we used W/W(V) mutants to test whether IC-IM mediate neural inputs in muscles of the gastric fundus. The distribution of inhibitory nerves in the stomachs of c-kit mutants was normal, but NO-dependent inhibitory neuro-regulation was greatly reduced. Smooth muscle tissues of W/W(V) mutants relaxed in response to exogenous sodium nitroprusside, but the membrane potential effects of sodium nitroprusside were attenuated. These data suggest that IC-IM play a critical serial role in NO-dependent neurotransmission: the cellular mechanism(s) responsible for transducing NO into electrical responses may be expressed in IC-IM. Loss of these cells causes loss of electrical responsiveness and greatly reduces responses to nitrergic nerve stimulation.     

Segmental Intestinal Absorption of Ranitidine: Investigative and Therapeutic Implications

The absorption of ranitidine from different segments of the intestinal tract was studied in a subject who had an anatomically divided small intestine. The results indicate that the drug is maximally absorbed from the small bowel rather than the stomach. Subsequent animal studies have shown that absorption of ranitidine in dogs is primarily duodenal followed by jejunal and ileal. The lack of gastric absorption, with predominantly small bowel absorption, may have investigational and therapeutic implications in a variety of gastrointestinal diseases.