Interstitial cells of Cajal (ICC) are the pacemalkers in gastrointestinal muscles, and these cells also mediate or transduce inputs from the enteric nervoius system. Immunolabelling of interstitial cells of ICC in intestinal wall is recently developed by using specific marker, anti-c-kit antibody. Immunohistochemistry was done for c-Kit-positive ICC network in attempt to provide a morphological basis for the mechanism regulating gastro-intestinal movement. Cryosection and whole-mount preparations of mouse ileum and colon were immunolabelled using the anti-c-Kit. Immunolabelled specimens were observed under a confocal laser scanning microscopy. According to three dimensional reconstruction study, it was found that the c-Kit-positive cells were widely distributed in the intestinal wall: (1) circular muscle layer, (2) myenteric plexus, (3) deep muscular plexus in ileum, (4) submucosal plexus and longitudinal muscle layer in colon. The characteristic profiles of ICC containing c-Kit-positive cells provide a morphological basis upon the mechanism regulating gastro-intestinal motility.
Animals ; Colon ; Ileum ; Immunohistochemistry ; Interstitial Cells of Cajal* ; Intestines* ; Mice* ; Microscopy, Confocal ; Muscles ; Myenteric Plexus ; Submucous Plexus

Hirschsprung's disease (HD) is one of the major pediatric gastrointestinal disease entities which is associated with an absence or lack of intrinsic ganglion cells in the myenteric and submucosal plexus in the gastrointestinal tract. It is commonly assumed to be a sex-modified multifactorial trait. The development of diagnostic and therapeutic modalities has been ongoing. Herein, we experienced two siblings who were confirmed as having HD histologically and were treated. We think further family evaluation regarding HD is needed. Also we could see a changing modality of diagnosis and treatment.
Ganglion Cysts ; Gastrointestinal Diseases ; Gastrointestinal Tract ; Hirschsprung Disease ; Humans ; Siblings ; Submucous Plexus

BACKGROUND: This study was done to obtain comprehensive data on changes in the structural components of the enteric nervous system in pediatric patients with intestinal pseudo-obstruction (IPO). We evaluated routinely processed, in formalin-fixed tissues by quantitative morphometric analysis. In addition, we used formalin-fixed tissue to explore the possibility of using previously proposed diagnostic criteria to evaluate frozen serial sections for intestinal neuronal dysplasia (IND) type B and hypoganglionosis. METHODS: We analyzed data for 19 IPO cases. Morphometric analysis for quantification of ganglia and ganglion cells (GCs) was done for the myentric and the submucous plexus. In addition, we determined the presence of immature GCs and the distribution of nerve fibers and interstitial cells of Cajal (ICC). RESULTS: Nine patients showed combined hypoganglionosis, IND, and decreased ICC; others showed various combinations of these. Several morphometric factors were significantly different between patient groups as well as being different than the control group. CONCLUSIONS: Our pediatric IPO cases showed extensive overlapping of pathological findings. And the findings suggest the utility of using previously proposed morphometrically measured factors in multiple frozen sections as diagnostic criteria for IND type B and hypoganglionosis in formalin-fixed tissue.
Enteric Nervous System ; Frozen Sections ; Ganglia ; Ganglion Cysts ; Humans ; Interstitial Cells of Cajal ; Intestinal Pseudo-Obstruction ; Nerve Fibers ; Neurons ; Submucous Plexus

Bowel obstruction is a rare complication of pregnancy. The usual causes include previous abdominal surgery, volvulus, intussusception, colonic neoplasm, or the enlarging uterus. Bowel obstruction secondary to uncorrected Hirschsprung's disease as a complication of pregnancy is difficult to diagnosis, its occurrence can have grave implications for both mother and fetus, and anticipation of dystocia. Hirschsprung's disease is diagnosed and treated in the neonatal period. Persistence of Hirschsprung's disease into adulthood is very rare and confirmed by rectal biopsy providing the absence of the ganglion cell in Auerbach and Meissner's plexus. We experienced vaginal Delivery in Hirschsprung's disease complicating pregnancy and report our own case study with a brief literature review.
Biopsy ; Colonic Neoplasms ; Diagnosis ; Dystocia ; Female ; Fetus ; Ganglion Cysts ; Hirschsprung Disease* ; Humans ; Intestinal Volvulus ; Intussusception ; Mothers ; Pregnancy* ; Submucous Plexus ; Uterus

This study was performed to investigate the morphometric and ultrastructural change in the adult and aged rat small intestine. The myenteric and submucous plexuses were stained by NADH-TR in the ileum of adult Sprague-Dawley rats (3 mo., 300~350 gm) and aged rats (24 mo., 500~550 gm). The neurons of myenteric and sumucous plexuses were divided into 3 groups depending on their cell body morphology. Type 1 cells were polygonal or round with abundant cytoplasm. Type 2 cells were spindle shaped and type 3 cells were small and round with scanty cytoplasm. The nerve cell numbers and sizes were measured using an image analyzer (VIDAS, Carl Zeiss, Co., Ltd.). Ultrastructural changes were observed by JEM-1200 EXII (JEOL Co., Ltd.) electron microscope. The result obtained are as followed: 1. In adult rats, majority of neuron population were type 3 and neuron density (total numbers/1 mm2) was more higher in submucous plexus than in the myenteric plexus. 2. Statistically significant loss of type 1 and type 2 neurons were in myenteric and submucous plexus of aged rat small intestine. 3. All types of neuron sizes were increased in aged myenteric and submucous plexuses. 4. Lipofusin granules were prominent in the cytoplasm of aged rat. Cell organelles were not shown degenerative change. These results suggest that type 1 and type 2 nerve cells which is originated from autonomic nerves were lost in aged rat small intestine. Ultrastructurally lipofusin granules were prominent in the cytoplasm of aged rat and the cell organelles were not degenerated.
Adult* ; Aging ; Animals ; Autonomic Pathways ; Cytoplasm ; Enteric Nervous System* ; Humans ; Ileum ; Intestine, Small* ; Myenteric Plexus ; Neurons ; Organelles ; Rats* ; Rats, Sprague-Dawley ; Submucous Plexus

Hirschsprung's disease is a disorder caused by the absence of ganglion cells in the colon and rectum. It has an incidence of 1 in 5000 births, the majority diagnosed and treated in the neonatal period due to symptoms of intestinal obstruction. Persistence of Hirschsprung's disease into adulthood is very rare. In such patients, prolonged periods of constipation are a common problem. For the diagnosis, a colon study and anorectal manometry are performed, and the presence of the disease is confirmed by an excisional biopsy proving the absence of the ganglion cell in Auerbach and Meissner's plexus. Although various surgical procedures have been performed, there is no obvious optimal choice for treatment of Hirschsprung's disease in adolescents and adults. We experienced two cases of Hirschsprung's disease, confirmed by a rectal biopsy, in 20-year patients. Prior to a definitive operation, a sigmoid loop colostomy was performed due to severe dilatation of the left colon and rectum. Six months later, one patient was treated using Duhamel's procedure, and the other by using a proctosigmoidectomy and coloanal anastomosis. No postoperative complications were observed, and the patients had bowel movements three to four times a day. Despite its infrequent incidence, adult Hirschsprung's disease should be suspected in patients who have had lifelong constipation. Several successful surgical treatments have been used for treatment of patients with adult Hirschsprung's disease. In our cases, the functional results of Duhamels' procedure and of a proctosigmoidectomy with coloanal anastomosis were satisfactory.
Adolescent ; Adult* ; Biopsy ; Colon ; Colon, Sigmoid ; Colostomy ; Constipation ; Diagnosis ; Dilatation ; Ganglion Cysts ; Hirschsprung Disease* ; Humans ; Incidence ; Intestinal Obstruction ; Manometry ; Parturition ; Postoperative Complications ; Rectum ; Submucous Plexus

BACKGROUND/AIMS: Physical and/or emotional stresses are important factors in the exacerbation of symptoms in irritable bowel syndrome (IBS). Several lines of evidence support that a major impact of stress on the gastrointestinal tract occurs via the enteric nervous system. We aimed to evaluate histological changes in the submucosal plexus (SMP) and myenteric plexus (MP) of the distal ileum in concert with the intestinal motor function in a rat model of IBS with diarrhea. METHODS: The rat model was induced by heterotypic chronic and acute stress (CAS). The intestinal transit was measured by administering powdered carbon by gastric gavage. Double immunohistochemical fluorescence staining with whole-mount preparations of SMP and MP of enteric nervous system was used to assess changes in expression of choline acetyltransferase, vasoactive intestinal peptide, or nitric oxide synthase in relation to the pan neuronal marker, anti-Hu. RESULTS: The intestinal transit ratio increased significantly from control values of 50.8% to 60.6% in the CAS group. The numbers of enteric ganglia and neurons in the SMP were increased in the CAS group. The proportions of choline acetyltransferase- and vasoactive intestinal peptide-immunoreactive neurons in the SMP were increased (82.1 ± 4.3% vs. 76.0 ± 5.0%, P = 0.021; 40.5 ± 5.9% vs 28.9 ± 3.7%, P = 0.001), while nitric oxide synthase-immunoreactive neurons in the MP were decreased compared with controls (23.3 ± 4.5% vs 32.4 ± 4.5%, P = 0.002). CONCLUSIONS: These morphological changes in enteric neurons to CAS might contribute to the dysfunction in motility and secretion in IBS with diarrhea.
Animals ; Carbon ; Choline ; Choline O-Acetyltransferase ; Diarrhea* ; Enteric Nervous System ; Fluorescence ; Ganglia ; Gastrointestinal Motility ; Gastrointestinal Tract ; Ileum ; Intestine, Small* ; Irritable Bowel Syndrome* ; Models, Animal* ; Myenteric Plexus ; Neurons* ; Nitric Oxide ; Nitric Oxide Synthase ; Rats* ; Stress, Psychological ; Submucous Plexus ; Vasoactive Intestinal Peptide

BACKGROUND/AIMS: Digestion of dietary protein elevates intraluminal concentrations of glutamate in the small intestine, some of which gain access to the enteric nervous system (ENS). Glutamate, in the central nervous system (CNS), is an excitatory neurotransmitter. A dogma that glutamatergic neurophysiology in the ENS recapitulates CNS glutamatergic function persists. We reassessed the premise that glutamatergic signaling in the ENS recapitulates its neurotransmitter role in the CNS. METHODS: Pharmacological analysis of actions of receptor agonists and antagonists in concert with immunohistochemical localization of glutamate transporters and receptors was used. Analysis focused on intracellularly-recorded electrical and synaptic behavior of ENS neurons, on stimulation of mucosal secretion by secretomotor neurons in the submucosal plexus and on muscle contractile behavior mediated by musculomotor neurons in the myenteric plexus. RESULTS: Immunoreactivity for glutamate was expressed in ENS neurons. ENS neurons expressed immunoreactivity for the EAAC-1 glutamate transporter. Neither L-glutamate nor glutamatergic receptor agonists had excitatory actions on ENS neurons. Metabotropic glutamatergic receptor agonists did not directly stimulate neurogenic mucosal chloride secretion. Neither L-glutamate nor the metabotropic glutamatergic receptor agonist, aminocyclopentane-1,3-dicarboxylic acid (ACPD), changed the mean amplitude of spontaneously occurring contractions in circular or longitudinal strips of intestinal wall from either guinea pig or human small intestinal preparations. CONCLUSIONS: Early discoveries, for excitatory glutamatergic neurotransmission in the CNS, inspired enthusiasm that investigation in the ENS would yield discoveries recapitulating the CNS glutamatergic story. We found this not to be the case.
Amino Acid Transport System X-AG ; Animals ; Central Nervous System ; Dietary Proteins ; Digestion ; Enteric Nervous System* ; Glutamic Acid* ; Guinea Pigs ; Humans ; Intestine, Small ; Intestines ; Muscles ; Myenteric Plexus ; Neurons ; Neurophysiology ; Neurotransmitter Agents ; Proteolysis ; Receptors, Glutamate ; Submucous Plexus ; Synaptic Transmission

Recently, it has been postulated that diabetic autonomic neuropathy is caused by reduction in availability of nerve growth factor (NGF) in enteric nervous system. This experiments were performed to determine the changes of the distribution of enteric neuropeptide by diabetes and these changes could be prevented by administration of NGF. Sprague Dawley rats (200~250gm) were made diabetic by a single intraperitoneal injection of streptozotocin 65 mg/kg in saline. Recombinant human NGF (Sigma, Co., Ltd.) were administered at a dose of 500ng/kg subcutaneously every day for consecutive 4 weeks after streptozotocin administration. After 4 weeks, rats were anesthetized with ether and perfused with 4% paraformaldehyde. ileum was dissected and prepared by whole mount preparation method. Prepared segments were immunostained for substance p, calcitonin gene-related peptide, vasoactive intestinal peptide, and galanin by PAP technique. For the observation of the interstitial cells of Cajal, segments were immersed in Champy-Maillet solution for 2 days Results obtained were as follows: 1. In myenteric plexus of diabetic rats, substance P-like and VIP-like immunoreactivity were not changed compared with that of the control group. CGRP-like and galanin-like immunoreactivity were decreased in diabetic group and immunoreactive cells for CGRP and galanin were also decreased 18.1% (P<0.01) and 43.7% (P<0.01) respectively. 2. In NGF administerd diabetic group, immunoreactivity of substance p, VIP, galanin in myenteric plexus were slightly increased and immunoreactive cells for substancre p, VIP, galanin were almost the same as that of the control group. However, immunoreactive cells for CGRP of myenteric plexus were not changed by NGF. 3. In submucous plexus of diabetic rats, immunoreactivity of all four neuropeptides(substance p, CGRP, VIP, galanin) were decreased compared with that of the control group. Immunoreactive cells for substance p, CGRP, VIP, and galanin were also decreased in 38.8%, 77.6%, 33.0%, and 35.7%, respectively (P<0.01). 4. In NGF administered diabetic group, immunoreactivities of substance p, VIP and galanin in submucous plexus were increased and the immunoreactive cells were increased significantly compared to diabetic group. However, immunoreactive cells for CGRP of submucous plexus were not changed by NGF. 5. Interstitial cells of Cajal of diabetic group were decreased 7.4% ovoidal cells (A type) and 28.3% round cells (B type) In NGF administered group, the morphology and the number of ICC were not different to the control group. With the above results, it could be assumed that NGF prevent the damage of neurotransmitter and ICC in enteric nervous system.
Animals ; Calcitonin Gene-Related Peptide ; Diabetic Neuropathies ; Enteric Nervous System ; Ether ; Galanin ; Humans ; Ileum ; Injections, Intraperitoneal ; Interstitial Cells of Cajal ; Myenteric Plexus ; Nerve Growth Factor* ; Neuropeptides ; Neurotransmitter Agents ; Peristalsis* ; Rats* ; Rats, Sprague-Dawley ; Streptozocin ; Submucous Plexus ; Substance P ; Vasoactive Intestinal Peptide

PURPOSE: Intestinal neuronal dysplasia (IND) causes intestinal pseudo-obstruction and shares clinical features with Hirschsprung's disease. Diagnosis of IND involves histopathological features of an intestinal biopsy, but diagnostic criteria are controversial and optimal treatment is unclear. We determined the pathological findings for diagnosing IND in infants and the significance of surgical treatment. METHODS: We retrospectively studied 4 patients who received bowel surgery for an intestinal obstruction without a definite obstructive cause that were subsequently diagnosed as IND by postoperative pathology. The clinical history and results of immunohistochemistry for ganglion and nerve fibers (NCAM, NSE, cathepsin D, synaptophysin) were compared between patients and control cases. RESULTS: All 4 patients were premature babies with symptoms of poor oral intake and abdominal distention. Surgical treatment was segmental resection of the small bowel in one case, segmental resection of the small bowel and double-barreled ileostomy in one case with NEC, and a temporary ileostomy for decompression and appendectomy for biopsy in 2 cases. The first 2 patients died of sepsis and DIC, respectively. The postoperative course of the other 2 patients was excellent for long-term follow up (30+/-6months). Patients with IND showed significantly more submucosal giant plexuses and ganglia in the submucosal plexus, a higher percentage of giant plexus in the 20 submucosal plexus, as well as increased incidence of heterotopic ganglia in the lamina propria, bud-like ganglia, anisomorphic ganglia, and immature ganglia. CONCLUSION: Proper surgical treatment of persistent intestinal pseudo-obstruction, including IND, can affect the prognosis and recovery of bowel function, with positive pathological findings helpful for diagnosing IND in infancy.
Appendectomy ; Biopsy ; Cathepsin D ; Dacarbazine ; Decompression ; Follow-Up Studies ; Ganglia ; Ganglion Cysts ; Hirschsprung Disease ; Humans ; Ileostomy ; Immunohistochemistry ; Incidence ; Infant ; Intestinal Obstruction ; Intestinal Pseudo-Obstruction ; Mucous Membrane ; Nerve Fibers ; Neurons ; Prognosis ; Retrospective Studies ; Sepsis ; Submucous Plexus