Krammer Hj

Organization of the enteric nervous system in the human colon demonstrated by wholemount immunohistochemistry with special reference to the submucous plexus

To demonstrate the normal topography and structure of the enteric nervous system (ENS) in the human colon, the colonic wall of patients (n = 10, mean age 66.3 years), who underwent abdominal surgery unrelated to intestinal motility disorders, was submitted to wholemount immunohistochemistry. The specimens were stretched out and separated into the tunica muscularis, the outer and inner portion of the tela submucosa and the tunica mucosa. Prior to the application of the neuronal marker Protein Gene Product (PGP) 9.5, the laminar preparations were pretreated with the maceration agent KOH. The plexus myentericus was composed of prominent ganglia and interconnecting nerve fiber strands (NFS) forming a polygonal network, which was denser in the descending than in the ascending colon. Nerve cells were observed within the ganglia as well as in primary, secondary and tertiary NFS. The latter ramified into the adjacent smooth muscle layers, which contained the aganglionated plexus muscularis longitudinalis and circularis. The submucous plexus comprised three nerve networks of different topography and architecture: the delicate plexus submucosus extremus consisted of parallel orientated NFS with isolated nerve cells and small ganglia and was located at the outermost border of the tela submucosa adjacent to the circular muscle layer. The plexus submucosus externus was closely associated with the plexus submucosus extremus and composed of larger ganglia and thicker NFS. The plexus submucosus internus was situated adjacent to the lamina muscularis mucosae and formed a network with denser meshes but smaller ganglia and NFS than the plexus submucosus externus. The NFS of the aganglionated plexus muscularis mucosae followed the course of the smooth muscle cells of the lamina muscularis mucosae. The honeycomb-like network of the plexus mucosus was located within the lamina propria mucosae and divided into a subglandular and a periglandular portion. Single and accumulated nerve cells were observed within the plexus mucosus as a regular feature. The findings confirm the complex structural organisation of the ENS encountered in larger mammals, in particular the subdivision of the submucous plexus into three different compartments. PGP 9.5-immunohistochemistry applied to wholemount preparations comprehensively visualized the architecture of the intramural nerve plexus in human colonic specimens. In addition to conventional cross-sections, this technique allows a subtle assessment and classification of structural alterations of the ENS in patients with colorectal motor disorders.

Immunohistochemical visualization of the enteric nervous system using antibodies against Protein gene product (PGP) 9.5

Both the investigation of the enteric nervous system and the diagnosis of its pathological changes require reliable staining methods. In order to assess the potential of protein gene product (PGP) 9.5 as a marker for the enteric nervous system, we examined its immunoreaction in whole-mount preparations of the guinea pig and porcine small intestine, using a rabbit polyclonal antiserum. The immunohistochemical technique reveals the fundamental architectural features of the ganglionic and aganglionic plexuses. Furthermore, it enables a reproducible and differentiated visualization of the enteric nerve cells to be made, so that the various nerve cell types can be morphologically identified.

Topography of the enteric nervous system in Peyer's patches of the porcine small intestine

The mechanisms of intercommunication between the immune and nervous systems are not fully understood. In the case of the intestine, the enteric nervous system is involved in the regulation of immune responses. It was therefore decided to employ immunohistochemical techniques to investigate the structural organization of the enteric nervous system in Peyers patches of the porcine small intestine. Using antibodies against various nervous system-specific markers (protein gene product 9.5, neuron-specific enolase, neurofilament 200, S-100 protein and the glial fibrillary acidic protein), an intimate and specific structural association could be demonstrated between enteric nerves and the compartments of Peyers patches: follicles, interfollicular regions and domes. Peyers patches have a close topographical relationship to the two submucosal plexuses. Enteric nerves are located around the follicle in the interfollicular area — the so-called “traffic area”-and in the dome area, which plays an important role in the uptake and presentation of antigens.

Abnormalities of the enteric nervous system in heterozygous endothelin B receptor deficient (spotting lethal) rats resembling intestinal neuronal dysplasia

Background: A homozygous mutation of the endothelin B receptor (EDNRB) gene in spotting lethal (sl/sl) rats leads to Hirschsprungs disease (HSCR) with long segmented aganglionosis. However, the effects on the development of the enteric nervous system (ENS) promoted by a heterozygous mutation of the EDNRB gene are not known. The present study aimed to describe and morphometrically assess the phenotypic abnormalities of the ENS in heterozygous (+/sl) EDNRB deficient rats in comparison with homozygous (sl/sl) EDNRB deficient and wild-type (+/+) rats. Methods: The distal small intestine, caecum, and colon were obtained from sl/sl, +/sl, and +/+ rats. To demonstrate the three dimensional organisation of the ENS, the intestinal wall was microdissected into wholemounts and incubated against the pan-neuronal marker protein gene product 9.5. Assessment of the ENS included morphometric quantification of ganglionic size and density, the number of nerve cells per ganglia, and the diameter of nerve fibre strands within both the myenteric and submucous plexus. Results: Sl/sl rats were characterised by complete aganglionosis resembling the same histopathological features observed in patients with HSCR. +/sl rats revealed more subtle abnormalities of the ENS: the submucous plexus was characterised by a significantly increased ganglionic size and density, and the presence of hypertrophied nerve fibre strands. Morphometric evaluation of the myenteric plexus did not show statistically significant differences between +/sl and +/+ rats. Conclusions: In contrast with sl/sl rats, +/sl rats display non-aganglionated malformations of the ENS. Interestingly, these innervational abnormalities resemble the histopathological criteria for intestinal neuronal dysplasia (IND). Although IND has been described in several intestinal motility disorders, the concept of a clearly defined clinical-histopathological entity is still controversially discussed. The present findings support the concept of IND based on clearly defined morphological criteria suggesting a genetic link, and thus may provide a model for human IND. Furthermore, the data underline the critical role of the “gene dose” for the phenotypic effects promoted by the EDNRB/EDN3 system and confirm that the development of the ENS is not an “all or none” phenomenon.

Structural differences of the enteric nervous system in the cattle forestomach revealed by whole mount immunohistochemistry

The specific motility patterns of the forestomach of ruminants, composed of three structurally distinct compartments (rumen, reticulum, omasum), require an elaborate intramural innervation. To demonstrate the complex structure of the enteric nervous system (ENS), whole mount preparations obtained from different sites of the bovine forestomach were submitted to immunohistochemical procedures in which neuronal (protein gene product 9.5, neurofilament 200) and glial (protein S-100, glial fibrillary acid protein) markers were applied. Immunohistochemistry performed on whole mounts allowed a detailed two-dimensional assessment of the architecture of the intramural nerve networks. Generally, the myenteric and submucosal plexus layers were composed of ganglia and interconnecting nerve fiber strands, whereas the mucosal plexus consisted of an aganglionated nerve network. However, the texture of the ENS showed considerable regional differences concerning the ganglionic size, shape and density and the arrangement of nerve fiber strands. The myenteric plexus of the ruminal wall, showing a low ganglionic density and wide polygonal meshes, contrasted with the nerve network within the ruminal pillar which consisted of ropeladder-like nerve fiber strands and parallel orientated ganglia. The highest ganglionic density was observed at the reticular groove, the most prominent ganglia were found within the omasal wall. Branches of the vagal nerve frequently ramified within the myenteric plexus layers. The submucosal plexus of the rumen was divided into an external and internal layer; the reticular submucosal plexus followed the cristae and cellulae reticuli, the omasal submucosal (sublaminar) plexus showed intra- and parafascicular ganglia apart from ganglia located at the junctions of the nerve network. The mucosal plexus of the rumen consisted of thin nerve fascicles ramifying between the ruminal papillae, and reticular mucosal nerve fibers passed throughout the base of the cellulae reticuli. The highly specialised nerve network of the intralaminar omasal plexus showed radial and transverse trajectories reflecting the spatial arrangement of the intralaminar musculature. The demonstrated structural complexity of the ENS reflects the functional complexity of the ruminant forestomach and indicates the relatively high degree of autonomy in coordinating the different motility patterns required for the processing of the ingesta.

Immunohistochemical demonstration of nerves and nerve cells in human and porcine ureters

There are divergent opinions about the origin of ureteral motility. It is either a myogenic or a neutral phenomenon. Furthermore, the existence of nerve cells in the ureter is also a controversial question. In the present study we applied immunohistochemical methods to human and porcine ureters in an attempt to clarify the mattern. As neuronal markers we used anti-PGP 9.5 and anti-NSE, and as a glial marker anti-S-100. In the whole mount preparations of pig ureter we observed two neuronal plexuses on both sides of the tunica muscularis. The inner plexus consisted of both nerve bundles and nerve cells, whereas the outer one did not contain any nerve cells. In the human ureter we found a ganglion with nerve cells beneath the tunica muscularis and the tunica adventitia.

The enteric nervous system and interstitial cells of Cajal. Changes in chronic constipation in adults

ZusammenfassungIntestinale Innervationsstörungen gehören zu den vielfältigen Ursachen der chronischen Obstipation und müssen in der differenzialdiagnostischen Abklärung besonders berücksichtigt werden. Das enterische Nervensystem ist als eigenständiges, größtes peripheres Nervensystem („brain in the gut“) maßgeblich an der Vermittlung intestinaler Motilität beteiligt. Morphologisch lassen sich unterschiedliche Nervenzelltypen beschreiben, die sich zu intramuralen Nervenplexus gruppieren und ein breites Spektrum von Transmittern ausschütten. Fehlbildungen oder Läsionen des enterischen Nervensystems können zu teilweise schwerwiegenden intestinalen Passagestörungen führen, die sich klinisch als therapieresistente Obstipation äußern. Im Gegensatz zur frühen Manifestation der Aganglionose bleiben erworbene oder nichtaganglionäre Veränderungen der Nervenplexus oft bis ins Erwachsenenalter unerkannt. Die histopathologische Diagnostik erfolgt über die enzym- oder immunhistochemische Darstellung der Nervengeflechte an Schnittpräparaten oder Schichtpräparaten, die eine flächenhafte Beurteilung der Nervengeflechte erlauben. Um Hypoganglionosen, enterische Ganglionitiden oder Veränderungen der interstitiellen Cajal-Zellen diagnostizieren zu können, sind Ganzwandbiopsien erforderlich. Interstitielle Cajal-Zellen generieren die „Slow-wave-Aktivität“. Die Dichte der intramuskulären Cajal-Zell-Netzwerke ist bei erwachsenen Patienten mit primärer chronischer Obstipation und Megakolon deutlich vermindert.AbstractIntestinal innervation disorders are part of the broad etiological spectrum of chronic constipation and need to be specifically addressed in differential diagnosis. The enteric nervous system constitutes the largest peripheral nervous system of its own (“brain in the gut”), and is involved in the mediation of intestinal motility. Morphologically different nerve cell types aggregate into intramural plexus layers and release a multitude of neurotransmitters. Malformations or lesions of the enteric nervous system may lead to a severely prolonged intestinal transit time resulting in chronic constipation resistant to conservative treatment. In contrast to the early manifestation of aganglionosis, non-aganglionic or acquired alterations to the intramural nerve plexus often remain unrecognised up to adulthood. Histopathological diagnosis is carried out by enzyme or immunohistochemical staining, either on sections or whole mount preparations, allowing an optimal visualization of the nerve plexus architecture. To diagnose hypoganglionosis, enteric ganglionitis or alterations in interstitial cells of Cajal, full-thickness biopsies are required. Interstitial cells of Cajal contribute significantly to the mediation of intestinal motility by generating “slow wave” activity. In adult patients with slow-transit constipation and megacolon, the intramuscular networks of the interstitial cells of Cajal show a significantly reduced density.

Ist die Divertikelkrankheit mit intestinalen Innervationsstörungen assoziiert

Neben ballaststoffarmer Ernahrung und Bindegewebsveranderungen der Darmwand gelten intestinale Motilitatsstorungen als Ursache der Divertikelkrankheit (DK). Da die Vermittlung intestinaler Motilitat masgeblich dem enterischen Nervensystem unterliegt, wurden die intramuralen Nervengeflechte von Patienten mit DK und einer Kontrollgruppe untersucht. Die Beurteilung der immunhistochemisch (PGP 9,5) dargestellten Nervenplexus erfolgte an Schnitt- und Schichtpraparaten der Darmwand. Bei 25% der Patienten mit DK fand sich eine oligoneuronale Hypoganglionose des Plexus myentericus. 41,6% zeigten hypertrophe submukose Nervenfaserstrange auf, 16,6% zusatzlich Riesenganglien. Die Befunde belegen, dass die DK mit intestinalen Innervationsstorungen assoziiert ist. Daher sollten insbesondere bei Divertikeltragern mit obstipativer Symptomatik und einem hyperkontraktilen Kolon Fehlbildungen des enterischen Nervensystems atiologisch berucksichtigt werden.

GFAP-Immunreaktivität in der Neomukosa des Ileums auf einem Serosa Patch des Colons beim Hund

Summary A Short bowel was induced in 8 beagle bitches by resecting 85 % of the small intestine. Following resection, a tangential fixation was performed in antiperistaltic direction between the ileum and the proximal colon. A 15 cm segment of the remaining ileum was incised opposite to its mesenterial attachment and sewn onto the serosa of the neighboring segment of colon (Serosa patching). After 20 weeks of observation, segments of the ileum, the colon and the anastomosed intestinal segment were removed. The neomucosa covering the patch surface was investigated using immunohistoche-mical methods in order to demonstrate the presence of nerve fibers. The immunofluorescence reaction using anti-GFAP (glial fibrillary acidic protein) clearly demonstrated immunoreactivity in the formed neomucosa and in the underlying granulation tissue. The immuno-reactive fibers were comparable with fibers demonstrated in the original mucosa. This finding is an indication that the neomucosa of a serosa patch may be functionally intact because the presence of nerves suggests that mucosal functions, e.g. resorption and secretion, are under nervous regulation.

Leitlinie: Anale Feigwarzen

Feigwarzen (Condylomata acuminata, spitze Kondylome) sind gutartige epitheliale Tumoren mit verschiedenen Ausprägungsformen. Je nach Lokalisation können sie als stecknadelkopfgroße, meist weißlich, braun oder rötlich pigmentierte Papeln sowohl solitär als auch multipel, beetartig konfluierend, aber auch als große blumenkohlartige Tumoren (destruierend: Buschke-Löwenstein-Tumor, nicht destruierend: Condylomata gigantea) imponieren.