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2.
Eur J Pediatr Surg ; 16(6): 407-10, 2006 Dec.
Article in English | MEDLINE | ID: mdl-17211788

ABSTRACT

INTRODUCTION: Dilatation and impaired function of the gut is a condition often seen in newborns with bowel obstruction caused by intestinal atresia. In a previous experimental study in chicken embryos, we established a model to study ultrastructural changes during the development of the enteric nervous system after small bowel ligation. The aim of this study is to investigate the changes of the enteric nervous system (ENS) after gut ligation. METHODS: 56 chicken embryos were investigated. In the operation group fertilized eggs and the allantoic membrane were opened and the small bowel was ligated on embryonal day (ED) 11. The controls were sham-operated. The gut was prepared and harvested for analysis on ED 11, 12, 13, 14, 15, 16, 17 and 18. Silver staining or staining of the specimens for acetylcholinesterase (AchE) was performed. RESULTS: A marked dilatation of the bowel was observed three days after operation (ED 14). The submucosal (PSM) and myenteric plexus (PM) appeared normal at this time, however silver staining showed rarification of the neuronal axonal network between the myenteric and submucosal plexus. Later, on ED 16 an additional rarification of the submucosal plexus was also seen in the operation group using AchE staining, compared to the controls. DISCUSSION: The data suggest that distension of the gut hinders normal development of the ENS in the gut ligation model of chicken embryos. The changes were observed sequentially, starting with rarification of the axonal network between the PM and PSM. Future studies will be required to show whether the changes of the ENS are reversible.


Subject(s)
Intestines/embryology , Intestines/innervation , Myenteric Plexus/embryology , Submucous Plexus/embryology , Submucous Plexus/physiopathology , Animals , Chick Embryo , Dilatation , Intestinal Atresia , Intestines/growth & development , Ligation , Models, Animal , Myenteric Plexus/growth & development , Myenteric Plexus/physiopathology , Submucous Plexus/growth & development
3.
Virchows Arch ; 444(3): 239-46, 2004 Mar.
Article in English | MEDLINE | ID: mdl-14749927

ABSTRACT

An increased number and density of the so-called "giant ganglia" (seven or greater ganglion cells per ganglion) serve as histopathological criteria for a bowel motility disorder called intestinal neuronal dysplasia of the submucous plexus (IND B). However, because these morphological criteria have been defined based upon observations in constipated patients, the diagnostic value of previous studies is open to controversy. Moreover, no age-related reference data from unaffected controls are available. This study reports on data from unaffected controls on the variability of size and distribution of ganglia in the submucous plexus during development. Therefore, for the first time, the normal status has been defined. Four age groups have been defined: (a) premature births, gestational age less than 35 weeks; (b) 1-365 days; (c) 1-14 years and (d) 15 years to greater than 70 years). All of these groups revealed giant ganglia in the submucous plexus. With advancing age, there was a decrease in the number of giant ganglia (from 32.7% in group a to 11.2% in group d) accompanied by an inverse increase in the mean distance between all ganglia (from 0.52 mm in group a to 1.17 mm in group d). The data presented permit the conclusion that the criteria mentioned above are not apt to define IND B as an entity, since they do not allow a sufficient demarcation from the age-correlated normal values presented here.


Subject(s)
Aging , Submucous Plexus/anatomy & histology , Adolescent , Adult , Aged , Child , Child, Preschool , Ganglia/anatomy & histology , Gestational Age , Humans , Infant , Infant, Newborn , Middle Aged , Submucous Plexus/embryology , Submucous Plexus/growth & development
4.
Cell Tissue Res ; 303(2): 301-5, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11291776

ABSTRACT

In the small intestine of both embryonic birds and mammals, neuron precursors aggregrate first at the site of the myenteric plexus, and the submucous plexus develops later. However, in the large intestine of birds, the submucosal region is colonised by neural-crest-derived cells before the myenteric region (Burns and Le Douarin, Development 125:4335-4347, 1998). Using antisera that recognize undifferentiated neural-crest-derived cells (p75NTR) and differentiated neurons (PGP9.5), we examined the colonisation of the murine large intestine by neural-crest-derived cells and the development of the myenteric and submucosal plexuses. At E12.5, when the neural crest cells were migrating through and colonising the hindgut, the hindgut mesenchyme was largely undifferentiated, and a circular muscle layer could not be discerned. Neural-crest-derived cells migrated through, and settled in, the outer half of the mesenchyme. By E14.5, neural-crest-derived cells had colonised the entire hindgut; at this stage the circular muscle layer had started to differentiate. From E14.5 to E16.5, p75NTR- and PGP9.5-positive cells were observed on the serosal side of the circular muscle, in the myenteric region, but not in the submucosal region. Scattered, single neurons were first observed in the submucosal region around E18.5, and groups of neurons forming ganglia were not observed until after birth. The development of the enteric plexuses in the murine large intestine therefore differs from that in the avian large intestine.


Subject(s)
Intestine, Large/innervation , Submucous Plexus/cytology , Submucous Plexus/embryology , Animals , Antigens, Differentiation/analysis , Cell Count , Cell Differentiation/physiology , Cell Movement/physiology , Desmin/analysis , Female , Intestine, Large/embryology , Male , Mice , Mice, Inbred BALB C , Pregnancy , Receptor, Nerve Growth Factor/analysis , Submucous Plexus/chemistry , Ubiquitin Thiolesterase
5.
Arch Pathol Lab Med ; 123(12): 1264-8, 1999 Dec.
Article in English | MEDLINE | ID: mdl-10583933

ABSTRACT

OBJECTIVE: The bcl-2 protein has the functional role of blocking apoptosis, ie, programmed cell death. This protein is widely expressed in the developing central and peripheral nervous systems. The purpose of this study was to map bcl-2 expression in the human enteric nervous system, as this has not previously been done. METHODS: Rectal specimens were obtained at autopsy of 13 fetuses at 13 to 31 weeks of gestation. Normal colon was also obtained from 5 children and 2 adults, and, in addition, ganglionic and aganglionic bowel resected in 11 patients with Hirschsprung disease was examined. Specimens were fixed in formalin, embedded in paraffin, and analyzed with immunohistochemical methods, using antibodies raised against bcl-2 and neuron-specific enolase (NSE). RESULTS: The bcl-2 protein was expressed in myenteric and submucous ganglion cells in fetuses, children, and adults. Nerve fibers of the enteric plexuses that were bcl-2 immunoreactive were few compared with the number of NSE-immunoreactive nerve fibers. In aganglionic bowel no bcl-2-or NSE-immunoreactive ganglion cells were revealed. Results of NSE immunohistochemistry showed clearly stained hypertrophic nerve bundles, known to be of extrinsic origin, which were only weakly bcl-2 immunoreactive. CONCLUSION: Expression of bcl-2 in enteric ganglion cells of the myenteric and submucous plexuses is displayed in the fetus and during childhood and is also retained in adult bowel. Immunohistochemical analysis of bcl-2 provides a good marker for identification of ganglion cells in Hirschsprung disease and may also be valuable for the diagnosis of disorders characterized by hypoganglionosis or hyperganglionosis.


Subject(s)
Enteric Nervous System/metabolism , Hirschsprung Disease/metabolism , Neurons/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Aged , Aged, 80 and over , Child, Preschool , Colon/metabolism , Enteric Nervous System/embryology , Enteric Nervous System/pathology , Fetus/cytology , Fetus/metabolism , Hirschsprung Disease/pathology , Humans , Immunohistochemistry , Infant , Infant, Newborn , Myenteric Plexus/embryology , Myenteric Plexus/metabolism , Myenteric Plexus/pathology , Phosphopyruvate Hydratase/metabolism , Reference Values , Submucous Plexus/embryology , Submucous Plexus/metabolism , Submucous Plexus/pathology
6.
J Comp Neurol ; 414(2): 193-211, 1999 Nov 15.
Article in English | MEDLINE | ID: mdl-10516591

ABSTRACT

The formation of the enteric nervous system (ENS) from neural crest-derived cell precursors requires the growth factor glial cell line-derived neurotrophic factor (GDNF) and the receptors Ret and GDNF family receptor alpha 1 (GFRalpha1). We investigated the location(s), the timing, and the extent to which these GDNF receptors appear in the population of crest-derived precursors that form the avian ENS using immunohistochemistry and in situ hybridization. Sections and whole mounts of embryonic chick gastrointestinal tract were costained with antibodies to the receptors and to HNK-1, a marker for crest-derived cells. Neural crest-derived precursors migrate through the primitive esophagus to colonize the gizzard where an extensive cellular network forms. Ret-immunoreactivity (ir) was found in a network of cells in the gizzard at embryonic day (E)3.5. As development proceeded, Ret-immunoreactive cells appeared at progressively more caudal positions and were present in the colon at E7.5. Costaining with Ret and HNK-1 was performed to determine the number of Ret-immunoreactive cells in the crest-derived population. Ret appeared in some HNK-1 cells in the esophagus and gizzard at embryonic day (E)3.5. During development, the number of crest cells with Ret increased in the ganglia of the gizzard and small intestine. GFRalpha1-ir was also found in HNK-1 cells in the esophagus at E3.5 but did not appear in the gizzard until E4.5. Surprisingly, the colonizing vanguard of crest-derived cells lacked both Ret- and GFRalpha-ir. Between E4.5 and E6.5, the fraction of HNK-1-positive cells expressing GFRalpha1 increased considerably in the foregut. Ret and GFRalpha1 were coexpressed in many cells at E6.5, and the number of such cells increased as development progressed. In the adult, GFRalpha1 and Ret were found in the neuropil of enteric ganglia. We conclude that the population of cells expressing the receptors increases during development and persists in the adult, findings that support a neurotrophic role for GDNF in the formation and maintenance of the avian ENS.


Subject(s)
Chick Embryo/embryology , Drosophila Proteins , Myenteric Plexus/chemistry , Myenteric Plexus/embryology , Nerve Tissue Proteins , Proto-Oncogene Proteins/genetics , Receptor Protein-Tyrosine Kinases/genetics , Submucous Plexus/chemistry , Submucous Plexus/embryology , Age Factors , Animals , Antibodies , Blotting, Western , CD57 Antigens/analysis , Chickens , Duodenum/innervation , ELAV Proteins , Gene Expression Regulation, Developmental , Glial Cell Line-Derived Neurotrophic Factor Receptors , In Situ Hybridization , Neural Crest/chemistry , Neural Crest/embryology , Proto-Oncogene Proteins/analysis , Proto-Oncogene Proteins c-ret , Quail , RNA, Messenger/analysis , RNA-Binding Proteins/analysis , Receptor Protein-Tyrosine Kinases/analysis , Substance P/analysis , Vagus Nerve/chemistry , Vagus Nerve/embryology , Vasoactive Intestinal Peptide/analysis
7.
Anat Rec ; 254(1): 28-38, 1999 01.
Article in English | MEDLINE | ID: mdl-9892415

ABSTRACT

Galanin is a brain-gut peptide that is present in the central and peripheral nervous systems. In the gut, it is contained exclusively in intrinsic and extrinsic nerve supplies, and it is involved overall in the regulation of gut motility. To obtain information about the ontogeny of galanin, we undertook an immunohistochemical study of chicken embryos. The time of first appearance and the distribution patterns of galanin were investigated with fluorescence and streptavidin-biotin-peroxidase (ABC) immunohistochemical protocols by using a galanin polyclonal antiserum. The various regions of the gut and the pancreas were obtained from chicken embryos aged from 3 days of incubation to hatching. All specimens were fixed in buffered picric acid-paraformaldehyde, frozen, and cut with a cryostat. Galanin-immunoreactive neuroblasts were first detected at 4 days in the mesenchyme of the proventriculus/gizzard primordium and within the Remak ganglion. They then extended cranially and caudally, reaching all of the other gut regions at 6.5 days. Galanin-immunoreactive nerve elements mainly occupied the sites of myenteric and submucous plexuses. From day 15, galanin-immunoreactive nerve fibers tended to invade the circular muscular layer and part of the lamina propria of the mucosa. In the pancreas, weak galanin-immunoreactive nerve elements were detected at 5.5 days. They tended to be distributed among the glandular lobules according to the organ differentiation. The widespread distribution during the earlier embryonic stages represents evidence indicating that the neuropeptide galanin may have a role as a differentiating or growth factor. From late embryonic life, its predominant presence in sympathetic nerves and in muscular layers fits with the functions demonstrated previously in adults of other vertebrates for galanin as a modulator of intestinal motility.


Subject(s)
Enteric Nervous System/embryology , Enteric Nervous System/metabolism , Galanin/metabolism , Animals , Chick Embryo , Esophagus/embryology , Esophagus/innervation , Esophagus/metabolism , Gizzard, Avian/embryology , Gizzard, Avian/innervation , Gizzard, Avian/metabolism , Immunohistochemistry , Intestinal Mucosa/metabolism , Intestines/embryology , Intestines/innervation , Myenteric Plexus/embryology , Myenteric Plexus/metabolism , Pancreas/embryology , Pancreas/innervation , Pancreas/metabolism , Proventriculus/embryology , Proventriculus/innervation , Proventriculus/metabolism , Submucous Plexus/embryology , Submucous Plexus/metabolism
8.
J Pediatr Surg ; 31(5): 661-4, 1996 May.
Article in English | MEDLINE | ID: mdl-8861476

ABSTRACT

Classically, development of the human enteric nervous system has been characterized by the early appearance (between 9 and 12 weeks' gestation) of adrenergic and cholinergic nerves. The development of peptidergic innervation occurs much later. Recent studies have indicated that nitric oxide is involved in the nonadrenergic noncholinergic innervation of the gut, mediating its relaxation. The authors have investigated the ontogeny of nitrergic (nitric oxide synthase-containing) neurons of the developing gut. Bowel segments from the esophagus, pylorus, and ileocecal and rectosigmoid regions of 14 fetuses (gestational age range, 12 to 23 weeks) were studied with nicotinamide adenosine dinucleotide phosphate (NADPH) diaphorase histochemistry. By 12 weeks' gestation, nitrergic neurons had appeared in the myenteric ganglia, at all levels of the gut, and had begun plexus formation. Nitrergic innervation in the submucous plexus becomes evident after 14 weeks. As gestational age increases, nitrergic innervation becomes richer and more organized. Increasing numbers of nitrergic nerve fibers are seen in the circular muscle; some of these fibers project from the myenteric plexus. By 23 weeks' gestation, nitrergic innervation has matured to the pattern observed in the postnatal gut. Thus, the onset and pace of development of nitrergic innervation are similar to adrenergic and cholinergic innervation and occur before peptidergic innervation. This study provides morphological evidence of the ontogenetic significance of nitrergic innervation in the human gut and supports previous suggestions that nitric oxide has a pathophysiological role in developmental gut motility disorders.


Subject(s)
Enteric Nervous System/embryology , Nitric Oxide Synthase/physiology , Cell Differentiation/physiology , Digestive System/embryology , Digestive System/innervation , Female , Gastrointestinal Motility/physiology , Gestational Age , Humans , Infant, Newborn , Male , Myenteric Plexus/embryology , NADPH Dehydrogenase/metabolism , Neurons/cytology , Nitric Oxide/physiology , Pregnancy , Submucous Plexus/embryology
9.
J Pediatr Surg ; 22(3): 243-5, 1987 Mar.
Article in English | MEDLINE | ID: mdl-3550029

ABSTRACT

Extensive studies in the chicken embryo have recently supplied more insights into the development of the enteric nervous system, which mainly derives from the vagal neural crest (i.e., the neural crest opposite somites 1 to 7). Crest cells migrate from this region to and via the developing gut. By means of a double labeling technique of both neural crest cells and cells in the S-phase of the cell cycle, we found that these migrating crest cells still proliferate in the gut. Some cells even go through cell division after the formation of a nerve plexus. Some implications for the pathogenesis of congenital innervation abnormalities such as hyperganglionosis and the aganglionosis of Hirschsprung's disease are discussed.


Subject(s)
Digestive System/embryology , Gastrointestinal Motility , Nervous System/embryology , Neural Crest/cytology , Animals , Autoradiography , Cell Division , Cell Movement , Chick Embryo , Digestive System/cytology , Digestive System/innervation , Hirschsprung Disease/embryology , Immunoenzyme Techniques , Myenteric Plexus/embryology , Submucous Plexus/embryology
10.
J Comp Neurol ; 252(4): 493-506, 1986 Oct 22.
Article in English | MEDLINE | ID: mdl-3537021

ABSTRACT

The terminal portion of the ls/ls mouse is congenitally aganglionic because the precursors of enteric neurons fail to enter this region. This animal was studied in order to gain insight into the origin of enteric glia and into the process by which the precursors of these cells colonize the gut. In control (CD-1) mice, immunoreactivity of the glial marker, glial fibrillary acidic protein, appeared for the first time in the fetal bowel at day E16 and, in adults, was much more intense within intraenteric neural elements than in nerves outside the bowel. Glial fibrillary acidic protein developed in tissue cultures of fetal intestine explanted before the protein appeared in situ, and before the bowel became innervated by extrinsic nerves; thus, the precursors of cells able to elaborate glial fibrillary acidic protein must have been present, but unrecognizable, in the original explants. This explant assay demonstrated that these glial precursors were present in all regions of the bowel of control mice, but not in the presumptive aganglionic bowel of ls/ls mice. The nerves (of extrinsic origin) in the aganglionic tissue of ls/ls mice showed a high level of immunoreactive glial fibrillary acidic protein; nevertheless, their ultrastructure was typical of peripheral nerve, not enteric plexus, and they contained Schwann cells, not enteric glia. These observations support the view that enteric glia are derived from the single wave of neural crest colonists that populates the enteric nervous system before the gut receives its extrinsic innervation. These glial precursors, like neuronal precursors, tend to be excluded from the presumptive aganglionic ls/ls bowel. In contrast, Schwann cells grow into the abnormal ls/ls gut with the extrinsic innervation. The enteric microenvironment appears to promote the expression of glial fibrillary acidic protein in both enteric glia and Schwann cells; however, even within the bowel, Schwann cells retain their characteristic morphology. It is thus probable that the normal enteric nervous system contains supporting cells of separate lineages, enteric glia and Schwann cells.


Subject(s)
Glial Fibrillary Acidic Protein/metabolism , Myenteric Plexus/embryology , Neuroglia/cytology , Submucous Plexus/embryology , Animals , Cell Movement , Culture Techniques , Female , Ganglia, Spinal/embryology , Immunoenzyme Techniques , Male , Mice , Mice, Neurologic Mutants , Microscopy, Electron
11.
Anat Embryol (Berl) ; 150(1): 1-6, 1976 Dec 22.
Article in English | MEDLINE | ID: mdl-1015625

ABSTRACT

The chicken intestinal tract from the 6th day of breeding until the 20th day after hatching was examined histochemically for acetylcholinesterase activity. The enzyme was detected first in the perikarya and subsequently also in the ganglion-cells processes. The appearance of the acetylcholinesterase reaction deposit was observed first in the stomach and duodenum and later in the rectum. The histogenetic development of the intramural plexus in the small gut proceeds caudad, but in the colon in the opposite way, i.e., craniad. In the duodenum and colon the nervous development is complete in chicken embryos of 11-12 days and in the rest of the small intestine in embryos of 13-14 days of breeding. In the human embryo of 60 mm CRL the development of the intramural plexus is already completed.


Subject(s)
Autonomic Nervous System/embryology , Digestive System/innervation , Acetylcholinesterase/analysis , Animals , Chick Embryo , Colon/innervation , Duodenum/innervation , Humans , Myenteric Plexus/embryology , Rectum/innervation , Stomach/innervation , Submucous Plexus/embryology
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