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  Table of Contents    
ORIGINAL ARTICLE  
Year : 2011  |  Volume : 54  |  Issue : 2  |  Page : 269-272
Migration and maturation pattern of fetal enteric ganglia: A study of 16 cases


1 Department of Pathology, Burdwan Medical College, Kolkata, India
2 Department of Pathology, Institute of Post Graduate Medical Education and Research, Kolkata, India
3 Department of Gastroenterology, Institute of Post Graduate Medical Education and Research, Kolkata, India
4 Park Children's Center for Treatment and Research, Kolkata, India

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Date of Web Publication27-May-2011
 

   Abstract 

Aims: To study the migration and developmental pattern of ganglion cells in fetuses aged 9-21 weeks, and to document whether the migration was occurring circumferentially equally in the entire axis or if there were discrepancies in different portions at the same level. Settings and Design: The hypothesis regarding the pathogenesis of Hirschsprung's disease mainly revolves around two schools. One is the single gradient migration of ganglia and the other is a dual gradient migration theory. Understanding the embryological development of enteric ganglia is necessary to study the pathogenesis of intestinal innervation disorders. Materials and Methods: We studied the development of intestinal ganglia in fetuses aged 9-21 weeks. Serial longitudinal sections from the colon were studied, the first one including the squamo-columnar junction, for the presence and the nature of ganglion cells with Hematoxylin and Eosin, and neurone-specific enolase immunostaining. Transverse sections from proximal gut were studied in a similar fashion. Thus, we evaluated the migration pattern as well as the nature of ganglia in the fetuses. We also measured the length of distal aganglionic segment in these growing fetuses. Results: We noted that ganglion cells appear first in the myenteric plexus followed by deep and superficial submucous plexus. We also found evidences in favor of dual migration theory, and the distal aganglionic segment varies around the circumference of the rectal wall. Conclusions: We got evidences in support of a dual migration pattern of intestinal ganglion cells. The level of distal aganglionic segments when measured from squamo-columnar junction varied with the age of gestation and the length was incongruous. The description of distal aganglionic segment may help surgeons while taking biopsies or during operative procedures.

Keywords: Fetus, ganglia, intestinal neuronal migration, neurone-specific enolase

How to cite this article:
Bandyopadhyay R, Chatterjee U, Bandyopadhyay SK, Basu AK. Migration and maturation pattern of fetal enteric ganglia: A study of 16 cases. Indian J Pathol Microbiol 2011;54:269-72

How to cite this URL:
Bandyopadhyay R, Chatterjee U, Bandyopadhyay SK, Basu AK. Migration and maturation pattern of fetal enteric ganglia: A study of 16 cases. Indian J Pathol Microbiol [serial online] 2011 [cited 2019 Dec 12];54:269-72. Available from: http://www.ijpmonline.org/text.asp?2011/54/2/269/81589



   Introduction Top


An understanding of the normal embryological development of the enteric nervous system is necessary to study the intestinal neuronal disorders. There are different hypotheses regarding the nature of ganglion cell migration and maturation. One of them is the cephalo-caudal migration theory. [1] Some authors believe that the neuroblasts adjacent to pharynx arrive at esophagus by fifth week and at midgut by seventh week of gestation. The maturation of ganglion cells, however, continues up to second year of life. [1] Tam, in his study on human enteric innervations, [2] showed that the entire neural population as well as its substance P-ergic subpopulation followed a dual gradient of development, proceeding from each end to the middle of the gut. They found that at all ages studied, the foregut was most developed, the hind gut was less developed, and the ileum was least developed. According to him, the ganglion cells first appear in the myenteric plexus and then move inward. The ganglion cell migration has been studied at different levels and neural cell adhesion molecule (NCAM) is thought to play an important role in migration and localization of ganglion cells during embryogenesis. Kobayashi et al. [3] studied NCAM reactivity in Hirschsprung's Disease (HD) and confirmed it with age- and sex-matched controls and showed that intestines which contained ganglion cells had strong NCAM reactivity, whereas NCAM positive nerve fibers were absent in the myenteric plexus of aganglionic segments.

The aim of the study was 1) to examine the migration and developmental pattern of ganglion cells in fetuses aged 9-21 weeks; 2) to document whether the migration was occurring circumferentially equally in the entire axis or if there were discrepancies in different portions at the same level (i.e., same distance from anorectal junction), the data regarding which could be helpful in choosing a site of biopsy sample for diagnostic purpose and efficacy of surgical procedures.


   Materials and Methods Top


Fetuses from cases of spontaneous abortion or elective termination were taken. A total of 16 fetuses of gestational age from 9 to 21 weeks were studied. The whole gut was dissected out. Sections were taken from different parts of the gut including two from stomach, three from ileum and four from colon. Serial longitudinal sections from colon, the first one including the squamo-columnar junction, were studied. The sections were taken from four quadrants of the circumference of colonic lumen. Transverse sections were studied from small intestine and stomach. The sections were stained with Hematoxylin and Eosin (H and E). Immunohistochemistry was done using neurone-specific enolase (NSE) by Streptavidin Biotin method. The sections were studied for 1) the presence of ganglion cells in the segments, 2) nature of ganglia compared to ganglionic bowel of infants, and 3) migration pattern of ganglionic cells at different levels of intestine.


   Results Top


We studied 16 fetuses aged 9-21 weeks [Table 1]. We found poor NSE reactivity in 9-12 weeks. From 13 weeks, ganglion cells could be identified with NSE staining in the myenteric plexus. The ganglia, however, were small and primitive in nature. No submucosal staining was noted at this stage.
Table 1: Migration pattern and morphologic characteristics of fetal enteric nervous system

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In 14 th week fetuses, the ganglia were better developed taking NSE stain nicely. Some large ganglia could be identified [Figure 1]. Submucosal plexus could also be made out though the ganglia were not well developed.
Figure 1: Photomicrograph showing high-power view of a large ganglion in a 16 week fetus (NSE, ×400)

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At 15th week, the submucosal plexuses were better defined [Figure 2] and nerve trunks could be identified. Ganglion cells were seen within 1 mm from the squamo-columnar junction.
Figure 2: Photomicrograph depicting Auerbach's plexus of fetus showing collection of ganglion cells (18 weeks). (NSE, ×100)

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At 17 th week, the deep submucous plexuses were better defined and ganglion cells were identified 1.2 mm from squamo-columnar junction.

In the fetus of 19 weeks, well-formed ganglia were seen. Majority were mature ganglion cells. However, a few immature ganglion cells were visualized [Figure 3], better defined by NSE immunostaining. Ganglion cells could be identified within 1.5 mm of squamo-columnar junction.
Figure 3: Photomicrograph showing high-power view of fetal ganglia with immature ganglia cells (14 weeks). (H and E, ×400)

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At 21 weeks, both the plexuses were well developed [Figure 4]. But some immature ganglion cells remained. Ganglion cells could be identified within 2 mm of squamo-columnar junction.
Figure 4: Photomicrograph showing a ganglion in colonic submucosa in fetus (19 weeks). (NSE, ×400)

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Studying sections from different portions of gut wall including pylorus, ileum, and colon from the same fetus, we found that the first appearance of ganglion cells occurred in the myenteric plexus as early as 12 weeks in all of these sites [Graph 1]. [Additional file 1] However, well-developed ganglia appeared later in ileum (15 weeks) compared to pylorus (12 weeks) and colon (14 weeks) [Graph 2]. [Additional file 2]

First appearance of ganglion cells in deep submucosal plexus also coincided in all the three sites (i.e., pylorus, small intestine and colon). They were evident by 14 weeks of gestation. But well-developed ganglia were later to appear in the ileum (at 17 weeks) compared to pylorus (14 weeks) and colon (15 weeks). In superficial submucosal plexus also, ganglion cells appeared later (17 weeks) in the ileum compared to other sites (14 weeks at pylorus and 15 weeks at colon). Well-developed ganglia appeared as late as 18 weeks compared to 14 weeks and 15 weeks in pylorus and colon, respectively.

Overall, we noted that ganglion cells appear first in the myenteric plexus which is followed by deep and superficial submucous plexus. But when studied for the migration of ganglion cells around the circumference of gut wall, we found them to be at different levels from the squamo-columnar junction.


   Discussion Top


In our study with 16 fetuses of 9-21 weeks of gestation, we found that the ganglion cells even at the age of 12 weeks were present at the distal colonic segments but the distal aganglionic segment was not localized. Ganglion cells were found in the stomach, proximal small intestine and distal small intestine but the ganglia were not well defined. Well-defined ganglia with formation of interconnections were late to appear in the distal ileum. These findings support the dual migration hypothesis of enteric neurons. We cannot explain why there should be a single gradient in early differentiation of enteric neurons as shown by Okamoto and Ueda, [4] and a dual gradient of differentiation in older fetuses as shown in our study. Differences might be explained by differences in the period of intrauterine life studied and difference in methodology. In a study including similar intrauterine age fetuses as in our study, a dual migration gradient has been proposed. [5] A dual gradient of enteric neuronal development at various stages has similarly been observed in animal studies. These include the histochemical examinations of Cantino [6] in rats and rabbits, those of Keller [7] in chicks and more elegant experimental works of Le Douarin and Teillet [8] using the quail-chick chimeric system.

The formation of distal hypoganglionic segment was studied in detail taking serial longitudinal sections, the first one including the squamo-columnar junction. The length of distal hypoganglionic segment increased with increasing gestational age, which might be explained by the growth of the fetus. However, an interesting finding in our study was that the ganglion cells were not present at the same distance from the squamo-columnar junction in four quadrant longitudinal sections of a single case. This variation cannot be explained by dual migration hypothesis but this is an important finding to be kept in mind while taking biopsies for the diagnosis of HD or during the operative procedure.

NSE is a specific morphologic marker of all neurons with the distinctive property of being a functional indicator of differentiation. [9] NSE is an enzyme of known properties and has been fully characterized so that we know what exactly we are localizing. NSE is specific to neurons and absent in other elements of enteric nerve plexus. Appearance of NSE is also associated with definite developmental stage, i.e., cessation of cell division and development of synaptic contact. [2] Therefore, our findings with NSE localization of neurons support the dual migration theory and describe the nature of distal aganglionic segments in the fetuses of 9-21 weeks. Aganglionosis may occur as a result of defect in many stages of enteric neuronal development, including migration, colonization and maturation. Our finding of dual migration gradient calls for more inquests into the pathogenesis of HD. Hostile microenvironment theory [10] or immunological mechanisms may become the emerging theories of pathogenesis of HD.

In the present study, it was noted that development of enteric ganglia is delayed in ileum compared to rectum and stomach. This might be an indirect evidence in support of a dual migration pattern of intestinal ganglion cells. Studies analyzing the migration and maturation of fetal ganglia are limited in literature. The number of cases in our study is small. However, this finding might invite more studies on pathogenesis of HD. We also found that the level of distal aganglionic segments when measured from squamo-columnar junction varied with the age of gestation and the length was incongruous when studied at the same level from the squamo-columnar junction, taking four quadrant longitudinal sections. This finding might help surgeons while taking biopsies for diagnosis of HD or performing operative procedures in aganglionic segments.

 
   References Top

1.Smith B. Pre and post natal development of the rectum and its surgical implications. J Pediatr Surg 1968;3:386-94.  Back to cited text no. 1
[PUBMED]  [FULLTEXT]  
2.Tam PK. An immunohistochemical study with neurone-specific enolase and substance P of human enteric innervations: The normal developmental pattern and abnormal deviations in Hirschsprung's disease and pyloric stenosis. J Pediatr Surg 1986;21:227-32.  Back to cited text no. 2
[PUBMED]  [FULLTEXT]  
3.Kobayashi H, O'Brain BS, Puri P. Lack of expression of NADPH-diaphorase and neural cell adhesion molecule in colonic muscles of patients with Hirschsprung's disease. J Pediatr Surg 1994;29:301-4.  Back to cited text no. 3
    
4.Okamoto E, Ueda T. Embryogenesis of intramural ganglia of the gut and its relation to Hirschsprung's disease. J Pediatr Surg 967;2:437-43.  Back to cited text no. 4
    
5.Tam PK, Lister J. Developmental profile of neurone-specific enolase in human gut and its implications in Hirschsprung's disease. Gastroenterology 1986;90:1901-6.  Back to cited text no. 5
[PUBMED]    
6.Cantino D. A histological study of the nerve supply to the developing alimentary tract. Experientia 1970;15:766-7.   Back to cited text no. 6
    
7.Keller H. The development of the intramural nerve plexus of the gastrointestinal tract. Anat Embryol 1976;150:1-6.  Back to cited text no. 7
[PUBMED]    
8.Le Douarin NM, Teillet MA. Experimental analysis of the emigration and differentiation of the autonomic nervous system and of neuroectodermal mesenchymal derivatives, using a biological cell making technique. Dev Biol 1974;41:162-84.  Back to cited text no. 8
[PUBMED]  [FULLTEXT]  
9.Bishop AE, Polak JM, Facer P, Ferri GL, Marangos PJ, Pearse AG. Neuron specific enolase: A common marker for the endocrine cells and innervation of the gut and pancreas. Gastroenterology 1982;83:902-15.  Back to cited text no. 9
[PUBMED]    
10.Langer JC, Betti PA, Blennerhasset MG. Smooth muscle from aganglionic bowel in Hirschsprung's disease impairs neuronal development invitro. Cell Tissue Res 1994;276:181-6.  Back to cited text no. 10
    

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Correspondence Address:
Ranjana Bandyopadhyay
1B/3, Uttarpara Housing Estate, PO Bhadrakali, Dist. Hooghly, West Bengal - 712 232
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0377-4929.81589

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    Figures

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