Developmental changes in submucosal nitrergic neurons in the porcine distal colon

Our Ladys Childrens Hospital, Crumlin, Dublin, Leinster, Ireland
Journal of Pediatric Surgery (Impact Factor: 1.39). 05/2006; 41(5):1029-35. DOI: 10.1016/j.jpedsurg.2005.12.063
Source: PubMed


As our understanding of the enteric nervous system improves, it becomes clear that it is no longer sufficient to simply determine whether enteric ganglion cells are present but also to determine whether correct number and types of ganglion cells are present. Nitric oxide is recognized as a potent mediator of inhibitory nerves responsible for the relaxation of the smooth muscle of the gastrointestinal tract. The aim of this study was to determine the normal nitrergic neuronal density and morphology in the submucosal plexus of the porcine distal bowel from fetal life to adulthood.
Distal large bowel specimens were obtained from porcine fetuses of gestational age E60 (n = 5), E90 (n = 5), 1-day-old piglets (n = 5), 4-week-old piglets (n = 5), 12-week-old piglets (n = 5), and adult pigs (n = 5). Whole-mount preparations of the submucosal plexus were made and stained with NADPH diaphorase histochemistry. The ganglia density, the number of ganglion cells per ganglia, and nucleus and cytoplasmic area were measured.
Ganglia density decreased progressively and markedly with age until the adulthood (P < .001). On the contrary, ganglion cells increased their size over time predominantly because of increase in cytoplasm (P < .001). The number of ganglion cells per ganglia increased significantly during the fetal life. However, there was a significant reduction in the number of ganglion cells per ganglia during the period from birth to 4 weeks, remaining constant thereafter (P < .001).
The quantitative and qualitative morphometric analysis of the colonic submucous plexus shows that significant developmental changes occur during fetal and postnatal life. These findings indicate that the age of the patient is of utmost importance during histopathologic evaluation of enteric nervous system disorders.

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    • "The change of number of positive neurons per ganglia was dynamic, but the tendency was increased from 10-days-old to adult. These results are similar to the nitrergic neurons revealed by other researchers along the development (Montedonico et al., 2006; Sri Paran et al., 2008; Cserni et al., 2009b) in different species. It suggests that the relation of the major subpopulations of neurons is not changing significantly. "
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    ABSTRACT: The distribution of different subpopulations of enteric neurons in the developing enteric nervous system has been extensively examined in various animals, but up to date, there is no reference found regarding with the normal cholinergic neuronal density and morphology in the submucosal plexus of the developing chicken. In the present study, the fresh specimens were collected at day old, 5, 10, 20, and 40 days chickens respectively, for the whole-mount preparations of the submucosal plexus of chicken ileum. Acetylcholinesterase (AChE) histochemistry was performed. The morphology of the network, cytoplasmic area, the density of the ganglia, and number of the ganglion cells per ganglia was measured. Our results reported that the positively stained neurons have various morphologies and staining intensity. They formed a clear network by ganglia and nerve fibers. The number of ganglia per unit area decreased with age, however the size of positive neurons increased, as the size, in 40 day old group was 3 times than that of the day old group. Change the number of positive neurons per ganglia was dynamic, but the tendency was increased from 10-days old to adult chickens. It was concluded that the significant developmental changes occur in postnatal life of chicken ileal submucosal plexus. The changes of the cholinergic neuronal subpopulation are similar than it has been found in the nitrergic subpopulation in different animal models.
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    • "Thus, we tried to provide a better study on ENS connecting conventional sections and whole-mount preparations. Junquera et al, (1998) reported that the AChE and NOS positive neurons varied from irregular or polygonal in shape in rabbit intestine, in addition, the size of AChE and NOS positive neurons were different with other animals (An et al., 2003; Montedonico et al., 2006; Cserni et al., 2007; Liu et al., 2007). Some studies revealed small intestine has a well-developed plexus and ganglia are aligned along the longitudinal axis of the gut and has a highest density in ileum (Liu et al., 2007). "
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    ABSTRACT: Histochemical loacalization and analysis of the cholinergic and nitrergic neurons in the chicken ileum were investigated by staining with acetylcholine esterases (AChE) and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d), respectively. AChE and NADPH-d activity was demonstrated in neuronal cell bodies and nerve fibres in the chicken ileum. The positive neurons showed irregular or polygonal shape and were mainly present sporadic or clumped in the myenteric and submucosal plexus. The positive nerve fibres frequently surrounded the ileac blood vessels. They were abundantly present in myenteric and submucosal plexus of the ileum forming a network. Some positive nerve fibres traversed the submucosa into the lamina propria mucosae. Fine nerve fibres were found to penetrate into intestinal villi underneath the epithelium. Extensive networks of more intensely staining AChE positive nerve fibers were present in the mucosa as compared to that of NADPH-d positive fibers. Ganglia density of submucosal plexus was markedly bigger than that of myenteric plexus, whereas neurons per ganglion and the number of neurons per mm2 and the size of neurons of submucosal plexus were shorter than that of myenteric plexus. In addition, the number of AChE positive neurons and nerve fibres was more than that of NADPH-d positive neurons and nerve fibres. We concluded that the chicken ileum is characterized by abundance of nerve structures which may play a significant functional role in ileum of the chicken.
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