Regenerative Capacity of the Enteric Nervous System after Ileoileal Anastomoses in a Rat Model

Abstract

Purpose The aim of the study was to investigate the regeneration and migration of neuronal progenitor cells of the enteric nervous system during wound healing after intestinal anastomosis in the rat ileum.

Methods Experiments were performed in a rat model of ileoileal anastomosis. Rats were humanely killed on day 2 or day 10 after anastomosis, and the anastomotic region was compared with ileum of healthy rats. Immunofluorescent staining was performed with protein gene product 9.5, nestin, and S100 antibodies. Ganglia of the anastomotic region in both the myenteric and submucosal plexus were counted, and their diameters were measured and compared between groups.

Results Analysis of number and diameter of ganglia in both myenteric and submucosal plexus showed individual alterations as a reaction to the surgical manipulation. Significantly less ganglia were found in the submucosal plexus in the operated groups at both day 2 (p < 0.01) and day 10 (p < 0.01) than in the control group. In the myenteric plexus in the operated group, there was a difference in the number of ganglia at day 2, but ganglia count had recovered at day 10 and was not significantly different from the control group. However, the diameter of ganglia in the myenteric plexus still significantly decreasing on day 10 after surgery than in the control group (p = 0.046). Nestin and S100 double-staining showed an increased expression of nestin around the anastomotic wound.

Conclusion Our findings suggest a regenerative potential of the enteric nervous system after the surgical ileoileal anastomosis. The myenteric plexus appears to recover faster than the submucosal plexus. This recovery might be driven by nestin-positive neuronal progenitor cells.

Keywords

enteric nervous system - nestin - neuronal regeneration - intestinal anastomosis

• References

• 1 Furness JB. The enteric nervous system: normal functions and enteric neuropathies. Neurogastroenterol Motil 2008; 20 (Suppl. 01) 32-38
• 2 Furness JB. The Enteric Nervous System. Malden, MA: Blackwell; 2006
• 3 Wallace AS, Burns AJ. Development of the enteric nervous system, smooth muscle and interstitial cells of Cajal in the human gastrointestinal tract. Cell Tissue Res 2005; 319 (3) 367-382
• 4 Sanovic S, Lamb DP, Blennerhassett MG. Damage to the enteric nervous system in experimental colitis. Am J Pathol 1999; 155 (4) 1051-1057
• 5 Shen KZ, Surprenant A. Somatostatin-mediated inhibitory postsynaptic potential in sympathetically denervated guinea-pig submucosal neurones. J Physiol 1993; 470: 619-635
• 6 Hanani M, Ledder O, Yutkin V , et al. Regeneration of myenteric plexus in the mouse colon after experimental denervation with benzalkonium chloride. J Comp Neurol 2003; 462 (3) 315-327
• 7 Schäfer KH, Van Ginneken C, Copray S. Plasticity and neural stem cells in the enteric nervous system. Anat Rec (Hoboken) 2009; 292 (12) 1940-1952
• 8 Joseph NM, He S, Quintana E, Kim YG, Núñez G, Morrison SJ. Enteric glia are multipotent in culture but primarily form glia in the adult rodent gut. J Clin Invest 2011; 121 (9) 3398-3411
• 9 Laranjeira C, Sandgren K, Kessaris N , et al. Glial cells in the mouse enteric nervous system can undergo neurogenesis in response to injury. J Clin Invest 2011; 121 (9) 3412-3424
• 10 Wiese C, Rolletschek A, Kania G , et al. Nestin expression—a property of multi-lineage progenitor cells?. Cell Mol Life Sci 2004; 61 (19–20): 2510-2522
• 11 Vanderwinden JM, Gillard K, De Laet MH, Messam CA, Schiffmann SN. Distribution of the intermediate filament nestin in the muscularis propria of the human gastrointestinal tract. Cell Tissue Res 2002; 309 (2) 261-268
• 12 Tokui K, Sakanaka M, Kimura S. Progressive reorganization of the myenteric plexus during one year following reanastomosis of the ileum of the guinea pig. Cell Tissue Res 1994; 277 (2) 259-272
• 13 Yanagida H, Yanase H, Sanders KM, Ward SM. Intestinal surgical resection disrupts electrical rhythmicity, neural responses, and interstitial cell networks. Gastroenterology 2004; 127 (6) 1748-1759
• 14 Jabaji Z, Stark R, Dunn JC. Regeneration of enteric ganglia in mechanically lengthened jejunum after restoration into intestinal continuity. J Pediatr Surg 2013; 48 (1) 118-123
• 15 Mei F, Yu B, Ma H, Zhang HJ, Zhou DS. Interstitial cells of Cajal could regenerate and restore their normal distribution after disrupted by intestinal transection and anastomosis in the adult guinea pigs. Virchows Arch 2006; 449 (3) 348-357
• 16 Kruger GM, Mosher JT, Bixby S, Joseph N, Iwashita T, Morrison SJ. Neural crest stem cells persist in the adult gut but undergo changes in self-renewal, neuronal subtype potential, and factor responsiveness. Neuron 2002; 35 (4) 657-669
• 17 Gershon MD, Epstein ML, Hegstrand L. Colonization of the chick gut by progenitors of enteric serotonergic neurons: distribution, differentiation, and maturation within the gut. Dev Biol 1980; 77 (1) 41-51