Cholinergic innervation and ganglion cell distribution in Hirschsprung\u2019s disease

Anne K Braczynski,Michel Mittelbronn,Udo Rolle,Stefan Gfroerer,Peter Baumgarten,Patrick N Harter,Rudi Beschorner

doi:10.1186/s12887-020-02299-z

Abstract

BackgroundThe diagnostic gold standard of Hirschsprung’s disease (HD) is based on the histopathological assessment of colorectal biopsies. Although data on cholinergic innervation and ganglion cell (GC) distribution exist, only few studies have examined these two key features together. We assessed the pattern of cholinergic innervation and the amount of GCs in colorectal specimens of 14 HD patients.MethodsWe established a semi-quantitative score for cholinergic innervation using acetylcholinesterase (AChE) enzyme histochemistry and quantitatively analyzed the number of GCs via NADH tetrazolium reductase (NADH) enzyme histochemistry. We examined both the entire length of the resected specimens as well as defined areas of the transition zone of both pathological and healthy appearing segment.ResultsHigh AChE score values were associated with absence of GCs, and AChE scores were inversely correlated with the number of GCs. Nevertheless, we observed several cases in which one of the two features revealed a normal distribution pattern, whereas the other still displayed pathological features.ConclusionsOur data support the need for transmural colon biopsies, to enable the best evaluation of both cholinergic innervation and GCs for a reliable assessment of HD.

Highlights

The diagnostic gold standard of Hirschsprung’s disease (HD) is based on the histopathological assessment of colorectal biopsies
AChE score negatively correlated with ganglion cell count In areas with no AChE-positive nerve fibers in the mucosal layer, a median of 17.8 GCs/ mm was observed
Low AChE score values were associated with a high number of ganglion cells, but the absence of pathological cholinergic innervation was not a reliable indicator for a regular ganglion cell count Moderate or severe pathological cholinergic innervation in the Lamina propria mucosae (LP) was associated with complete absence of ganglion cells in the myenteric plexus (Fig. 4b and Fig. 5g-i)

Summary

Introduction

The diagnostic gold standard of Hirschsprung’s disease (HD) is based on the histopathological assessment of colorectal biopsies. Data on cholinergic innervation and ganglion cell (GC) distribution exist, only few studies have examined these two key features together. We assessed the pattern of cholinergic innervation and the amount of GCs in colorectal specimens of 14 HD patients. Hirschsprung’s disease (HD) is a rare cause of constipation in infants that is microscopically characterized by congenital absence of GCs in a variable extension mainly affecting the rectum and sigmoid colon. A register-based study has described a slight increase in incidence over the recent years [2]. 50 different RET mutations have been reported in HD patients, accounting for 50% of familial cases of HD and 15 to 20% of sporadic cases of HD [7]

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Conclusion