Acetylcholinesterase histochemistry (AChE) - A helpful technique in the diagnosis and in aiding the operative procedures of Hirschsprung disease

Abstract

BackgroundHirschsprung’s disease (HD) is an anomaly characterized by the absence of myenteric and submucosal ganglion cells (GC) in the distal alimentary tract. Diagnosis of HD is made by the absence of GC and missing out on even a single ganglion cell can be very devastating. Acetylcholinesterase (AChE) histochemistry, done on frozen sections is said to be a very useful ancillary technique in the diagnosis and in aiding the operative procedures of HD.MethodsTo assess this, 73 samples from 42 suspected/known cases of HD were subjected to frozen section analysis with rapid haematoxylin and eosin, toluidin blue stain along with AChE histochemistry. The remnant sample was paraffin embedded for routine haematoxylin and eosin staining.ResultsOn frozen section analysis, 33 samples showed absence of ganglion cells, AChE histochemistry showed a positive staining pattern in 17 samples and paraffin embedded routine, H&E stained sections showed absence of ganglion cells in 19 samples. Sensitivity and specificity of both tests ie frozen section rapid H&E/AChE histochemistry in the diagnosis of HD, were calculated taking paraffin embedded H&E stained sections as the gold standard. Sensitivity of frozen section rapid H&E in the diagnosis of HD is 57.57 % and specificity is 79.10 %. The p-value is <0.0001, which is significant. The sensitivity of AChE histochemistry in the diagnosis of HD is 90.47 % and specificity is 96.36 %. The p-value is <0.0001, which is significant.ConclusionsAcetylcholineesterase (AChE) histochemistry is a very useful ancillary technique in the diagnosis and in aiding the operative procedures of HD. It acts as a double check in the diagnosis of HD.

The role of the α7nAChR-mediated cholinergic anti-inflammatory pathway in Hirschsprung associated enterocolitis

“…The main receptor for acetylcholine on immune cells is α7nAChR, which is expressed in a variety of immune cells [17,18,19]. Studies [20]also found that the distributions of intestinal cholinergic nerve fibers were tightly associated with macrophages with positive expression of α7nAChR, which laid a morphological foundation for acetylcholine to inhibit the activation of macrophages through α7nAChR.…”

Transanal endorectal stepwise gradient muscular cuff cutting pull‑through method: Technique refinements and comparison with laparoscopy‑assisted procedures

“…In the first step, multiple seromuscular leveling biopsies of the bowel were obtained in order to pathologically detect ganglion cells in the submucosal nerve plexus. The length of abscission was also determined based on biopsies that confirmed the absence of ganglion cells in sections of the colon; typical histological findings for HD in a rectal biopsy include the absence of ganglion cells in the submucosal (Meissner's) plexus and the intermuscular (Auerbach's) plexus (17,18). The mesentery was separated from the rectal wall to protect the lower rectal vessels and maintain a good blood supply to the rectal muscular sheath.…”

Is hematoxylin-eosin staining in rectal mucosal and submucosal biopsies still useful for the diagnosis of Hirschsprung disease?

“…This fragment may be obtained less invasively compared to the tissue sampling needed for HE staining. This histochemical technique uses the fact that children with HD have increased AChE activity in their rectal mucosal and submucosal biopsy fragments [ 7 – 9 ]. This method is considered the gold standard for the diagnosis of HD in our practice, and it exhibits an accuracy greater than 90%.…”