L. Nemeth et P. Puri, The innervation of human bowel mucosa and its alterations in Hirschsprung's disease using a whole-mount preparation technique, PEDIAT SURG, 16(4), 2000, pp. 277-281
The innervation of the human bowel wall and its structural and functional c
hanges in Hirschsprung's disease (HD) are well-recognised, The luminal surf
ace of the bowel acts as a multifunctional barrier, and modifications in it
s physiochemical properties can result in serious complications such as ent
erocolitis (EC). The whole-mount preparation (WMP) technique produces a thr
ee-dimensional (3D) picture to better demonstrate the neuronal networks and
the relationship of branching and interconnecting nerve fibres to each oth
er. The aim of this study was to investigate the innervation of the mucosal
layer in normal and HD bowel using a WMP immunohistochemistry technique in
order to better understand the pathophysiology of WD. Full-thickness bowel
specimens were collected from 9 HD patients at pull-through operation. Nor
mal control small- and large-bowel specimens were collected from 10 patient
s at the time of bladder augmentation. Suction rectal biopsies from 8 patie
nts with chronic constipation and 2 patients with HD were also included in
this study. A WMP of the mucosal layer was made and stained with various ne
uronal markers (S100, PCP 9.5, and LlCAM) using fluorescein immunohistochem
istry. PGP 9.5, S100, and LlCAM immunofluorescence staining of the normal m
ucosa demonstrated a characteristic 3D meshlike neuronal network of uniform
thickness surrounding the crypts. In the aganglionic bowel S100, PGP 9.5,
and LlCAM-positive meshlike networks were replaced by thick nerve trunks in
the muscosa without any interconnecting network. The present study demonst
rates for the first time the 3D morphology of mucosal innervation in normal
and aganglionic bowel. The WMP technique clearly demonstrated that the muc
osal innervation in HD is morphologically abnormal, and this may adversely
influence secretory and absorptive functions of the bowel. WMPs using sucti
on rectal biopsy specimens may be a useful additional technique to diagnose
HD.