Abstract
PurposeThe aim of this study was to investigate the expression and significance of neuroligins in myenteric cells of Cajal (ICC-MY) in Hirschsprung’s disease (HSCR).MethodsLongitudinal muscle with adherent myenteric plexus (LMMP) from surgical excision waste colon of HSCR children were prepared by peeling off the mucous layer, sub-mucosal layer and circular muscle. Neuroligins, c-Kit (c-Kit-immunoreactivity representing ICC) and their relationship were assessed by double labeling immunofluorescence staining. ICC-MY were dissociated and cultured from LMMP by enzymolysis method, and were purified and analyzed using a combination of magnetic-activated cell sorting (MACS) and flow cytometry (FCM). Western-blot analysis was applied to compare and evaluate the expression levels of neuroligins in ICC-MY which were dissociated from different segments of HSCR (ganglionic colonic segment, transitional colonic segment and aganglionic colonic segment).ResultsNeuroligins and c-Kit were expressed on the same cells (ICC-MY); ICC-MY were dissociated, cultured and purified. For HSCR, neuroligins were expressed significantly in ICC-MY from ganglionic colonic segments, moderately in those from transitional colonic segments and down-regulated significantly in those from aganglionic colonic segments.ConclusionsNeuroligins were expressed in ICC-MY of human beings, and the expression varies from different segments of HSCR. This abnormal expression might play an important role in the pathogenesis of this disease through affecting the synaptic function of ICC-MY.
Highlights
Hirschsprung’s disease (HSCR) is a congenital condition that affects 1 of 5,000 human births and is characterized by colonic stasis due to the absence of enteric neurons in the distal gut [1], leading to tonic contraction of the affected segment, intestinal obstruction and massive distension of the proximal bowel
In order to explore these questions, we investigated the expression of neuroligins in interstitial cells of Cajal (ICC)-MY of children patients with HSCR by immunofluorescence and evaluated the expression levels of neuroligins in ICC-MY dissociated from different segments in HSCR by Western-blot analysis
Identification of ICC-MY in Longitudinal muscle with adherent myenteric plexus (LMMP) After immunohistochemical staining was performed on LMMP, light microscopy was used for observation
Summary
Hirschsprung’s disease (HSCR) is a congenital condition that affects 1 of 5,000 human births and is characterized by colonic stasis due to the absence of enteric neurons in the distal gut [1], leading to tonic contraction of the affected segment, intestinal obstruction and massive distension of the proximal bowel (magacolon). It has been identified that several genes such as RET, SOX10 and EDNRB are involved in the pathogenesis of HSCR in human beings [3]. ICC can be identified immunohistochemically by the expression of stem cell factor (SCF) receptor (c-Kit, CD117), a tyrosine kinase [7]. Designed cKit(CD117) antibodies have been developed to be the marker of ICC and ICC can be identified by their ability to bind to c-Kit(CD117) antibodies and by their clear morphological features[8,9]. ICC could be purified and analyzed by using a combination of magnetic-activated cell sorting (MACS) and flow cytometry (FCM) by the marker fraction as Kit+CD44+[10,11,12]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
