Related Experiment Video
Updated: May 28, 2026
Immunostaining to Visualize Murine Enteric Nervous System Development
Published on: April 29, 2015
Abnormal development of intrinsic innervation in murine embryos with anorectal malformations
Yoshihiro Kubota1, Hirotomi Cho, Tomoko Umeda
1Department of Surgery, Shiga University of Medical Science, Setatsukinowa-cho, Otsu, Shiga, Japan. kubyos@belle.shiga-med.ac.jp
Insights
Reduced expression of substance P (SP), vasoactive intestinal peptide (VIP), and c-kit in the rectal muscle layer of mouse embryos with anorectal malformations (ARMs) may lead to severe constipation after surgery.
Area of Science:
- Developmental biology
- Gastroenterology
- Pediatric surgery
Background:
- Anorectal malformations (ARMs) often lead to constipation, soiling, and incontinence post-surgical repair.
- Investigating the molecular mechanisms underlying these functional deficits is crucial for improving patient outcomes.
Purpose of the Study:
- To examine the expression of substance P (SP), vasoactive intestinal peptide (VIP), and c-kit in the developing rectum of mouse embryos with induced ARMs.
Main Methods:
- Anorectal malformations were induced in mouse embryos using etretinate on embryonic day 9.
- Immunohistochemical staining was used to assess the expression of SP, VIP, and c-kit in rectal tissues between embryonic days 14 and 18.
Main Results:
- Decreased SP and VIP immunoreactivity were observed in the rectal circular muscle layer from embryonic day 16 in etretinate-treated embryos compared to controls.
- C-kit immunoreactivity was significantly lower in the rectal circular muscle layer of etretinate-treated embryos by embryonic day 18.
Conclusions:
- Reduced expression of SP, VIP, and c-kit in the rectal circular muscle layer of embryos with ARMs may contribute to severe constipation after surgical correction.
- These findings suggest potential therapeutic targets for managing functional bowel problems in children with ARMs.
Introduction:
Constipation, soiling, and incontinence are common problems after definitive repair of anorectal malformations (ARMs) in children. We studied the expression of substance P (SP), vasoactive intestinal peptide (VIP), and c-kit in the rectum of murine embryos with or without ARMs at later developmental stages.
Methods:
On the 9th embryonic day (E9), pregnant Institute of Cancer Research mice were fed etretinate, a synthetic vitamin A analogue (60 mg/kg), whereas controls were fed only with sesame oil. Embryos were excised between E14 and E18, and prepared for histological examination. The SP, VIP, and c-kit expressions were examined by immunohistochemical staining for the SP, VIP, and c-kit antigens, respectively.
Results:
On E14 and E15, the expression levels of the anti-SP and anti-VIP antibodies in the rectum did not differ between the control and etretinate-treated group. However, as compared to the controls, a decreased SP and VIP immunoreactivity was observed in the circular muscle layer of the rectum between E16 and E18. On the other hand, on E14 and E15, the expression of anti-c-kit antibody in the rectum did not differ between the etretinate-treated and control group. However, c-kit immunoreactivity was slightly higher in the circular muscle layer of the rectum in the controls on E16 and E17, and considerably higher on E18 than that of the muscle layer in the etretinate-treated group.
Conclusion:
At later developmental stages, the expression levels of SP, VIP, and c-kit reduced in the circular muscle layer of the rectum in mice with etretinate-induced ARMs. This result indicates that reduced SP, VIP, and c-kit expression levels in the circular muscle layer may cause severe constipation in children who develop severe ARMs after definitive surgery.
More Related Videos
Related Concept Videos
01:30Neurulation
01:15Development of the Sexual Organs in the Embryo and Fetus
Near the gonadal ridges, two duct systems are present: the mesonephric ducts (Wolffian ducts) and paramesonephric ducts (Müllerian ducts). These ducts form the basis for the male...