Pediatric Surgery International

, Volume 35, Issue 2, pp 209–214 | Cite as

Innervation of the entire internal anal sphincter in a mouse model of Hirschsprung’s disease: a first report

  • Masahiro Takeda
  • Katsumi MiyaharaEmail author
  • Ryo Sueyoshi
  • Atsushi Arakawa
  • Geoffrey J. Lane
  • Atsuyuki Yamataka
Original Article



Impaired function of the internal anal sphincter (IAS) may be implicated in postoperative obstructed defecation (POD) that may complicate Hirschsprung’s disease (HD) patients. While innervation of part of the IAS in HD has been reported, accurate details based on anatomic landmarks that can explain the clinical morbidity seen in POD are lacking, and there appear to be no studies that specifically document the innervation of the “entire” IAS in HD. We used endothelin receptor-B knockout mice to represent HD (HD-mice) and C57B6 wild mice as controls (C-mice) to investigate the innervation of the entire IAS to assess the pathophysiology of POD experimentally.


The end-point of the longitudinal muscle layer was used to define the border between the IAS and the circular muscle layer (CML). Specimens of anorectum from HD- and C-mice were immunostained with PGP 9.5 and S100 as general nerve markers, nNOS and VIP as parasympathetic nerve markers, TH as a sympathetic nerve marker, and calretinin as a reliable diagnostic marker for HD. Immunostained cells/fibers were quantified using ImageJ.


On fluorescence microscopy, PGP 9.5, nNOS, and calretinin were significantly lower in the IAS of HD-mice than in C-mice (p < 0.05, respectively), while there were no significant differences between HD-mice and C-mice for S100, VIP, or TH.


We are the first to confirm that the expression of histochemical markers of innervation is abnormal throughout the “entire” IAS in HD-mice. Application of this finding may be beneficial for preventing POD and requires further research.


Hirschsprung’s disease Internal anal sphincter Endothelin receptor-B knockout mice 



This work was supported by JSPS KAKENHI Grant Numbers 15K10929 and 17K17004.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Masahiro Takeda
    • 1
  • Katsumi Miyahara
    • 1
    Email author
  • Ryo Sueyoshi
    • 1
  • Atsushi Arakawa
    • 2
  • Geoffrey J. Lane
    • 1
  • Atsuyuki Yamataka
    • 1
  1. 1.Department of Pediatric General and Urogenital SurgeryJuntendo University School of MedicineTokyoJapan
  2. 2.Department of Human PathologyJuntendo University School of MedicineTokyoJapan

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