Pediatric Surgery International

, Volume 27, Issue 2, pp 165–173 | Cite as

Abnormal development of the enteric nervous system in rat embryos and fetuses with congenital diaphragmatic hernia

  • Leopoldo MartínezEmail author
  • Rosa Aras-López
  • Sara Lancha
  • María Teresa Vallejo-Cremades
  • Federica Pederiva
  • Liu XiaoMei
  • Juan Antonio Tovar
Original Article



Esophageal dilatation, gastroesophageal reflux, and intestinal obstruction have been demonstrated in CDH survivors. Abnormal esophageal and intestinal innervations were recently found in rats and babies with this disease. Our aim was to further characterize these malformations in embryos and fetal rats exposed to nitrofen.


Pregnant rats received either 100 mg nitrofen or vehicle on E9.5. Fetuses were recovered at E15, E18, and E21. Sections of esophagus and small bowel were histochemically stained with acetylcholinesterase (AChE) and immunostained for PGP9.5. PGP9.5 gen protein were measured on E21 and PGP9.5 mRNA on E15, E18 and E21. Comparisons between groups were made with non-parametrics tests.


Histochemistry and immunohistochemistry showed deficient innervation in all anatomical areas studied at E15, E18, and E21, and WB confirmed this decrease in E21 fetuses. PGP9.5 messenger was decreased in nitrofen-exposed animals on E18 (esophagus) or E15 (small bowel), and increased on E21 in the esophagus and E18 in small bowel.


Development of the enteric nervous system of the esophagus, stomach, and small bowel is deficient in rat embryos and fetuses exposed to nitrofen. These anomalies could account in part for the long-term gastrointestinal morbidity observed in CDH survivors.


Congenital diaphragmatic hernia Nitrofen Enteric nervous system PGP9.5 Histochemistry Acetylcholinesterase Immunohistochemistry Western blot RT-PCR 



Supported in part by FIS (06/0486 and 06/0447), IdiPaz and FMM Grants and by the Spanish Health Institute Carlos III (grant No. RD08/0072: Maternal, Child Health and Development Network).


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Leopoldo Martínez
    • 1
    Email author
  • Rosa Aras-López
    • 2
  • Sara Lancha
    • 2
  • María Teresa Vallejo-Cremades
    • 2
  • Federica Pederiva
    • 1
  • Liu XiaoMei
    • 3
    • 4
  • Juan Antonio Tovar
    • 1
  1. 1.Department of Pediatric SurgeryHospital Universitario La PazMadridSpain
  2. 2.Research Laboratory IdipazHospital Universitario La PazMadridSpain
  3. 3.Central LaboratoryShengjing Hospital of China Medical UniversityShenyangChina
  4. 4.Research Laboratory IdiPazHospital Universitario La PazMadridSpain

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