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


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Department of Pediatric Surgery, Hospital Universitario La Paz, Paseo de la Castellana, 261, 28046, Madrid, Spain. lmartinezm.hulp@salud.madrid.org
Leopoldo
L
Martínez
Rosa
R
Aras-López
Sara
S
Lancha
María Teresa
MT
Vallejo-Cremades
Federica
F
Pederiva
Liu
L
XiaoMei
Juan Antonio
JA
Tovar
D000818:Animals; D015153:Blotting, Western; D017615:Enteric Nervous System / Q000002:abnormalities* / Q000196:embryology / Q000201:enzymology; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006548:Hernia, Diaphragmatic / Q000196:embryology / Q000235:genetics; D065630:Hernias, Diaphragmatic, Congenital; D007150:Immunohistochemistry; D011247:Pregnancy; D011270:Pregnancy, Animal; D012313:RNA / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D043222:Ubiquitin Thiolesterase / Q000235:genetics / Q000378:metabolism
D012313:RNA; C489034:UCHL1 protein, rat; D043222:Ubiquitin Thiolesterase