title: Expression of the Wilm's tumor gene WT1 during diaphragmatic development in the nitrofen model for congenital diaphragmatic hernia.
abstract: OBJECTIVE
  The nitrofen model of congenital diaphragmatic hernia (CDH) reproduces a typical diaphragmatic defect. However, the exact pathomechanism of CDH is still unknown. The Wilm's tumor 1 gene (WT1) is crucial for diaphragmatic development. Mutations in WT1 associated with CDH have been described in humans. Additionally, WT1(-/-) mice display CDH. Furthermore, WT1 is involved in the retinoid signaling pathway, a candidate pathway for CDH. We hypothesized that diaphragmatic WT1 gene expression is downregulated during diaphragmatic development in the nitrofen CDH model.
  
  
  METHODS
  Pregnant rats received vehicle or nitrofen on gestational day 9 (D9). Embryos were delivered on D13, D18 and D21. The pleuroperitoneal folds (PPFs) were dissected using laser capture microdissection (D13). Diaphragms of D18 and D21 were manually dissected. RNA was extracted and relative mRNA expression of WT1 was determined using real-time PCR. Immunofluorescence was performed to evaluate protein expression of WT1. Statistical significance was considered p < 0.05.
  
  
  RESULTS
  Diaphragmatic mRNA expression of WT1 was significantly decreased in the nitrofen group on D13, D18 and D21. Intensity of immunofluorescencence of WT1 was markedly decreased in the CDH diaphragms on D13, D18 and D21.
  
  
  CONCLUSIONS
  Downregulation of diaphragmatic WT1 gene expression may impair diaphragmatic development in the nitrofen CDH model.
authors.affiliation: National Children's Research Centre, Our Lady's Children's Hospital Dublin, Ireland University College Dublin, Dublin 12, Ireland.
authors.forename: Jens
authors.initials: J
authors.lastname: Dingemann
authors.forename: Takashi
authors.initials: T
authors.lastname: Doi
authors.forename: Elke
authors.initials: E
authors.lastname: Ruttenstock
authors.forename: Prem
authors.initials: P
authors.lastname: Puri
mesh_terms: D000818:Animals; D003964:Diaphragm / Q000187:drug effects / Q000196:embryology* / Q000378:metabolism; D004195:Disease Models, Animal; D005260:Female; D018507:Gene Expression Regulation, Developmental; D016162:Genes, Wilms Tumor; D006548:Hernia, Diaphragmatic / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism; D065630:Hernias, Diaphragmatic, Congenital; D010647:Phenyl Ethers / Q000633:toxicity; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011270:Pregnancy, Animal; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D010647:Phenyl Ethers; D012333:RNA, Messenger; C007350:nitrofen