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Pediatric Surgery International

, Volume 27, Issue 2, pp 159–163 | Cite as

Expression of the Wilm’s tumor gene WT1 during diaphragmatic development in the nitrofen model for congenital diaphragmatic hernia

  • Jens Dingemann
  • Takashi Doi
  • Elke Ruttenstock
  • Prem Puri
Original Article

Abstract

Purpose

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.

Conclusion

Downregulation of diaphragmatic WT1 gene expression may impair diaphragmatic development in the nitrofen CDH model.

Keywords

Congenital diaphragmatic hernia Nitrofen Pleuroperitoneal folds Diaphragmatic defect WT1 

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Jens Dingemann
    • 1
  • Takashi Doi
    • 1
  • Elke Ruttenstock
    • 1
  • Prem Puri
    • 1
  1. 1.National Children’s Research Centre, Our Lady’s Children’s Hospital DublinIreland University College DublinDublin 12Ireland

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