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Pediatric Surgery International

, Volume 27, Issue 6, pp 579–582 | Cite as

The role of primary myogenic regulatory factors in the developing diaphragmatic muscle in the nitrofen-induced diaphragmatic hernia

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

Abstract

Purpose

The nitrofen model of congenital diaphragmatic hernia (CDH) is widely used to investigate the pathogenesis of CDH. However, the exact pathomechanism of the diaphragmatic defect is still unclear. Diaphragmatic muscularization represents the last stage of diaphragmatic development. Myogenic differentiation 1 (MyoD) and myogenic factor 5 (Myf5) play a crucial role in muscularization. MyoD−/− : Myf5+/− mutant mice show reduced diaphragmatic size, whereas MyoD+/− : Myf5−/− mutants have normal diaphragms. We designed this study to investigate diaphragmatic gene expression of MyoD and Myf5 in the nitrofen CDH model.

Methods

Pregnant rats received nitrofen or vehicle on day 9 of gestation (D9), followed by cesarean section on D18 and D21. Fetal diaphragms (n = 40) were micro-dissected and divided into CDH group and controls. MyoD and Myf5 mRNA-expression were determined using Real-time PCR. Immunohistochemistry was performed to evaluate protein expression of MyoD and Myf5.

Results

Relative diaphragmatic mRNA expression levels and immnunoreactivity of MyoD were decreased in the CDH group on D18 and D21. Myf 5 mRNA and protein expression were not altered in the CDH group.

Conclusion

This is the first study showing that MyoD expression is selectively decreased in the diaphragm muscle in the nitrofen model of CDH.

Keywords

Congenital diaphragmatic hernia Nitrofen Diaphragmatic defect, MyoD, Myf5 

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Jens Dingemann
    • 1
  • Takashi Doi
    • 1
    • 2
  • Elke Ruttenstock
    • 1
  • Prem Puri
    • 1
    • 2
  1. 1.National Children’s Research CentreOur Lady’s Children’s HospitalDublin 12Ireland
  2. 2.UCD School of Medicine and Medical SciencesUniversity College DublinDublinIreland

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