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Pediatric Surgery International

, Volume 30, Issue 9, pp 901–906 | Cite as

Myogenin gene expression is not altered in the developing diaphragm of nitrofen-induced congenital diaphragmatic hernia

  • Toshiaki Takahashi
  • Florian Friedmacher
  • Hiromizu Takahashi
  • Alejandro Daniel Hofmann
  • Prem Puri
Original Article

Abstract

Purpose

Pleuroperitoneal folds (PPFs) represent the only source of muscle precursors cells (MPCs) in the primordial diaphragm. However, the exact pathogenesis of malformed PPFs and congenital diaphragmatic hernia (CDH) remains unclear. The muscle-specific transcription factor myogenin plays a key role during development and muscularization of the fetal diaphragm. Although myogenin knockout mice lack skeletal muscle fibers, the diaphragmatic musculature is intact without any defects. It has further been demonstrated that proliferation and differentiation of MPCs in PPFs and developing diaphragms are normal in rodent CDH models. We hypothesized that myogenin gene expression is not altered in malformed PPFs, developing diaphragms and diaphragmatic musculature in the nitrofen-induced CDH model.

Methods

Pregnant rats were exposed to nitrofen or vehicle on gestational day 9 (D9). Fetuses were harvested during PPF formation (D13), diaphragmatic development (D14-15) and muscularization (D18-21). Fetal PPFs, developing diaphragms and diaphragmatic musculature were dissected and divided into nitrofen and control groups. Myogenin mRNA levels were analyzed by quantitative real-time polymerase chain reaction, while immunohistochemistry was performed to investigate myogenin protein expression and distribution.

Results

Relative mRNA expression of myogenin was not significant different in PPFs (0.30 ± 0.09 vs. 0.48 ± 0.09; P = 0.37), developing diaphragms (1.25 ± 0.29 vs. 1.60 ± 0.32; P=0.53) and diaphragmatic musculature (1.08 ± 0.24 vs. 1.59 ± 0.20; P = 0.15) of nitrofen-exposed fetuses compared to controls. Myogenin immunoreactivity was not altered in the muscular components of malformed PPFs, developing diaphragms and diaphragmatic musculature of nitrofen-exposed fetuses compared to controls.

Conclusion

Myogenin gene expression is not altered in PPFs, developing diaphragms and diaphragmatic musculature in the nitrofen-induced CDH model, thus suggesting that diaphragmatic defects in this model develop independent of myogenic processes.

Keywords

Myogenin Pleuroperitoneal folds Diaphragm Congenital diaphragmatic hernia Nitrofen 

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Toshiaki Takahashi
    • 1
  • Florian Friedmacher
    • 1
  • Hiromizu Takahashi
    • 1
  • Alejandro Daniel Hofmann
    • 1
  • Prem Puri
    • 1
    • 2
  1. 1.National Children’s Research Centre, Our Lady’s Children’s HospitalDublin 12Ireland
  2. 2.Conway Institute of Biomolecular and Biomedical ResearchSchool of Medicine and Medical Science, University College DublinDublinIreland

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