Skip to main content
Top
Published in: Pediatric Surgery International 6/2011

01-06-2011 | Original Article

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

Authors: Jens Dingemann, Takashi Doi, Elke Ruttenstock, Prem Puri

Published in: Pediatric Surgery International | Issue 6/2011

Login to get access

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.
Literature
1.
go back to reference Gaxiola A, Varon J, Valladolid G (2009) Congenital diaphragmatic hernia: an overview of the etiology and current management. Acta Paediatr 98:621–627PubMedCrossRef Gaxiola A, Varon J, Valladolid G (2009) Congenital diaphragmatic hernia: an overview of the etiology and current management. Acta Paediatr 98:621–627PubMedCrossRef
3.
go back to reference Stege G, Fenton A, Jaffray B (2003) Nihilism in the 1990s: the true mortality of congenital diaphragmatic hernia. Pediatrics 112:532–535PubMedCrossRef Stege G, Fenton A, Jaffray B (2003) Nihilism in the 1990s: the true mortality of congenital diaphragmatic hernia. Pediatrics 112:532–535PubMedCrossRef
4.
go back to reference Clugston RD, Greer JJ (2007) Diaphragm development and congenital diaphragmatic hernia. Semin Pediatr Surg 16:94–100PubMedCrossRef Clugston RD, Greer JJ (2007) Diaphragm development and congenital diaphragmatic hernia. Semin Pediatr Surg 16:94–100PubMedCrossRef
5.
go back to reference Mortell A, Montedonico S, Puri P (2006) Animal models in pediatric surgery. Pediatr Surg Int 22:111–128PubMedCrossRef Mortell A, Montedonico S, Puri P (2006) Animal models in pediatric surgery. Pediatr Surg Int 22:111–128PubMedCrossRef
6.
go back to reference Clugston RD, Zhang W, Greer JJ (2008) Gene expression in the developing diaphragm: significance for congenital diaphragmatic hernia. Am J Physiol Lung Cell Mol Physiol 294:665–675CrossRef Clugston RD, Zhang W, Greer JJ (2008) Gene expression in the developing diaphragm: significance for congenital diaphragmatic hernia. Am J Physiol Lung Cell Mol Physiol 294:665–675CrossRef
7.
go back to reference Dingemann J, Doi T, Ruttenstock E, Puri P (2010) Downregulation of Fgfrl1 contributes to the development of the diaphragmatic defect in the nitrofen model for congenital diaphragmatic hernia. Eur J Pediatr Surg (in press) Dingemann J, Doi T, Ruttenstock E, Puri P (2010) Downregulation of Fgfrl1 contributes to the development of the diaphragmatic defect in the nitrofen model for congenital diaphragmatic hernia. Eur J Pediatr Surg (in press)
8.
go back to reference Dingemann J, Doi T, Ruttenstock E, Puri P (2010) Expression of the Wilm’s tumor gene WT1 during diaphragmatic development in the nitrofen model for congenital diaphragmatic hernia. Pediatr Surg Int (in press) Dingemann J, Doi T, Ruttenstock E, Puri P (2010) Expression of the Wilm’s tumor gene WT1 during diaphragmatic development in the nitrofen model for congenital diaphragmatic hernia. Pediatr Surg Int (in press)
9.
go back to reference Inanlou MR, Dhillon GS, Belliveau AC, Reid GA, Ying C, Rudnicki MA, Kablar B (2003) A significant reduction of the diaphragm in mdx:MyoD−/− (9th) embryos suggests a role for MyoD in the diaphragm development. Dev Biol 261:324–336PubMedCrossRef Inanlou MR, Dhillon GS, Belliveau AC, Reid GA, Ying C, Rudnicki MA, Kablar B (2003) A significant reduction of the diaphragm in mdx:MyoD−/− (9th) embryos suggests a role for MyoD in the diaphragm development. Dev Biol 261:324–336PubMedCrossRef
10.
go back to reference Kablar B, Rudnicki MA (2000) Skeletal muscle development in the mouse embryo. Histol Histopathol 15:649–656PubMed Kablar B, Rudnicki MA (2000) Skeletal muscle development in the mouse embryo. Histol Histopathol 15:649–656PubMed
11.
go back to reference Inanlou MR, Kablar B (2003) Abnormal development of the diaphragm in mdx:MyoD−/− (9th) embryos leads to pulmonary hypoplasia. Int J Dev Biol 47:363–371PubMed Inanlou MR, Kablar B (2003) Abnormal development of the diaphragm in mdx:MyoD−/− (9th) embryos leads to pulmonary hypoplasia. Int J Dev Biol 47:363–371PubMed
12.
go back to reference Holder AM, Klaassens M, Tibboel D, de Klein A, Lee B, Scott DA (2007) Genetic factors in congenital diaphragmatic hernia. Am J Hum Genet 80:825–845PubMedCrossRef Holder AM, Klaassens M, Tibboel D, de Klein A, Lee B, Scott DA (2007) Genetic factors in congenital diaphragmatic hernia. Am J Hum Genet 80:825–845PubMedCrossRef
13.
go back to reference Rudnicki MA, Schnegelsberg PN, Stead RH, Braun T, Arnold HH, Jaenisch R (1993) MyoD or Myf-5 is required for the formation of skeletal muscle. Cell 75:1351–1359PubMedCrossRef Rudnicki MA, Schnegelsberg PN, Stead RH, Braun T, Arnold HH, Jaenisch R (1993) MyoD or Myf-5 is required for the formation of skeletal muscle. Cell 75:1351–1359PubMedCrossRef
14.
go back to reference Babiuk RP, Greer JJ (2002) Diaphragm defects occur in a CDH hernia model independently of myogenesis and lung formation. Am J Physiol Lung Cell Mol Physiol 283:L1310–L1314PubMed Babiuk RP, Greer JJ (2002) Diaphragm defects occur in a CDH hernia model independently of myogenesis and lung formation. Am J Physiol Lung Cell Mol Physiol 283:L1310–L1314PubMed
Metadata
Title
The role of primary myogenic regulatory factors in the developing diaphragmatic muscle in the nitrofen-induced diaphragmatic hernia
Authors
Jens Dingemann
Takashi Doi
Elke Ruttenstock
Prem Puri
Publication date
01-06-2011
Publisher
Springer-Verlag
Published in
Pediatric Surgery International / Issue 6/2011
Print ISSN: 0179-0358
Electronic ISSN: 1437-9813
DOI
https://doi.org/10.1007/s00383-010-2834-8

Other articles of this Issue 6/2011

Pediatric Surgery International 6/2011 Go to the issue