TY - JOUR
T1 - The role of primary myogenic regulatory factors in the developing diaphragmatic muscle in the nitrofen-induced diaphragmatic hernia
AU - Dingemann, Jens
AU - Doi, Takashi
AU - Ruttenstock, Elke
AU - Puri, Prem
PY - 2011/6
Y1 - 2011/6
N2 - 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 microdissected and divided into CDH group and controls. MyoD and Myf5 mRNA-expression were determined using Realtime 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.
AB - 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 microdissected and divided into CDH group and controls. MyoD and Myf5 mRNA-expression were determined using Realtime 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.
KW - Congenital diaphragmatic hernia
KW - Diaphragmatic defect, MyoD, Myf5
KW - Nitrofen
UR - http://www.scopus.com/inward/record.url?scp=79960007373&partnerID=8YFLogxK
U2 - 10.1007/s00383-010-2834-8
DO - 10.1007/s00383-010-2834-8
M3 - Article
C2 - 21258934
AN - SCOPUS:79960007373
SN - 0179-0358
VL - 27
SP - 579
EP - 582
JO - Pediatric Surgery International
JF - Pediatric Surgery International
IS - 6
ER -