Dynamic expression of molecules that control limb muscle development including Fhl1 in hind limbs of different gestational age.
Muscle abnormality could be a key reason for congenital clubfoot (CCF) deformity, which manifests itself during fetal development. FHL1 down-regulated expression is involved in the formation of skeletal muscle abnormalities in CCF and FHL1 gene mutations contribute to the development of some kinds of myopathies. Therefore, detecting dynamic expression of Fhl1 and other molecules (Hgf, MyoD1, Myogenin, and Myh4) that control limb muscle development in hind limbs of different gestational age will provide a foundation for further research on the molecular mechanism involves in the myopathies or CCF. The dynamic gene expression levels of Fhl1, Hgf, MyoD1, Myogenin, and Myh4 in the lower limbs of E16, E17, E19, and E20 rat embryos were examined by real-time RT-PCR. Immunofluorescence was used to detect formation of specific muscle fibers (fast or slow fibers) in distal E17 hind limbs. The expression levels of Fhl1, Hgf, MyoD1, Myogenin, and Myh4 were varying in hind limbs of different gestational age. Real-time PCR results showed that all the genes that control skeletal muscle development except for Fhl1 exhibited a peak in E17 lower limbs. Immunofluorescence results showed obviously positive fast-myosin in the distal E17 lower limbs and meanwhile slow-myosin had no apparently signals. E17 was a critical time point for terminal skeletal muscle differentiation in the lower limbs of rat embryos.
Key laboratory of health ministry for congenital malformation, Shengjing Hospital, China Medical University, Shenyang, China.
Li-Li
LL
Wang
Zhao-Hong
ZH
Peng
Yang
Y
Fan
Lian-Yong
LY
Li
Di
D
Wu
Yi
Y
Zhang
Jia-Ning
JN
Miao
Yu-Zuo
YZ
Bai
Zheng-Wei
ZW
Yuan
Wei-Lin
WL
Wang
Kai-Lai
KL
Sun
D000818:Animals; D003025:Clubfoot / Q000235:genetics; D004622:Embryo, Mammalian / Q000002:abnormalities; D005260:Female; D005455:Fluorescent Antibody Technique; D005865:Gestational Age; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis; D006614:Hindlimb / Q000002:abnormalities* / Q000378:metabolism; D060588:LIM Domain Proteins / Q000096:biosynthesis* / Q000235:genetics; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D009124:Muscle Proteins / Q000096:biosynthesis* / Q000235:genetics; D018482:Muscle, Skeletal / Q000002:abnormalities* / Q000196:embryology; D009154:Mutation; D017570:MyoD Protein / Q000096:biosynthesis; D018008:Myogenin / Q000096:biosynthesis; D018995:Myosin Heavy Chains / Q000096:biosynthesis; D011247:Pregnancy; D051381:Rats
C517184:Fhl1 protein, rat; D060588:LIM Domain Proteins; D009124:Muscle Proteins; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D018008:Myogenin; D017228:Hepatocyte Growth Factor; D018995:Myosin Heavy Chains
Congenital clubfoot; Fhl1; dynamic expression; myopathy