Suppression of myostatin with vector-based RNA interference causes a double-muscle effect in transgenic zebrafish.
Myostatin belongs to the transforming growth factor (TGF)-beta superfamily and is a potent negative regulator of skeletal muscle development and growth. We utilized microinjection of an antisense RNA-expressing vector to establish a hereditarily stable myostatin gene knockdown zebrafish strain with a double-muscle phenotype. Real-time PCR and immunostaining revealed that the myostatin messenger (m)RNA and protein levels in homozygous transgenic zebrafish were 33% and 26% those of the non-transgenic controls, respectively. Also, the mRNA levels of myogenic regulatory factor markers such as MyoD, myogenin, Mrf4, and Myf5 were dramatically elevated in myostatin-suppressed transgenic fish compared to the non-transgenic controls. Although there was no significant difference in body length, homozygous transgenic zebrafish were 45% heavier than non-transgenic controls. Histochemical analysis showed that the cross-sectional area of the muscle fiber of homozygous transgenic fish was twice as large as that of non-transgenic controls. This is the first model zebrafish with a hereditarily stable myostatin-suppressed genotype and a double-muscle phenotype.
Lee
Chiou-Yueh
CY
Institute of Microbiology and Biochemistry, National Taiwan University, Taipei, ROC.
Hu
Shao-Yang
SY
Gong
Hong-Yi
HY
Chen
Mark Hung-Chih
MH
Lu
Jenn-Kan
JK
Wu
Jen-Leih
JL
D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology* / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D023421:Models, Animal; D024510:Muscle Development; D009132:Muscles / Q000196:embryology*; D055435:Myostatin / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics; D029961:Zebrafish Proteins / Q000235:genetics*
C525914:Mstnb protein, zebrafish; D055435:Myostatin; D034741:RNA, Small Interfering; D029961:Zebrafish Proteins