Analysis of myostatin gene structure, expression and function in zebrafish.
Myostatin is a member of the TGF-beta family that functions as a negative regulator of skeletal muscle development and growth in mammals. Recently, Myostatin has also been identified in fish; however, its role in fish muscle development and growth remains unknown. We have reported here the isolation and characterization of myostatin genomic gene from zebrafish and analysis of its expression in zebrafish embryos, larvae and adult skeletal muscles. Our data showed that myostatin was weakly expressed in early stage zebrafish embryos, and strongly expressed in swimming larvae, juvenile and skeletal muscles of adult zebrafish. Transient expression analysis revealed that the 1.2 kb zebrafish myostatin 5' flanking sequence could direct green fluorescent protein (GFP) expression predominantly in muscle cells, suggesting that the myostatin 5' flanking sequence contained regulatory elements required for muscle expression. To determine the biological function of Myostatin in fish, we generated a transgenic line that overexpresses the Myostatin prodomain in zebrafish skeletal muscles using a muscle-specific promoter. The Myostatin prodomain could act as a dominant negative and inhibit Myostatin function in skeletal muscles. Transgenic zebrafish expressing the Myostatin prodomain exhibited no significant change in myogenic gene expression and differentiation of slow and fast muscle cells at their embryonic stage. The transgenic fish, however, exhibited an increased number of myofibers in skeletal muscles, but no significant difference in fiber size. Together, these data demonstrate that Myostatin plays an inhibitory role in hyperplastic muscle growth in zebrafish.
Xu
Cheng
C
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, MD 21202, USA.
Wu
Gang
G
Zohar
Yonathan
Y
Du
Shao-Jun
SJ
D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D001835:Body Weight; D002874:Chromosome Mapping; D017931:DNA Primers; D004625:Embryo, Nonmammalian / Q000378:metabolism; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D017403:In Situ Hybridization; D007814:Larva / Q000378:metabolism; D008164:Luminescent Proteins; D008845:Microinjections; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000502:physiology; D055435:Myostatin; D010957:Plasmids / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D016212:Transforming Growth Factor beta / Q000235:genetics*; D019076:Transgenes; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D029961:Zebrafish Proteins
D017931:DNA Primers; D008164:Luminescent Proteins; C525914:Mstnb protein, zebrafish; D055435:Myostatin; D016212:Transforming Growth Factor beta; D029961:Zebrafish Proteins; D049452:Green Fluorescent Proteins