Transgenic salmon overexpressing growth hormone exhibit decreased myostatin transcript and protein expression.
To characterize the role of myostatin (MSTN) in fast growing animals and to examine the relationship between MSTN and growth hormone (GH), MSTN transcript and protein expression were measured in coho salmon overexpressing GH and in wild-type coho salmon. Quantitative real-time RT-PCR and western analysis were used to measure RNA expression of the two salmonid MSTN genes (1 and 2) and levels of MSTN immunoreactive protein (MIP) in developing embryos and adult coho salmon tissues. In transgenic and control coho embryos, MSTN1 and MSTN2 RNA expression were initially observed at about the time of eying, and a 42 kDa MIP was just detected prior to hatching. Expression of the MSTN1 transcript in transgenic salmon was not different from that in wild-type adult coho salmon muscle and brain tissue. However, expression of the MSTN2 transcript was less in white muscle, and greater in red muscle, from transgenic fish compared to wild-type salmon of the same size. Northern analysis revealed that expression of the MSTN2 transcript was less in white muscle from wild-type, age-matched salmon than in transgenic fish. In addition, there was less presumed bioactive MIP in muscle taken from adult transgenic fish compared to controls and evidence of differential protein processing. Decreased MSTN expression in faster growing fish suggests that MSTN does act as a negative regulator of muscle growth in fish, as it does in mammals. The results of this study also suggest that the anabolic effects of GH could be mediated through MSTN.
Roberts
Steven B
SB
Marine Biological Laboratory, 7 MBL Street, Woods Hole, MA 02543, USA. sroberts@mbl.edu
McCauley
Linda A R
LA
Devlin
Robert H
RH
Goetz
Frederick W
FW
D000704:Analysis of Variance; D000818:Animals; D030801:Animals, Genetically Modified / Q000378:metabolism*; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D017931:DNA Primers; D015870:Gene Expression; D013006:Growth Hormone / Q000378:metabolism*; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000254:growth & development / Q000378:metabolism*; D055435:Myostatin; D017684:Oncorhynchus kisutch / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism*
D017931:DNA Primers; D055435:Myostatin; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D013006:Growth Hormone