Biochemical and Biophysical Research Communications
Identification of gene expression modifications in myostatin-stimulated myoblasts
Section snippets
Materials and methods
Cell culture and treatment. Chicken fetal myoblasts (CFMs) were prepared from the pectoralis muscles of 10-day-old White Leghorn chicken embryos. Briefly, the pectoralis muscles were removed and dissected in order to remove bone and skin. The muscle tissue was minced and incubated in growth medium, GM (DMEM supplemented with 10% fetal bovine serum, 2% chicken serum, and penicillin/streptomycin) at 37 °C in a 5% CO2 atmosphere, with intermittent shaking for 20 min. The dissociated cells were
Identification of expression-altered genes in myostatin stimulated CFMs
It has been reported that the mature portion of endogenous myostatin localized in the C-terminus from amino acid 266 to 376 [18]. Therefore, we expressed and purified the C-terminal truncated recombinant myostatin (Lys193-Ser375) as described [22]. To verify the biological function of the recombinant protein, a cytometry analysis and Northern blot were performed to determine CFMs responses to myostatin stimulation. Myostatin efficiently induced CFMs cell cycle arrest at G1 phase and decreased
Discussion
As a growth and differentiation factor, myostatin has been shown to determine muscle size by controlling its growth. However, molecular mechanism of myostatin action still remains to be explicated. The identification of gene expression alteration in response to myostatin stimulation could provide important information for understanding the molecular mechanisms of myostatin function for controlling skeletal muscle size. In the present study, we identified a number of genes whose expression was
Acknowledgments
This research was supported by the following grants: The National High Technology Research and Development Program of China (“863” Program, No. 2001AA222031), and the National Science Fund for Distinguished Young Scholars of China (No. 300250). We thank Sara Zhu for assistance with preparation of the manuscript.
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These authors contributed equally.