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Functional Profiles of Growth Related Genes during Embryogenesis and Postnatal Development of Chicken and Mouse Skeletal Muscle Hakan Kocamis Myostatin (also known as growth differentiation factor/8), a recently identified member of the TGF-β family, has been shown to negatively regulate skeletal muscle growth. Activins, also members of the TGF-β family, and their binding protein, follistatin, once thought to be restricted to reproductive cycle function are in fact involved in the development of a wide variety of embryological and adult tissues, particularly skeletal muscles. Reverse-transcription ploymerase chain reaction (RT-PCR) was performed to measure the ontogeny of myostatin, activin-B, and follistatin gene expression during chicken embryonic development. Strong myostatin expression was found in the early chicken embryos (E 0, E 1) and the developmental expression pattern of myostatin mRNA coincided with the periods of primary and secondary muscle fiber formation. Follistatin transcripts followed a linear expression pattern from E 0 to E 20, while activin-B had a quadratic pattern. The ontogeny of myostatin gene expression was nearly identical in satellite cells isolated from pectoralis major (PM) and biceps femoris (BF) muscles of chicken. Activin-B mRNA level in PM satellite cells was higher than in BF satellite cells at 72 h and 120 h (P < 0.01), whereas levels in BF satellite cells were higher than in PM satellite cells at 96 h and 144 h (P < 0.01). Amounts of follistatin mRNA in PM satellite cells were higher than in BF satellite cells at 24, 96, and 120 h of culture (P < 0.01). No IGF-I gene expression was detected in either cell culture at any time point in the present study. IGF-II mRNA level plateaued in PM satellite cells by 48 h after plating (P < 0.05), and remained elevated until 144 h of culture. In ovo administration of rhIGF-I at E 3 altered myostatin, follistatin, activin-B, and TGF-β2 gene expressions during chicken embryonic development with emphasis on skeletal muscle development. Myostatin mRNA from pectoralis muscles of rhIGF-I injected embryos increased on E 10 (~ 2.5 fold) and remained high through E 13, whereas mRNA from control pectoralis muscles increased at E 9 and remained high until E 12. IGF-I, -II and IGF receptor-I mRNA and protein levels were determined in a wide variety of myostatin knockout mice tissues. IGF-I mRNA levels were not different between control and knockout mice tissues, whereas levels for IGF-II were significantly higher in myostatin knockout mice kidney and soleus muscles than that of control mice (P < 0.01). IGF-Receptor-1 mRNA levels from control mice heart (P < 0.05) and kidney (P < 0.01) were significantly higher than that of myostatin knockout mice, while levels were lower in control mice pectoralis muscle than that of knockout mice (P < 0.01). The strongly IGF-II positive cells were more common in myostatin knockout mice and were seen in a few foci in control mice, while no consistent differences in IGF-II immunoreactivity were detected between the two groups of mice kidneys.

Functional profiles of growth-related genes during embryogenesis and postnatal development of chicken and mouse skeletal muscle