Hakan KOCAMIŞ, Nurhayat GÜLMEZ, Şahin ASLAN, Mümtaz NAZLI, Scott A. GAHR

Alterations of Myostatin and Follistatin Gene Expression Profiles in Response to in ovo rhIGF-1 during Quail (Coturnix coturnix) Embryonic Development

Myostatin, along with its binding protein follistatin, have been demonstrated to inhibit skeletal muscle development during embryonic and postnatal development, whereas IGF-1 has been reported to stimulate skeletal muscle development and overall growth. The objectives of the current study were to determine the expression profiles of myostatin and follistatin genes, and to evaluate the impact of in ovo administration of recombinant human insulin-like growth factor-1 (rhIGF-1) on these gene expression patterns during quail embryogenesis, with an emphasis on skeletal muscle development. Quail eggs were injected once with 100 ng rhIGF-1 in 10 mM acetic acid, 0.1% BSA per embryo on day 3 of embryonic development. Total RNA was isolated from whole embryos on each of embryonic days (E) 3 to 6 (n = 6 per day/per treatment), from thoracic/abdominal halves of the embryos on E 7 and E 8 (n = 6 per day/per treatment), and from pectoralis muscle tissues on Es 9 to 16 (n= 4 per day/per treatment). Reverse-transcription polymerase chain reaction (RT-PCR) was used to determine the relative expression pattern. Myostatin mRNA was first detected on E 5, while follistatin mRNA was first detected on E 3. Although myostatin mRNA levels from the control group declined from E 10 to E 12, they increased from E 9 to E16; on E 16 it reached its highest point in the rhIGF-1 injected group. In ovo administration of rhIGF-1 on E 3 appeared to delay myostatin expression approximately 1 day as compared to control embryos at E 9, while it enhanced follistatin expression on E 9. Our results suggest that myostatin and follistatin genes may be pivotal regulators of skeletal muscle growth, and that in ovo administration of rhIGF-1on E 3 may alter the expression profiles of myostatin and follistatin genes, which might enhance the skeletal muscle mass of quail embryos.

Anahtar Kelimeler:

Myostatin, follistatin, quail embryos, in ovo rhIGF-1

Alterations of Myostatin and Follistatin Gene Expression Profiles in Response to in ovo rhIGF-1 during Quail (Coturnix coturnix) Embryonic Development

Keywords:

Myostatin, follistatin, quail embryos, in ovo rhIGF-1,

PDF

___

Kocamis, H., Kirkpatrick-Keller, D.C., Richter, J., Killefer, J.: The ontogeny of myostatin, follistatin and activin-B mRNA expression during chicken embryonic development. Growth Dev. Aging, 1999; 63: 143-150. Kocamis, H., Gahr, S.A., Richter, J., Kirkpatrick-Keller, D.C., Killefer, J.: Myostatin and TGF-β2 gene expression patterns in response to in ovo administration of rhIGF-1 during chicken embryonic development. Growth Dev. Aging, 2002; 66: 3-10.
Chomczynski, P., Sacci, N.: Single step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal. Biochem., 1987; 162: 156-159.
Fredette, B.J., Landmesser, L.T.: Relationship of primary and secondary myogenesis to fiber type development in embryonic chick muscle. Dev. Biol. 1991; 143: 1-18.
Graham, A., Lumbsden, A.: Interactions between rhombomeres modulate Krox-20 and follistatin expression in the chick embryo hindbrain. Development, 1996; 122: 473-480.
Davis, A.J., Johnson, P.A.: Expression pattern of messenger ribonucleic acid for follistatin and the inhibin/activin subunits during follicular and testicular development in Gallus domesticus. Biol. Reprod., 1998; 59: 271-277.
Yamamura, M., Uyemura, K., Deans, R.J., Weinberg, K., Rea, T.H., Bloom, B.R., Modlin, R.L.: Defining protective responses to pathogens: cytokine profiles in lepropsy lesions. Science, 1991; 254: 277-279.
Ji, S., Losinski, R.L., Cornelius, S.G., Frank, G.R., Willis, G.M., Gerrand, D.E., Depreux, F.F.S., Spurlock, M.E.: Myostatin expression in porcine tissues: tissue specificity and developmental and postnatal regulation. Am. J. Physiol., 1998; 275: 1265- 1273.
Florini, J.R., Ewton, D.Z., Roof, S.L.: IGF-I stimulates terminal myogenic differentiation by induction of myogenin gene expression. Mol. Endocrinol., 1991; 5: 718-724.
Li, H., Capetanaki, Y.: An E box in the desmin promoter cooperates with the E box and MEF-2 sites of a distal enhancer to direct muscle-specific transcription. EMBO J., 1994; 13: 3580- 3589.
Ferrell, E.R., Conte, V., Lawrence, E.C., Roth, S.M., Hagberg, J.M., Hurley, B.F.: Frequent sequence variation in the human myostatin (GDF8) gene as a marker for analysis of muscle-related phenotypes. Genomics, 1999; 62: 203-207.