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KMID : 1025520090510060459
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Journal of Animal Science and Technology
2009 Volume.51 No. 6 p.459 ~ p.470
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Differential Proteome Expression of In vitro Proliferating Bovine Satellite Cells from Longissimus Dorsi, Deep Pectoral and Semitendinosus Muscle Depots in Response to Hormone Deprivation and Addition.
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Ramanna Valmiki Rajesh
Kim Seong-Kon
Park Mi-Rim
Park Min-Ah
Jang Eun-Joung
Hong Seung-Go
Jang Jong-Soo
Yoon Du-Hak
Kim Tae-Hun
Lee Hyun-Jeong
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Abstract
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The aim of this study was to analyze the proteome of proliferating bovine satellite cells from longissimus dorsi, deep pectoral and semitendinosus muscle depots which had been subjected to hormonal deprivation or addition in culture. For hormone deprivation or addition studies, the cells were either grown in 10% charcoal-dextran stripped fetal bovine serum(CD-FBS) or in 10% FBS supplemented medium. Further to analyze the effect of insulin like growth factor(IGF-1) and testosterone(TS), the cells were grown in 10% CD-FBS containing IGF-1(10ng/ml) or TS(10nM). Results have shown that hormone deprivation had a negative impact on proliferation of the cells from each of the muscle depots. In case of IGF-1 and TS addition, the proliferation levels were low compared with that of the cells grown in 10% FBS. Hence, to gain the insights of the proteins that are involved in such divergent levels of proliferation, the proteome of such satellite cells proliferating under the above mentioned conditions were analyzed using 2D-DIGE and MALDI-ToF/ToF. Thirteen proteins during hormone deprivation and nine proteins from hormone addition were found to be differentially expressed in all the cultures of the cells from the three depots. Moreover, the results highlighted in this study offer a role for each differentially expressed protein with respect to its effect on positive or negative regulation of cell proliferation.
Skeletal muscle is a tissue of major economic importance for meat production. Muscle fiber growth, along with the hormones which affect the growth of muscle tissue are important for meat industry(Campion, 1984). The number of muscle fibers in meat-producing animals is essentially fixed at birth, thus postnatal muscle growth results from hypertrophy of existing muscle fibers (Kamanga-Sollo et al, 2008). This fiber hypertrophy requires an increase in the number of myonuclei present in the fibers. However, the nuclei present in muscle fibers are unable to divide, so the nuclei must come from outside the fiber(Campion, 1984). The source of the nuclei needed to support fiber hyperplasia (cell proliferation) is mononucleated myogenic cells, satellite cells, located on the periphery of the fiber (Moss and Leblond, 1971). Muscle satellite cells play a crucial role in the postnatal muscle growth by fusing with existing muscle fibers and providing the nuclei required for postnatal fiber growth (Campion, 1984 Moss and Leblond, 1971).
A better understanding of the involvement of satellite cells in postnatal muscle hyperplasia represents a powerful key to improve the efficiency of muscle growth in meat-producing animals. Nevertheless, regulation of satellite cell activity in vivo is complex. Satellite cells may be regulated through extrinsic factors such as the innervation, hormonal signal, or locally produced growth factors(Allen and Rankin, 1990 Dodson et al, 1996). Among growth factors are the fibroblast growth factors (FGFs), insulin-like growth factors (IGFs), transforming growth factor b (TGFb), epidermal growth factor (EGF), platelet-derived growth factor (PDGF), or hepatocyte growth factor (HGF) (Allen et al, 1995). Even though satellite cells are regulated through diffusible factors arising from the fibers rather than through circulating factors, satellite cells may also be activated under nutritional (Hansen-Smith et al, 1979) or hormonal manipulation (McCuscker and Campion, 1986; Thompson et al, 1989). There are few reports about hormonal regulation of meat-animal satellite cells. Several hormones have been shown to affect the proliferation and differentiation of both primary satellite cells from bovine (Kamanga-Sollo et al, 2008) and rat (Allen and Boxhorn, 1987), along with myogenic cell lines(Florini and Magri, 1989). Androgenic and estrogenic implants are widely used, both individually and in combination, as growth promoters for cattle (Hayden et al, 1993; Johnson et al, 1996). Also, it was shown that artificial implantation of growth hormones increased the number of satellite cells (Johnson et al, 1998). Additionally, a number of studies in humans and animals have shown that hormone treatment increases the muscle fiber diameter. The number of myonuclei present in the muscle fibers was also shown to be increased (Bhasin et al, 2001; Sinha-Hikim et al, 2002, 2003). Also, it was shown that muscle satellite cells were subjected to apoptosis during serum deprivation (Mampuru et al, 1996), but all of these studies lack proteomic approach to elucidate the proteins that are involved in such a cell death process. Hence, the aim of this work was to look for muscle depot related differences for differentially expressed proteins in relation to proliferation under hormone deprivation and addition conditions. So, this study detail the comparison of growth rate and DIGE(Fluorescence Difference Gel Electrophoresis) dependent proteomic characterization during hormone deprivation and addition on proliferating satellite cells from bovine longissimus dorsi(LD), deep pectoral(DP) and semitendinosus (ST) muscle depots.
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KEYWORD
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Satellite cells, Depot, Proliferation, Hormone
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