Molecular and functional characterization of the adiponectin (AdipoQ) gene in goat skeletal muscle satellite cells
2
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
Objective: It is commonly accepted that adiponectin binds to its two receptors to regulate fatty acid metabolism in adipocytes. To better understand their functions in the regulation of intramuscular adipogenesis in goats, we cloned the three genes (adiponectin [AdipoQ], adiponectin receptor 1 [AdipoR1], and AdipoR2) encoding these proteins and detected their mRNA distribution in different tissues. We also determined the role of AdipoQ in the adipogenic differentiation of goat skeletal muscle satellite cells (SMSCs). Methods: SMSCs were isolated using 1 mg/mL Pronase E from the longissimus dorsi muscles of 3-day-old female Nanjiang brown goats. Adipogenic differentiation was induced in satellite cells by transferring the cells to Dulbecco's modified Eagle's medium supplemented with an isobutylmethylxanthine, dexamethasone and insulin cocktail. The pEGFP-N1-AD plasmid was transfected into SMSCs using Lipofectamine 2000. Expression of adiponectin in tissues and SMSCs was detected by quantitative polymerase chain reaction and immunocytochemical staining. Results: The three genes were predominantly expressed in adipose and skeletal muscle tissues. According to fluorescence and immunocytochemical analyses, adiponectin protein expression was only observed in the cytoplasm, suggesting that adiponectin is localized to the cytoplasm of goat SMSCs. In SMSCs overexpressing the AdipoQ gene, adiponectin promoted SMSC differentiation into adipocytes and significantly (p<0.05) up-regulated expression of AdipoR2, acetyl-CoA carboxylase, fatty-acid synthase, and sterol regulatory element-binding protein-1, though expression of CCAAT/enhancer-binding protein-α, peroxisome proliferator-activated receptor γ, and AdipoR1 did not change significantly. Conclusion: Adiponectin induced SMSC differentiation into adipocytes, indicating that adiponectin may promote intramuscular adipogenesis in goat SMSC.
Abstract
Objective: It is commonly accepted that adiponectin binds to its two receptors to regulate fatty acid metabolism in adipocytes. To better understand their functions in the regulation of intramuscular adipogenesis in goats, we cloned the three genes (adiponectin [AdipoQ], adiponectin receptor 1 [AdipoR1], and AdipoR2) encoding these proteins and detected their mRNA distribution in different tissues. We also determined the role of AdipoQ in the adipogenic differentiation of goat skeletal muscle satellite cells (SMSCs). Methods: SMSCs were isolated using 1 mg/mL Pronase E from the longissimus dorsi muscles of 3-day-old female Nanjiang brown goats. Adipogenic differentiation was induced in satellite cells by transferring the cells to Dulbecco's modified Eagle's medium supplemented with an isobutylmethylxanthine, dexamethasone and insulin cocktail. The pEGFP-N1-AD plasmid was transfected into SMSCs using Lipofectamine 2000. Expression of adiponectin in tissues and SMSCs was detected by quantitative polymerase chain reaction and immunocytochemical staining. Results: The three genes were predominantly expressed in adipose and skeletal muscle tissues. According to fluorescence and immunocytochemical analyses, adiponectin protein expression was only observed in the cytoplasm, suggesting that adiponectin is localized to the cytoplasm of goat SMSCs. In SMSCs overexpressing the AdipoQ gene, adiponectin promoted SMSC differentiation into adipocytes and significantly (p<0.05) up-regulated expression of AdipoR2, acetyl-CoA carboxylase, fatty-acid synthase, and sterol regulatory element-binding protein-1, though expression of CCAAT/enhancer-binding protein-α, peroxisome proliferator-activated receptor γ, and AdipoR1 did not change significantly. Conclusion: Adiponectin induced SMSC differentiation into adipocytes, indicating that adiponectin may promote intramuscular adipogenesis in goat SMSC.
참고문헌 (30)
- Scherer PE, "A novel serum protein similar to C1q, produced exclusively in adipocytes", J Biol Chem, 270, pp.26746~26749 (1995)[DOI]
- Katsiougiannis S, "Salivary gland epithelial cells: a new source of the immunoregulatory hormone adiponectin", Arthritis Rheum, 54, pp.2295~2299 (2006)[DOI]
- Staiger H, "Induction of adiponectin gene expression in human myotubes by an adiponectincontaining HEK293 cell culture supernatant", Diabetologia, 46, pp.956~960 (2003)[DOI]
- Berner HS, "Adiponectin and its receptors are expressed in bone-forming cells", Bone, 35, pp.842~849 (2004)[DOI]
- Fruebis J, "Proteolytic cleavage product of 30-kDa adipocyte complement-related protein increases fatty acid oxidation in muscle and causes weight loss in mice", Proc Natl Acad Sci USA, 98, pp.2005~2010 (2001)[DOI]
- Yamauchi T, "Cloning of adiponectin receptors that mediate antidiabetic metabolic effects", Nature, 423, pp.762~769 (2003)[DOI]
- Yamauchi T, "Targeted disruption of AdipoR1 and AdipoR2 causes abrogation of adiponectin binding and metabolic actions", Nat Med, 13, pp.332~339 (2007)[DOI]
- Fang X, "Leptin prevents the metabolic effects of adiponectin in L6 myotubes", Diabetologia, 52, pp.2190~2200 (2009)[DOI]
- Yoon MJ, "Adiponectin increases fatty acid oxidation in skeletal muscle cells by sequential activation of AMP-activated protein kinase, p38 mitogen-activated protein kinase, and peroxisome proliferator-activated receptor alpha", Diabetes, 55, pp.2562~2570 (2006)[DOI]
- Miaomiao Niu, "Adiponectin induced AMP-activated protein kinase impairment mediates insulin resistance in Bama mini-pig fed high-fat and high-sucrose diet", Animal Bioscience, 30(8), pp.1190~1197 (2017), 아세아·태평양축산학회[DOI]
- Shapiro L, "The crystal structure of a complement-1q family protein suggests an evolutionary link to tumor necrosis factor", Curr Biol, 8, pp.335~340 (1998)[DOI]
- Tomas E, "Enhanced muscle fat oxidation and glucose transport by ACRP30 globular domain: acetyl-CoA carboxylase inhibition and AMP-activated protein kinase activation", Proc Natl Acad Sci USA, 99, pp.16309~16313 (2002)[DOI]
- Iwabu M, "Adiponectin and AdipoR1 regulate PGC-1alpha and mitochondria by Ca(2+) and AMPK", SIRT1, 2010, pp.464, 1313~1319
- Fu Y, "Adiponectin promotes adipocyte differentiation, insulin sensitivity, and lipid accumulation", J Lipid Res, 46, pp.1369~1379 (2005)[DOI]
- Meng X, "Increasing fatty acid production in E. coli by simulating the lipid accumulation of oleaginous microorganisms", J Ind Microbiol Biotechnol, 38, pp.919~925 (2011)[DOI]
- Chakravarty B, "Human fatty acid synthase: structure and substrate selectivity of the thioesterase domain", Proc Natl Acad Sci USA, 101, pp.15567~15572 (2004)[DOI]
- White UA, "Transcriptional factors that promote formation of white adipose tissue", Mol Cell Endocrinol, 318, pp.10~14 (2010)[DOI]
- Taniguchi M, "Adipogenesis of bovine perimuscular preadipocytes", Biochem Biophys Res Commun, 366, pp.54~59 (2008)[DOI]
- Johnson PF, "Molecular stop signs: regulation of cell-cycle arrest by C", EBP transcription factors, 2005, pp.118, 2545~2555
- Lefterova MI, "PPARgamma and C", EBP factors orchestrate adipocyte biology via adjacent binding on a genome-wide scale, 2008, pp.22, 2941~2952
- Ren J, "DOG 1.0: illustrator of protein domain structures", Cell Res, 19, pp.271~273 (2009)[DOI]
- Robinson K, "Effect of statin therapy on plasma adiponectin concentrations in patients with the sepsis syndrome: a preliminary investigation", Intensive Care Med, 37, pp.1388~1389 (2011)[DOI]
- Shams M, "The relationship between serum adiponectin levels with the presence and severity of coronary artery disease", Arch Iran Med, 15, pp.611~616 (2012)
- Fiaschi T, "Globular adiponectin induces differentiation and fusion of skeletal muscle cells", Cell Res, 19, pp.584~597 (2009)[DOI]
- Narasimhan ML, "Osmotin is a homolog of mammalian adiponectin and controls apoptosis in yeast through a homolog of mammalian adiponectin receptor", Mol Cell, 17, pp.171~180 (2005)[DOI]
- Ramachandran R, "Molecular cloning and tissue expression of chicken AdipoR1 and AdipoR2 complementary deoxyribonucleic acids", Domest Anim Endocrinol, 33, pp.19~31 (2007)[DOI]
- Ding ST, "Cloning and expression of porcine adiponectin and adiponectin receptor 1 and 2 genes in pigs", J Anim Sci, 82, pp.3162~3174 (2004)[DOI]
- Korner A, "Adiponectin expression in humans is dependent on differentiation of adipocytes and down-regulated by humoral serum components of high molecular weight", Biochem Biophys Res Commun, 337, pp.540~550 (2005)[DOI]
- Kim CW, "Induced polymerization of mammalian acetyl-CoA carboxylase by MIG12 provides a tertiary level of regulation of fatty acid synthesis", Proc Natl Acad Sci USA, 107, pp.9626~9631 (2010)[DOI]
- Osborne TF, "Evolutionary conservation and adaptation in the mechanism that regulates SREBP action: what a long, strange tRIP it's been", Genes Dev, 23, pp.2578~2591 (2009)[DOI]