Construction of a eukaryotic expression vector for pEGFP-FST and its biological activity in duck myoblasts

Li, Xinxin; Wang, Jiwen; Liu, Hehe; Wang, Haohan; Sun, Lin; Yang, Chao; Li, Liang; Zheng, Yi

doi:10.1016/j.ejbt.2014.07.002

Cited by 3 publications

(3 citation statements)

References 15 publications

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“…The BrdU assay was employed to analyse the proliferation states of the myoblasts, and the results showed that the percentage of BrdU-labelled nuclei was higher in the pEGFP-duFST group ( Figures 1 D– 1 E, P <0.05), indicating a positive role of FST on duck myoblast proliferation. In transient transfection assay, our previous research showed that pEGFP-N1 alone did not have significant effects on the myoblast proliferation, the myoblast vitality and the mRNA expression of FST [ 20 ]. Thus, we can exclude the potential influences of EGFP on the endpoint analysis.…”

Section: Resultsmentioning

confidence: 99%

Follistatin could promote the proliferation of duck primary myoblasts by activating PI3K/Akt/mTOR signalling

Liu

Wang

et al. 2014

Bioscience Reports

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FST (follistatin) is essential for skeletal muscle development, but the intracellular signalling networks that regulate FST-induced effects are not well defined. We sought to investigate whether FST promotes the proliferation of myoblasts through the PI3K (phosphoinositide 3-kinase)/Akt (protein kinase B)/mTOR (mammalian target of rapamycin) signalling. In the present study, we transfected the pEGFP-duFST plasmid and added PI3K and mTOR inhibitors to the medium of duck primary myoblasts. Then, we analysed the cellular phenotypic changes that occurred and analysed the expression of target genes. The results showed that FST promoted myoblast proliferation, induced the mRNA expression of PI3K, Akt, mTOR, 70-kDa ribosomal protein S6K (S6 kinase) and the protein expression of phospho-Akt (Thr308), mTOR, phospho-mTOR (serine 2448), phospho-S6K (Ser417), inhibited the mRNA expression of FoxO1, MuRF1 (muscle RING finger-1) and the protein expression of phospho-FoxO1 (Ser256). Moreover, we found that the overexpression of FST could alleviate the inhibitory effect of myoblast proliferation caused by the addition of LY294002, a PI3K inhibitor. Additionally, the overexpression of duck FST also relieved the inhibition of myoblast proliferation caused by the addition of rapamycin (an mTOR inhibitor) through PI3K/Akt/mTOR signalling. In light of the present results, we hypothesize that duck FST could promote myoblast proliferation, which is dependent on PI3K/Akt/mTOR signalling.

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Section: Resultsmentioning

confidence: 99%

Follistatin could promote the proliferation of duck primary myoblasts by activating PI3K/Akt/mTOR signalling

Liu

Wang

et al. 2014

Bioscience Reports

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“…Among the core genes that regulate muscle development, mstn is the most important negative regulator (Parsons et al 2006 ). Generally, mstn keeps muscle cells in G0/G1 and G2 phases by regulating the binding of Myog and Smad proteins, thereby restricting the differentiation and proliferation of myoblasts (Li et al 2014 ). Previous studies have shown that the regulation of mstn is a very complex process.…”

Section: Discussionmentioning

confidence: 99%

Transcriptional Knock-down of mstn Encoding Myostatin Improves Muscle Quality of Nile Tilapia (Oreochromis niloticus)

Wang,

Yan,

Tao

et al. 2023

Mar Biotechnol

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Myostatin (encoded by mstn) negatively regulates skeletal muscle mass and affects lipid metabolism. To explore the regulatory effects of mstn on muscle development and lipid metabolism in Nile tilapia (Oreochromis niloticus), we used antisense RNA to transcriptionally knock-down mstn. At 180 days, the body weight and body length were significantly higher in the mstn-knock-down group than in the control group (p < 0.05). Additionally, fish with mstn-knock-down exhibited myofiber hyperplasia but not hypertrophy. Oil red O staining revealed a remarkable increase in the area of lipid droplets in muscle in the mstn-knockdown group (p < 0.05). Nutrient composition analyses of muscle tissue showed that the crude fat content was significantly increased in the mstn-knock-down group (p < 0.05). The contents of saturated fatty acids, monounsaturated fatty acids, and polyunsaturated fatty acids were all significantly increased in the mstn-knock-down group (p < 0.05). Comparative transcriptome analyses revealed 2420 significant differentially expressed genes between the mstn-knock-down group and the control group. KEGG analysis indicates that disruptions to fatty acid degradation, glycerolipid metabolism, and the PPAR signaling pathway affect muscle development and lipid metabolism in mstn-knock-down Nile tilapia: acaa2, eci1, and lepr were remarkably up-regulated, and acadvl, lpl, foxo3, myod1, myog, and myf5 were significantly down-regulated (p < 0.05). These results show that knock-down of mstn results in abnormal lipid metabolism, acceleration of skeletal muscle development, and increased adipogenesis and weight gain in Nile tilapia.

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“…Among the core genes that regulate muscle development, mstn is the most important negative regulator (Parsons et al 2006). Generally, mstn keeps muscle cells in G0/G1 and G2 phases by regulating the binding of Myog and Smad proteins, thereby restricting the differentiation and proliferation of myoblasts (Li et al 2014). Previous studies have shown that the regulation of mstn is a very complex process.…”

Section: Molecular Mechanism Which Mstn Affects and Lipid Metabolismmentioning

confidence: 99%

Transcriptional knock-down of mstn encoding myostatin improves muscle quality of Nile tilapia (Oreochromis niloticus)

Wang

Yan

Tao

et al. 2023

Preprint

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Background Myostatin (encoded by mstn) negatively regulates skeletal muscle mass and affects lipid metabolism. Relieving the inhibitory effect of mstn on growth can improve the muscle yield of teleost fishes. To explore the regulatory effects of mstn on muscle development and lipid metabolism in Nile tilapia (Oreochromis niloticus), we used antisense RNA to transcriptionally knock-down mstn. At 180 d, the body weight and body length were significantly higher in the mstn-knock-down group than in the control group (p < 0.05). Hematoxylin–eosin staining revealed that fish in the mstn-knock-down group exhibited myofiber hyperplasia but not hypertrophy. Oil red O staining revealed a remarkable increase in the area of lipid droplets in muscle in the mstn-knock-down group compared with that in the control group (p < 0.05). Nutrient composition analyses of muscle tissue showed that the crude fat content was significantly increased in the mstn-knock-down group (p < 0.05). The contents of saturated fatty acids, monounsaturated fatty acids, and polyunsaturated fatty acids were all significantly increased in the mstn-knock-down group (p < 0.05). Comparative transcriptome analyses revealed 2,420 significant differentially expressed genes between the mstn-knock-down group and the control group, including 1,055 up-regulated genes and 1,365 down-regulated genes. Among them, genes related to myoblast differentiation and lipid metabolism were enriched in fatty acid degradation, glycerolipid metabolism, and peroxisome proliferator activated receptor (PPAR) signaling pathways. The accuracy of the RNA-seq data was confirmed by qRT-PCR analyses. Our results indicate that disruptions to fatty acid degradation, glycerolipid metabolism, and the PPAR signaling pathway affect muscle development and lipid metabolism in mstn-knock-down Nile tilapia: acaa2, eci1, and lepr were remarkably up-regulated, and acadvl, lpl, foxo3, myod1, myog, and myf5 were significantly down-regulated (p < 0.05). These results show that knock-down of mstn results in abnormal lipid metabolism, acceleration of skeletal muscle development, and increased adipogenesis and weight gain in Nile tilapia.

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Construction of a eukaryotic expression vector for pEGFP-FST and its biological activity in duck myoblasts

Cited by 3 publications

References 15 publications

Follistatin could promote the proliferation of duck primary myoblasts by activating PI3K/Akt/mTOR signalling

Follistatin could promote the proliferation of duck primary myoblasts by activating PI3K/Akt/mTOR signalling

Transcriptional Knock-down of mstn Encoding Myostatin Improves Muscle Quality of Nile Tilapia (Oreochromis niloticus)

Transcriptional knock-down of mstn encoding myostatin improves muscle quality of Nile tilapia (Oreochromis niloticus)

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