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Find video protocols related to scientific articles indexed in Pubmed.
Molecular cloning and expression pattern of duck Six1 and its preliminary functional analysis in myoblasts transfected with eukaryotic expression vector.
Indian J. Biochem. Biophys.
PUBLISHED: 10-10-2014
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Skeletal muscle development is regulated by Six1, an important myogenic transcription factor. However, the functional analysis of duck Six1 has not been reported. Here, we cloned the coding domain sequence (CDS) region of the duck Six1 gene using RT-PCR and RACE methods. Bioinformatics analysis revealed that duck Six1 CDS region comprised of 849 bp and encoded 282 amino acids and had a high degree of homology with other species, suggesting that the functions of duck Six1 gene are conserved among other animals. Real-time PCR used to determine the mRNA expression profiles of duck Six1 in different tissues and different developmental stages showed that Six1 was highly expressed in skeletal muscle and the embryonic stage. Furthermore, the eukaryotic expression vector pEGFP-duSix1 was constructed and transfected into the duck myoblasts; the MTT assay revealed an obvious increase of cell proliferation after transfection. The expression profiles of Six1, Myf5 and MyoD showed that their expression levels were significantly increased. These results together suggested that pEGFP-duSix1 vector was constructed successfully and overexpression of duck Six1 in the myoblasts could promote cell proliferation activity and significant up-regulate expression of Myf5 and MyoD.
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Follistatin could promote the proliferation of duck primary myoblasts by activating PI3K/Akt/mTOR signalling.
Biosci. Rep.
PUBLISHED: 09-10-2014
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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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Effects of the regulation of follistatin mRNA expression by IGF-1 in duck (Anas platyrhynchos) skeletal muscle.
Growth Horm. IGF Res.
PUBLISHED: 01-17-2014
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The IGF-1 and TGF-? pathways have been shown to be involved in regulating muscle development. Many mediators that are associated with the regulation of muscle development have been found to participate in the cross-talk between these two pathways. To research the relationships between IGF-1 and the follistatin-mediated TGF-? pathways in duck skeletal muscle development, a series of studies were conducted. The results showed that follistatin had similar expression patterns to IGF-1 during duck embryonic muscle development. The in ovo feeding of IGF-1 to duck eggs was shown to increase follistatin expression in the duck skeletal muscle. Thus, IGF-1 may induce the mRNA expression of follistatin. These results suggest that follistatin may be a key regulator of multiple signaling cascades responding to the cross-talk between the IGF-1 and TGF-? pathways.
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The effects of endoplasmic reticulum stress response on duck decorin stimulate myotube hypertrophy in myoblasts.
Mol. Cell. Biochem.
PUBLISHED: 01-30-2013
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Decorin inhibits the transforming growth factor superfamily to stimulate skeletal muscle hypertrophy. The endoplasmic reticulum (ER) plays a crucial role in the synthesis and folding of proteins. The disruption of ER homeostasis affects protein folding and causes ER stress, which is associated with protein synthesis in muscle cells. The purpose of this study was to establish the mechanism by which decorin promotes myoblast proliferation and myotube hypertrophy as mediated by an ER stress-related pathway. Duck myoblasts were incubated for 24 h with tunicamycin. Our results show that tunicamycin triggered ER stress in myoblasts and led to an increase in protein synthesis via a decorin-dependent pathway. We then transfected the decorin gene into duck myoblasts in vitro, and the results demonstrated that overexpression of duck decorin increased myogenic determination factor and follistatin expression and decreased myogenin and myostatin expression. Moreover, the results demonstrated that overexpression of duck decorin led to higher expression of ER stress marker genes. This increased expression trend can be alleviated in vitro by 4-PBA, which is a chemical chaperone that inhibits palmitate-induced ER stress. Collectively, our data show that duck decorin promotes myoblast proliferation mediated by an ER stress-related pathway.
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What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

How does it work?

We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.