Ali Javadmanesh; Mitra Riasi
Abstract
TGF-β group member myostatin (MSTN) inhibits the growth and differentiation of skeletal muscle. It is unknown that how total inhibition of MSTN restricts the hyperproliferation of muscle cells, though. This study aimed to determine the differentially expressed genes and biological pathways associated ...
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TGF-β group member myostatin (MSTN) inhibits the growth and differentiation of skeletal muscle. It is unknown that how total inhibition of MSTN restricts the hyperproliferation of muscle cells, though. This study aimed to determine the differentially expressed genes and biological pathways associated with myostatin in mouse C2C12 myogenic cells and skeletal muscle tissue utilizing high throughput mRNA expression data. In both tissue and cell line, two experimental groups were compared including wild-type vs. MSTN-Knockout. Transcriptome data were extracted from GEO and analyzed using the LIMMA package in R environment, and GEO2R. Significant differentially expressed genes were considered as False discovery rate < 0.05 with log fold change > |0.5|. Lastly, the possible biological pathways were examined with KEGG database and the protein-protein interaction (PPI) network was created using Cytoscape software. To find key genes in the PPI network, a topological analysis was conducted using the Network analyzer tool in Cytoscape software. The total number of differentially expressed genes was 14549 for C2C12 cell line and 45267 for mouse skeletal muscle tissue; among them 235 and 1425 transcripts were significant. The comparison between these two DEG lists showed that 49 genes were common between myogenic C2C12 cell and skeletal muscle tissue. Additionally, three biological pathways between the C2C12 cell line and the skeletal muscle tissue were in common and they were associated to the decrease of MSTN gene expression including: hypertrophic cardiomyopathy, Axon guidance, and dilated cardiomyopathy. These pathways were all related to muscle tissue. Despite the large number of DEGs, only 49 were in common in tissue and cell line; this could indicate that comparing a tissue with its relevant cell line at transcriptome level might not be precise enough to draw a solid conclusion. This is due to the nature of cell heterogeneity of most tissues including skeletal muscle.
Ali Javadmanesh; Amir Rashid Lamir; Mitra Riasi; Mehrdad Movahed Nasab; Nazila Dardmeh; Helia Khayyami; Kasra Khayami; Elnaz Karbaschian
Abstract
The growth and development of skeletal muscle tissue is largely regulated by myostatin during tissue development in embryos. This tissue may overgrow if myostatin expression is deficient. Gene expression may be regulated in a particular way by oligonucleotide antisense molecules. It has been demonstrated ...
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The growth and development of skeletal muscle tissue is largely regulated by myostatin during tissue development in embryos. This tissue may overgrow if myostatin expression is deficient. Gene expression may be regulated in a particular way by oligonucleotide antisense molecules. It has been demonstrated that a new DNA-based oligonucleotide can downregulate myostatin expression in a rat model. The purpose of this work was to evaluate the impact of a DNAi-based myostatin inhibitor on the visceral fat and leg muscle weights of Wistar rats undergoing strength training. Three groups of male rats, with an average weight of 203g ± 10.5, were chosen at four weeks of age. These cohorts comprised: 1) DNAi group had resistance training in addition to receiving 10 mg/kg of rat body weight of DNAi. 2) Resistance exercise and saline injection group. Group for injection of saline. Then, weight measurements for the carcass, heart, liver, left kidney, right kidney, spleen, visceral fat, twin muscles, soleus muscle, and left leg were made for each group. Histological assessment of the soleus muscle section was performed. One-way ANOVA was then used to examine the results, and means were compared using Tukey’s test. As the data show, the proposed molecule did not significantly contribute to an increase in body weight, in contrast to previous assumptions. Nonetheless, the twin muscles' relative and absolute weights increased significantly with visceral fat decreased with DNAi injection (P<0.05). Although weekly body weight increase and the final weights were not affected by DNAi injection, this could be explained by the loss of fat tissue during the experiment. This molecule is promising in increasing muscle tissue growth; however, further prolonged experiments and evaluating myostatin gene expression are recommended in future experiments.
Mitra Riasi; Elnaz Karbaschian; Ali Javadmanesh
Abstract
DNA-based approaches can now be utilized as low-risk methods to change gene expression. It appears that this approach has the ability to partially replace RNA-based approaches for altering gene expression, which in the majority of cases leads to immunological responses in patients. When utilized as a ...
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DNA-based approaches can now be utilized as low-risk methods to change gene expression. It appears that this approach has the ability to partially replace RNA-based approaches for altering gene expression, which in the majority of cases leads to immunological responses in patients. When utilized as a technique to silence target gene expression, DNA interference (DNAi) is a single-stranded DNA created to complement the upstream region of a gene. This DNAi molecule is stabilized using a variety of chemical changes, including phosphorothioates, methylphosphonate setC, etc. Several studies of the efficient application of DNA-based methods both in eukaryotic cell lines and the therapy of various disorders, such as Duchenne muscular dystrophy, cancer, etc., have been mentioned. Understanding the DNAi process, its transfer carriers, stabilization techniques, and their limitations is crucial for advancing these applications and predicting the future of DNAi both in basic science and the treatment of disorders brought on by abnormal gene expression. The main purpose of this review is introducing benefits of using DNAi in gene silencing. this review has discussed about different applications of DNAi in drug discovery and treatment, criteria of designing DNAi, possible modifications, introducing different types of carriers and limitations of DNAi administration.
