Functional analyses of miRNA-146b-5p during myogenic proliferation and differentiation in chicken myoblasts

Abstract

Background: In the poultry and livestock industries, precise genetic information is crucial for improving economic traits. Thus, functional genomic studies help to generate faster, healthier, and more efficient animal production. Chicken myoblast cells, which are required for muscle development and regeneration, are particularly important because chicken growth is closely related to muscle mass. Results: In this study, we induced expression of microRNA-146b-5p mediated by the piggyBac transposon system in primary chicken myoblast (pCM) cells. Subsequently, we analyzed and compared the proliferation and differentiation capacity and also examined the expression of related genes in regular pCM (rpCM) cells and pCM cells overexpressing miRNA-146b-5p (pCM-146b OE cells). pCM-146b OE cells showed increased proliferation and upregulated gene expression related to cell proliferation. In addition, next-generation sequencing analyses were performed to compare global gene expression patterns between rpCM cells and pCM-146b OE cells. We found that the higher proliferation in pCM-146b OE cells was the result of upregulation of gene sets related to the cell cycle. Moreover, miRNA-146b-5p overexpression had inhibitory effects on myotube differentiation in pCM cells. Conclusions: Collectively these results demonstrate that miR-146b-5p is closely related to the proliferation and differentiation of chicken myogenic cells as a modulator of post-transcription. Background Since the genome sequences of avian species have become available, research has aimed to increase muscle mass, enhance muscle regeneration, and reduce fatty acid accumulation to improve growth. Understanding the genes or genetic markers involved in biological functions and regulatory pathways can help to improve economically important traits in the poultry industry [1-3]. Thus, functional genomic studies are powerful and effective for investigating the modulatory mechanisms between cell proliferation and differentiation, in particular in skeletal muscles [4-7]. Our study was conducted in chicken myoblasts derived from embryonic tissue. Myoblasts are derived from satellite cells, which are a precursor to myogenesis [8]. In the quiescent satellite cell stage, PAX7, a critical marker of undifferentiated myoblasts, is highly expressed. After activation, myoblasts start to proliferate, decrease expression of PAX7, and increase expression of MYOD, a myogenic regulatory factor (MRF). Then they enter the stage of terminal differentiation. In this stage, expression of MYOD decreases, and expression of markers of terminal differentiation such as Myogenin and Desmin increases. Eventually myoblast cells form new myotubes, which in turn form new myofibers [9]. Therefore, myoblasts are closely related to muscle growth, which is an economically important trait of domestic animals.

MiR-743a-5p regulates differentiation of myoblast by targeting Mob1b in skeletal muscle development and regeneration

“…MiRNAs which are negative in proliferation usually display a positive role in myoblast differentiation. 34 MiR-743a-5p actually enhanced differentiation and promoted myotubes formation. It displayed a potential positive role in differentiation.…”

Identification of microRNA-Associated-ceRNA Networks Regulating Crop Milk Production in Pigeon (Columba livia)

“…We noticed that many miRNAs involved in cell proliferation and migration have been identified, such as miR-20b-5p, which was proved to participate in the epithelial mesenchymal transition (EMT), migration and invasion process of prostate cancer [ 45 ]. MiR-146b-5p played a regulatory role in the proliferation and differentiation of human fibroblasts and chicken myoblasts [ 46 , 47 ]. MiR-21-5p is up-regulated in a variety of cancer cells such as non-small cell lung cancer, Hepatocellular carcinoma, melanoma, and colon adenocarcinoma cells, and has been shown to promote cell proliferation [ 48 , 49 , 50 , 51 ].…”

Regulation of Non-Coding RNA in the Growth and Development of Skeletal Muscle in Domestic Chickens

“…During the embryonic period, myoblasts grow and transform into different types of muscle fibers, which determine the number of muscle fibers after birth [ 57 ]. Many studies have shown that miRNAs affect the growth and differentiation of myoblasts in vitro.…”