Breed-dependent microRNA expression in the primary culture of skeletal muscle cells subjected to myogenic differentiation

Abstract

BackgroundSkeletal muscle in livestock develops into meat, an important source of protein and other nutrients for human consumption. The muscle is largely composed of a fixed number of multinucleated myofibers determined during late gestation and remains constant postnatally. A population of postnatal muscle stem cells, called satellite cells, gives rise to myoblast cells that can fuse with the existing myofibers, thus increasing their size. This requires a delicate balance of transcription and growth factors and specific microRNA (miRNA) expressed by satellite cells and their supporting cells from the muscle stem cell niche. The role of transcription and growth factors in bovine myogenesis is well-characterized; however, very little is known about the miRNA activity during this process. We have hypothesized that the expression of miRNA can vary between primary cultures of skeletal muscle cells isolated from the semitendinosus muscles of different cattle breeds and subjected to myogenic differentiation.ResultsAfter a 6-day myogenic differentiation of cells isolated from the muscles of the examined cattle breeds, we found statistically significant differences in the number of myotubes between Hereford (HER)/Limousine (LIM) beef breeds and the Holstein-Friesian (HF) dairy breed (p ≤ 0.001). The microarray analysis revealed differences in the expression of 23 miRNA among the aforementioned primary cultures. On the basis of a functional analysis, we assigned 9 miRNA as molecules responsible for differentiation progression (miR-1, -128a, -133a, -133b, -139, -206, -222, -486, and -503). The target gene prediction and functional analysis revealed 59 miRNA-related genes belonging to the muscle organ development process.ConclusionThe number of myotubes and the miRNA expression in the primary cultures of skeletal muscle cells derived from the semitendinosus muscles of the HER/LIM beef cattle breeds and the HF dairy breed vary when cells are subjected to myogenic differentiation. The net effect of the identified miRNA and their target gene action should be considered the result of the breed-dependent activity of satellite cells and muscle stem cell niche cells and their mutual interactions, which putatively can be engaged in the formation of a larger number of myotubes in beef cattle-related cells (HER/LIM) during in vitro myogenesis.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-4492-5) contains supplementary material, which is available to authorized users.

Transcriptomic Profile of Primary Culture of Skeletal Muscle Cells Isolated from Semitendinosus Muscle of Beef and Dairy Bulls

“…In the case of the culture of LIM and HER breeds, myotubes formed as a result of myoblasts fusion were perceptible already from the fourth day of differentiation, whereas the myotube formation in the HF breed can be only observed on the sixth day of differentiation ( Figure S2 ). Previously published research performed on a similar primary skeletal muscle cell culture derived from the same individuals showed statistically significantly higher numbers of myotubes in beef breeds in comparison to the dairy breed [ 16 ].…”

Transcriptomic profile of semitendinosus muscle of bulls of different breed and performance

“…The experimental group consisted of bulls belonging to three cattle breeds differing in meat production and performance: LIM (beef breed, capable of high-meat low-fat production; n = 4), HER (beef breed, capable of high-meat high-fat production; n = 4), and HF (dairy breed; n = 4) used as a reference. The bulls were housed, fed, and slaughtered at the age of 15 months as described earlier (Sadkowski et al 2009a , 2018 ). The muscle tissue samples were compared individually—4 biological repetitions.…”

Tissue Engineering for Clean Meat Production

“…miRNA, short RNA molecules that inhibit genes with complementary mRNA, play a crucial role in the regulation of muscle tissue development, including myogenesis (Wang et al, 2015;Dai et al, 2016;Zhang et al, 2017;Sadkowski et al, 2018) and adipogenesis (Yan et al, 2013). Several naturally occurring miRNA molecules have been shown to regulate myogenesis and adipogenesis in farm animals, including regulation of PPARG and C/EBPα expression (Romao et al, 2011) and lipid metabolism (Wilfred et al, 2007).…”