Discovery of MicroRNAs Associated with Myogenesis by Deep Sequencing of Serial Developmental Skeletal Muscles in Pigs

Abstract

MicroRNAs (miRNAs) are short, single-stranded non-coding RNAs that repress their target genes by binding their 3′ UTRs. These RNAs play critical roles in myogenesis. To gain knowledge about miRNAs involved in the regulation of myogenesis, porcine longissimus muscles were collected from 18 developmental stages (33-, 40-, 45-, 50-, 55-, 60-, 65-, 70-, 75-, 80-, 85-, 90-, 95-, 100- and 105-day post-gestation fetuses, 0 and 10-day postnatal piglets and adult pigs) to identify miRNAs using Solexa sequencing technology. We detected 197 known miRNAs and 78 novel miRNAs according to comparison with known miRNAs in the miRBase (release 17.0) database. Moreover, variations in sequence length and single nucleotide polymorphisms were also observed in 110 known miRNAs. Expression analysis of the 11 most abundant miRNAs were conducted using quantitative PCR (qPCR) in eleven tissues (longissimus muscles, leg muscles, heart, liver, spleen, lung, kidney, stomach, small intestine and colon), and the results revealed that ssc-miR-378, ssc-miR-1 and ssc-miR-206 were abundantly expressed in skeletal muscles. During skeletal muscle development, the expression level of ssc-miR-378 was low at 33 days post-coitus (dpc), increased at 65 and 90 dpc, peaked at postnatal day 0, and finally declined and maintained a comparatively stable level. This expression profile suggested that ssc-miR-378 was a new candidate miRNA for myogenesis and participated in skeletal muscle development in pigs. Target prediction and KEGG pathway analysis suggested that bone morphogenetic protein 2 (BMP2) and mitogen-activated protein kinase 1 (MAPK1), both of which were relevant to proliferation and differentiation, might be the potential targets of miR-378. Luciferase activities of report vectors containing the 3′UTR of porcine BMP2 or MAPK1 were downregulated by miR-378, which suggested that miR-378 probably regulated myogenesis though the regulation of these two genes.

The long and short of non-coding RNAs during post-natal growth and differentiation of skeletal muscles: Focus on lncRNA and miRNAs

“…The associated in vivo changes in vascular development, neuromuscular maturation, as well as complex modulation of the extracellular matrix during this rapid growth and maturation of skeletal muscle tissue, also needs to be considered. Greater insight into interactions between the many forms of RNAs and proteins that regulate muscle growth and maintain homeostasis, is of central importance to livestock industries related to improvement of meat quality and production using molecular approaches (Hou et al, 2012;Li et al, 2012) and to clinical muscle disorders. For example, many ncRNAs have been identified in cases of aberrant muscle growth and muscle atrophy, including genetic diseases as Duchenne muscular dystrophy (Neguembor et al, 2014;Twayana et al, 2013), inflammatory conditions like cachexia (Soares et al, 2014) and age-related muscle wasting (Drummond et al, 2011); therapeutic modulation of many ncRNAs by exercise has recently been demonstrated (Zacharewicz et al, 2014).…”

Arsenic inhibits hedgehog signaling during P19 cell differentiation

“…Gli2 expression can also be decreased by oxidative stress in both animal models and in cultured astrocytes (Xia et al , 2012; Ji et al , 2012). Studies have shown that microRNAs participate in development and cell differentiation through regulating growth related gene expression (Kim, 2005; Hou et al , 2012; Kawahara et al , 2012). Moreover, a recent study shows that miR-326 represses Shh signaling activity by directly targeting Gli2 expression in embryonic lung epithelium cells (Jiang et al , 2014).…”

MiR-27b Promotes Muscle Development by Inhibiting MDFI Expression

“…Proliferation and differentiation are mutually exclusive during muscle regeneration, and miRNAs are critically involved in balancing these two processes [43]. Many miRNAs, such as miR-1, miR-133, miR-29, miR-214, miR-206, miR-486, miR-208b, and miR-499 were involved in the regulation of skeletal myogenesis by binding to its target genes [44, 45]. For example, in mice, miR-1 and miR-133 are clustered on the same chromosomal loci and transcribed together in a tissue-specific manner during development, but miR-133 enhances proliferation by repressing serum response factor, whereas miR-1 promotes myogenesis through repressing histone deacetylase 4 [17].…”