Abstract
MicroRNAs (miRNAs) play a vital role in muscle development by binding to messenger RNAs (mRNAs). Based on prenatal skeletal muscle at 33, 65 and 90 days post-coitus (dpc) from Landrace, Tongcheng and Wuzhishan pigs, we carried out integrated analysis of miRNA and mRNA expression profiling. We identified 33, 18 and 67 differentially expressed miRNAs and 290, 91 and 502 mRNA targets in Landrace, Tongcheng and Wuzhishan pigs, respectively. Subsequently, 12 mRNAs and 3 miRNAs differentially expressed were validated using quantitative real-time PCR (qPCR), and 5 predicted miRNA targets were confirmed via dual luciferase reporter or western blot assays. We identified a set of miRNAs and mRNA genes differentially expressed in muscle development. Gene ontology (GO) enrichment analysis suggests that the miRNA targets are primarily involved in muscle contraction, muscle development and negative regulation of cell proliferation. Our data indicated that more mRNAs are regulated by miRNAs at earlier stages than at later stages of muscle development. Landrace and Tongcheng pigs also had longer phases of myoblast proliferation than Wuzhishan pigs. This study will be helpful to further explore miRNA-mRNA interactions in myogenesis and aid to uncover the molecular mechanisms of muscle development and phenotype variance in pigs.
Highlights
MicroRNAs are evolutionary conserved, small (~22 nt) non-coding RNAs that are found in all metazoans and regulate diverse biological processes1–5. miRNAs regulate gene expression by base pairing with their target messenger RNAs (mRNAs), leading to mRNA cleavage or translational repression[5,6]
Postnatal muscle growth is determined by the total number of fibers (TNF), which is fixed before birth
In Landrace pigs and Tongcheng pigs, miR-206 peaked at 90 dpc, but it peaked at 65 dpc in Wuzhishan pigs
Summary
MicroRNAs (miRNAs) are evolutionary conserved, small (~22 nt) non-coding RNAs that are found in all metazoans and regulate diverse biological processes1–5. miRNAs regulate gene expression by base pairing with their target mRNAs, leading to mRNA cleavage or translational repression[5,6]. Several computational methods (PicTar, Target-Scan and miRanda) have been developed for the prediction of miRNA targets[6,7,8,9] These programs are valuable for guiding laboratory experiments, they lack sensitivity and specificity[10,11]. The miRNAs represent a newly recognized level of regulation of gene expression mediating skeletal muscle development. The function of miRNAs in skeletal muscle development and phenotypic variance has remained largely unclear. Prenatal skeletal muscle development (myogenesis) is an ideal paradigm for understanding the molecular basis of cell lineage establishment and differentiation into specialized structures[40,41]. In the pig (Sus scrofa), two waves of muscle fiber formation occur over relatively long periods of time compared to other laboratory animals, making the pig a good animal model for studying myogenesis[45]. Wuzhishan pigs, which are a Chinese miniature breed, have been recognized as an attractive experimental animal for a wide range of research fields (adults weigh < 40 kg)[54,55]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
