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Wnt antagonist, secreted frizzled-related protein 1, is involved in prenatal skeletal muscle development and is a target of miRNA-1/206 in pigs.

Yang Y, Sun W, Wang R, Lei C, Zhou R, Tang Z, Li K - BMC Mol. Biol. (2015)

Bottom Line: The expression level of the SFRP1 was significantly higher in the embryonic skeletal compared with postnatal skeletal muscle, whereas miR-206 showed the inverse pattern of expression.Dual luciferase assay and Western-blot results demonstrated that SFRP1 was a target of miR-1/206 in porcine iliac endothelial cells.Our results indicate that the SFRP1 gene is regulated by miR-1/206 and potentially affects skeletal muscle development.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Farm Animal Genetic Resources and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100193, P.R. China. yangyalan1988@126.com.

ABSTRACT

Background: The Wnt signaling pathway is involved in the control of cell proliferation and differentiation during skeletal muscle development. Secreted frizzled-related proteins (SFRPs), such as SFRP1, function as inhibitors of Wnt signaling. MicroRNA-1/206(miRNA-1/206) is specifically expressed in skeletal muscle and play a critical role in myogenesis. The miRNA-mRNA profiles and bioinformatics study suggested that the SFRP1 gene was potentially regulated by miRNA-1/206 during porcine skeletal muscle development.

Methods: To understand the function of SFRP1 and miRNA-1/206 in swine myogenesis, we first predicted the targets of miRNA-1/206 with the TargetScan and PicTar programs, and analyzed the molecular characterization of the porcine SFRP1 gene. We performed a temporal-spatial expression analysis of SFRP1 mRNA and miRNA-206 in Tongcheng pigs (a Chinese indigenous breed) by quantitative real-time polymerase chain reaction, and conducted the co-expression analyses of SFRP1 and miRNA-1/206. Subsequently, the interaction between SFRP1 and miRNA-1/206 was validated via dual luciferase and Western blot assays.

Results: The bioinformatics analysis predicted SFRP1 to be a target of miRNA-1/206. The expression level of the SFRP1 was highly varied across numerous pig tissues and it was down-regulated during porcine skeletal muscle development. The expression level of the SFRP1 was significantly higher in the embryonic skeletal compared with postnatal skeletal muscle, whereas miR-206 showed the inverse pattern of expression. A significant negative correlation was observed between the expression of miR-1/206 and SFRP1 during porcine skeletal muscle development (p <0.05). Dual luciferase assay and Western-blot results demonstrated that SFRP1 was a target of miR-1/206 in porcine iliac endothelial cells.

Conclusions: Our results indicate that the SFRP1 gene is regulated by miR-1/206 and potentially affects skeletal muscle development. These findings increase understanding of the biological functions and the regulation of the SFRP1 gene in mammals.

Show MeSH
Top 10 GO biological process terms significantly enriched in for target genes of miRNA-1/206. GO analysis was conducted with a DAVID functional annotation program.
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Fig1: Top 10 GO biological process terms significantly enriched in for target genes of miRNA-1/206. GO analysis was conducted with a DAVID functional annotation program.

Mentions: The candidate targets of miRNA-1/206 were predicted using TargetScan and PicTar programs. 258 candidate targets were predicted by both programs. To explore the biological function of the candidate genes, a DAVID functional annotation analysis was performed using the thresholds EASE adjusted to p < 0.05. The Gene Ontology (GO) analysis revealed that these targets were significantly enriched in the regulation of transcription, positive regulation of macromolecule metabolic processes, positive regulation of nitrogen compound metabolic processes, positive regulation of cellular biosynthetic processes and other biological processes (Figure 1). KEGG pathway analysis showed that these targets were significantly enriched in four pathways (p <0.05), which were SNARE interactions in vesicular transport, the Wnt signaling pathway, small cell lung cancer and the Neurotrophin signaling pathway (Table 2). Seven putative targets, including CCND1, SFRP1, CCND2, PPP2R5A, NFAT5, DAAM1 and FZD7, participated in the Wnt signaling pathway. Therefore, the SFRP1 gene, which was predicted to be a target of miRNA-1/206 and was involved in the Wnt signaling pathway, was selected for further study.Figure 1


Wnt antagonist, secreted frizzled-related protein 1, is involved in prenatal skeletal muscle development and is a target of miRNA-1/206 in pigs.

Yang Y, Sun W, Wang R, Lei C, Zhou R, Tang Z, Li K - BMC Mol. Biol. (2015)

Top 10 GO biological process terms significantly enriched in for target genes of miRNA-1/206. GO analysis was conducted with a DAVID functional annotation program.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4359577&req=5

Fig1: Top 10 GO biological process terms significantly enriched in for target genes of miRNA-1/206. GO analysis was conducted with a DAVID functional annotation program.
Mentions: The candidate targets of miRNA-1/206 were predicted using TargetScan and PicTar programs. 258 candidate targets were predicted by both programs. To explore the biological function of the candidate genes, a DAVID functional annotation analysis was performed using the thresholds EASE adjusted to p < 0.05. The Gene Ontology (GO) analysis revealed that these targets were significantly enriched in the regulation of transcription, positive regulation of macromolecule metabolic processes, positive regulation of nitrogen compound metabolic processes, positive regulation of cellular biosynthetic processes and other biological processes (Figure 1). KEGG pathway analysis showed that these targets were significantly enriched in four pathways (p <0.05), which were SNARE interactions in vesicular transport, the Wnt signaling pathway, small cell lung cancer and the Neurotrophin signaling pathway (Table 2). Seven putative targets, including CCND1, SFRP1, CCND2, PPP2R5A, NFAT5, DAAM1 and FZD7, participated in the Wnt signaling pathway. Therefore, the SFRP1 gene, which was predicted to be a target of miRNA-1/206 and was involved in the Wnt signaling pathway, was selected for further study.Figure 1

Bottom Line: The expression level of the SFRP1 was significantly higher in the embryonic skeletal compared with postnatal skeletal muscle, whereas miR-206 showed the inverse pattern of expression.Dual luciferase assay and Western-blot results demonstrated that SFRP1 was a target of miR-1/206 in porcine iliac endothelial cells.Our results indicate that the SFRP1 gene is regulated by miR-1/206 and potentially affects skeletal muscle development.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Farm Animal Genetic Resources and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100193, P.R. China. yangyalan1988@126.com.

ABSTRACT

Background: The Wnt signaling pathway is involved in the control of cell proliferation and differentiation during skeletal muscle development. Secreted frizzled-related proteins (SFRPs), such as SFRP1, function as inhibitors of Wnt signaling. MicroRNA-1/206(miRNA-1/206) is specifically expressed in skeletal muscle and play a critical role in myogenesis. The miRNA-mRNA profiles and bioinformatics study suggested that the SFRP1 gene was potentially regulated by miRNA-1/206 during porcine skeletal muscle development.

Methods: To understand the function of SFRP1 and miRNA-1/206 in swine myogenesis, we first predicted the targets of miRNA-1/206 with the TargetScan and PicTar programs, and analyzed the molecular characterization of the porcine SFRP1 gene. We performed a temporal-spatial expression analysis of SFRP1 mRNA and miRNA-206 in Tongcheng pigs (a Chinese indigenous breed) by quantitative real-time polymerase chain reaction, and conducted the co-expression analyses of SFRP1 and miRNA-1/206. Subsequently, the interaction between SFRP1 and miRNA-1/206 was validated via dual luciferase and Western blot assays.

Results: The bioinformatics analysis predicted SFRP1 to be a target of miRNA-1/206. The expression level of the SFRP1 was highly varied across numerous pig tissues and it was down-regulated during porcine skeletal muscle development. The expression level of the SFRP1 was significantly higher in the embryonic skeletal compared with postnatal skeletal muscle, whereas miR-206 showed the inverse pattern of expression. A significant negative correlation was observed between the expression of miR-1/206 and SFRP1 during porcine skeletal muscle development (p <0.05). Dual luciferase assay and Western-blot results demonstrated that SFRP1 was a target of miR-1/206 in porcine iliac endothelial cells.

Conclusions: Our results indicate that the SFRP1 gene is regulated by miR-1/206 and potentially affects skeletal muscle development. These findings increase understanding of the biological functions and the regulation of the SFRP1 gene in mammals.

Show MeSH