<i>NCAPG</i> Regulates Myogenesis in Sheep, and SNPs Located in Its Putative Promoter Region Are Associated with Growth and Development Traits
Previously, <i>NCAPG</i> was identified as a candidate gene associated with sheep growth traits. This study aimed to investigate the direct role of <i>NCAPG</i> in regulating myogenesis in embryonic myoblast cells and to investigate the association between single-nucleotide p...
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MDPI AG
2023-10-01
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| Online Access: | https://www.mdpi.com/2076-2615/13/20/3173 |
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| author | Zehu Yuan Ling Ge Pengwei Su Yifei Gu Weihao Chen Xiukai Cao Shanhe Wang Xiaoyang Lv Tesfaye Getachew Joram M. Mwacharo Aynalem Haile Wei Sun |
| author_facet | Zehu Yuan Ling Ge Pengwei Su Yifei Gu Weihao Chen Xiukai Cao Shanhe Wang Xiaoyang Lv Tesfaye Getachew Joram M. Mwacharo Aynalem Haile Wei Sun |
| author_sort | Zehu Yuan |
| collection | DOAJ |
| description | Previously, <i>NCAPG</i> was identified as a candidate gene associated with sheep growth traits. This study aimed to investigate the direct role of <i>NCAPG</i> in regulating myogenesis in embryonic myoblast cells and to investigate the association between single-nucleotide polymorphisms (SNPs) in its promoter region and sheep growth traits. The function of <i>NCAPG</i> in myoblast proliferation and differentiation was detected after small interfering RNAs (siRNAs) knocked down the expression of <i>NCAPG</i>. Cell proliferation was detected using CCK-8 assay, EdU proliferation assay, and flow cytometry cell cycle analysis. Cell differentiation was detected via cell immunofluorescence and the quantification of myogenic regulatory factors (MRFs). SNPs in the promoter region were detected using Sanger sequencing and genotyped using the improved multiplex ligation detection reaction (iMLDR<sup>®</sup>) technique. As a result, a notable decrease (<i>p</i> < 0.01) in the percentage of EdU-positive cells in the siRNA-694-treated group was observed. A significant decrease (<i>p</i> < 0.01) in cell viability after treatment with siRNA-694 for 48 h and 72 h was detected using the CCK-8 method. The quantity of S-phase cells in the siRNA-694 treatment group was significantly decreased (<i>p</i> < 0.01). After interfering with <i>NCAPG</i> in myoblasts during induced differentiation, the relative expression levels of MRFs were markedly (<i>p</i> < 0.05 or <i>p</i> < 0.01) reduced compared with the control group on days 5–7. The myoblast differentiation in the siRNA-694 treatment group was obviously suppressed compared with the control group. SNP1, SNP2, SNP3, and SNP4 were significantly (<i>p</i> < 0.05) associated with all traits except body weight measured at birth and one month of age. SNP5 was significantly (<i>p</i> < 0.05) associated with body weight, body height, and body length in six-month-old sheep. In conclusion, interfering with <i>NCAPG</i> can inhibit the proliferation and differentiation of ovine embryonic myoblasts. SNPs in its promoter region can serve as potential useful markers for selecting sheep growth traits. |
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| last_indexed | 2024-03-10T21:30:28Z |
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| spelling | doaj.art-87e5685a6d754f728cfb799ade2dc8fc2023-11-19T15:24:11ZengMDPI AGAnimals2076-26152023-10-011320317310.3390/ani13203173<i>NCAPG</i> Regulates Myogenesis in Sheep, and SNPs Located in Its Putative Promoter Region Are Associated with Growth and Development TraitsZehu Yuan0Ling Ge1Pengwei Su2Yifei Gu3Weihao Chen4Xiukai Cao5Shanhe Wang6Xiaoyang Lv7Tesfaye Getachew8Joram M. Mwacharo9Aynalem Haile10Wei Sun11Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, ChinaJoint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, ChinaJoint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, ChinaJoint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, ChinaJoint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, ChinaJoint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, ChinaJoint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, ChinaJoint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, ChinaInternational Centre for Agricultural Research in the Dry Areas, Addis Ababa 999047, EthiopiaInternational Centre for Agricultural Research in the Dry Areas, Addis Ababa 999047, EthiopiaInternational Centre for Agricultural Research in the Dry Areas, Addis Ababa 999047, EthiopiaJoint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, ChinaPreviously, <i>NCAPG</i> was identified as a candidate gene associated with sheep growth traits. This study aimed to investigate the direct role of <i>NCAPG</i> in regulating myogenesis in embryonic myoblast cells and to investigate the association between single-nucleotide polymorphisms (SNPs) in its promoter region and sheep growth traits. The function of <i>NCAPG</i> in myoblast proliferation and differentiation was detected after small interfering RNAs (siRNAs) knocked down the expression of <i>NCAPG</i>. Cell proliferation was detected using CCK-8 assay, EdU proliferation assay, and flow cytometry cell cycle analysis. Cell differentiation was detected via cell immunofluorescence and the quantification of myogenic regulatory factors (MRFs). SNPs in the promoter region were detected using Sanger sequencing and genotyped using the improved multiplex ligation detection reaction (iMLDR<sup>®</sup>) technique. As a result, a notable decrease (<i>p</i> < 0.01) in the percentage of EdU-positive cells in the siRNA-694-treated group was observed. A significant decrease (<i>p</i> < 0.01) in cell viability after treatment with siRNA-694 for 48 h and 72 h was detected using the CCK-8 method. The quantity of S-phase cells in the siRNA-694 treatment group was significantly decreased (<i>p</i> < 0.01). After interfering with <i>NCAPG</i> in myoblasts during induced differentiation, the relative expression levels of MRFs were markedly (<i>p</i> < 0.05 or <i>p</i> < 0.01) reduced compared with the control group on days 5–7. The myoblast differentiation in the siRNA-694 treatment group was obviously suppressed compared with the control group. SNP1, SNP2, SNP3, and SNP4 were significantly (<i>p</i> < 0.05) associated with all traits except body weight measured at birth and one month of age. SNP5 was significantly (<i>p</i> < 0.05) associated with body weight, body height, and body length in six-month-old sheep. In conclusion, interfering with <i>NCAPG</i> can inhibit the proliferation and differentiation of ovine embryonic myoblasts. SNPs in its promoter region can serve as potential useful markers for selecting sheep growth traits.https://www.mdpi.com/2076-2615/13/20/3173sheep growth and developmentmyoblastsNCAPGSNP |
| spellingShingle | Zehu Yuan Ling Ge Pengwei Su Yifei Gu Weihao Chen Xiukai Cao Shanhe Wang Xiaoyang Lv Tesfaye Getachew Joram M. Mwacharo Aynalem Haile Wei Sun <i>NCAPG</i> Regulates Myogenesis in Sheep, and SNPs Located in Its Putative Promoter Region Are Associated with Growth and Development Traits Animals sheep growth and development myoblasts NCAPG SNP |
| title | <i>NCAPG</i> Regulates Myogenesis in Sheep, and SNPs Located in Its Putative Promoter Region Are Associated with Growth and Development Traits |
| title_full | <i>NCAPG</i> Regulates Myogenesis in Sheep, and SNPs Located in Its Putative Promoter Region Are Associated with Growth and Development Traits |
| title_fullStr | <i>NCAPG</i> Regulates Myogenesis in Sheep, and SNPs Located in Its Putative Promoter Region Are Associated with Growth and Development Traits |
| title_full_unstemmed | <i>NCAPG</i> Regulates Myogenesis in Sheep, and SNPs Located in Its Putative Promoter Region Are Associated with Growth and Development Traits |
| title_short | <i>NCAPG</i> Regulates Myogenesis in Sheep, and SNPs Located in Its Putative Promoter Region Are Associated with Growth and Development Traits |
| title_sort | i ncapg i regulates myogenesis in sheep and snps located in its putative promoter region are associated with growth and development traits |
| topic | sheep growth and development myoblasts NCAPG SNP |
| url | https://www.mdpi.com/2076-2615/13/20/3173 |
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