Loss of WTAP Impairs Early Parthenogenetic Embryo Development
m<sup>6</sup>A is one of the most common and abundant modifications of RNA molecules present in eukaryotes. The methyltransferase complex, consisting of methyltransferase-like 3 (METTL3), METTL14, and WTAP, is responsible for the m<sup>6</sup>A modification of RNA. WTAP was i...
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2021-06-01
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| author | Jindong Hao Siyi Huang Dongxu Wang Yongxun Jin Mingjun Zhang Jiabao Zhang Xianfeng Yu |
| author_facet | Jindong Hao Siyi Huang Dongxu Wang Yongxun Jin Mingjun Zhang Jiabao Zhang Xianfeng Yu |
| author_sort | Jindong Hao |
| collection | DOAJ |
| description | m<sup>6</sup>A is one of the most common and abundant modifications of RNA molecules present in eukaryotes. The methyltransferase complex, consisting of methyltransferase-like 3 (METTL3), METTL14, and WTAP, is responsible for the m<sup>6</sup>A modification of RNA. WTAP was identified as an mRNA splicing regulator. Its role as a regulatory subunit of the m<sup>6</sup>A methyltransferase complex in embryonic development remains largely unknown. To investigate the role of WTAP in porcine early embryonic development, si-WTAP was microinjected into porcine parthenogenetic zygotes. WTAP knockdown significantly reduced the blastocyst rate and global m<sup>6</sup>A levels, but did not affect the cleavage rate. Betaine was supplemented into the in vitro culture (IVC) to increase the m<sup>6</sup>A levels. Betaine significantly increased the global m<sup>6</sup>A levels but did not affect the blastocyst rate. Furthermore, the pluripotency genes, including <i>OCT4</i>, <i>SOX2</i>, and <i>NANOG</i>, were downregulated following WTAP knockdown. The apoptotic genes <i>BAX</i> and <i>CASPASE 3</i> were upregulated, while the anti-apoptotic gene <i>BCL2</i> was downregulated in WTAP knockdown blastocysts. TUNEL staining revealed that the number of apoptotic cells was significantly increased following WTAP knockdown. Our study indicated that WTAP has an indispensable role in porcine early embryonic development. |
| first_indexed | 2024-03-10T10:42:58Z |
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| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-10T10:42:58Z |
| publishDate | 2021-06-01 |
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| series | Animals |
| spelling | doaj.art-6ea91e1ebfe44d3ebc12ec62cef839c12023-11-21T22:46:54ZengMDPI AGAnimals2076-26152021-06-01116167510.3390/ani11061675Loss of WTAP Impairs Early Parthenogenetic Embryo DevelopmentJindong Hao0Siyi Huang1Dongxu Wang2Yongxun Jin3Mingjun Zhang4Jiabao Zhang5Xianfeng Yu6Jilin Provincial Key Laboratory of Animal Model, College of Animal Science, Jilin University, Changchun 130062, ChinaJilin Provincial Key Laboratory of Animal Model, College of Animal Science, Jilin University, Changchun 130062, ChinaJilin Provincial Key Laboratory of Animal Model, College of Animal Science, Jilin University, Changchun 130062, ChinaJilin Provincial Key Laboratory of Animal Model, College of Animal Science, Jilin University, Changchun 130062, ChinaJilin Provincial Key Laboratory of Animal Model, College of Animal Science, Jilin University, Changchun 130062, ChinaJilin Provincial Key Laboratory of Animal Model, College of Animal Science, Jilin University, Changchun 130062, ChinaJilin Provincial Key Laboratory of Animal Model, College of Animal Science, Jilin University, Changchun 130062, Chinam<sup>6</sup>A is one of the most common and abundant modifications of RNA molecules present in eukaryotes. The methyltransferase complex, consisting of methyltransferase-like 3 (METTL3), METTL14, and WTAP, is responsible for the m<sup>6</sup>A modification of RNA. WTAP was identified as an mRNA splicing regulator. Its role as a regulatory subunit of the m<sup>6</sup>A methyltransferase complex in embryonic development remains largely unknown. To investigate the role of WTAP in porcine early embryonic development, si-WTAP was microinjected into porcine parthenogenetic zygotes. WTAP knockdown significantly reduced the blastocyst rate and global m<sup>6</sup>A levels, but did not affect the cleavage rate. Betaine was supplemented into the in vitro culture (IVC) to increase the m<sup>6</sup>A levels. Betaine significantly increased the global m<sup>6</sup>A levels but did not affect the blastocyst rate. Furthermore, the pluripotency genes, including <i>OCT4</i>, <i>SOX2</i>, and <i>NANOG</i>, were downregulated following WTAP knockdown. The apoptotic genes <i>BAX</i> and <i>CASPASE 3</i> were upregulated, while the anti-apoptotic gene <i>BCL2</i> was downregulated in WTAP knockdown blastocysts. TUNEL staining revealed that the number of apoptotic cells was significantly increased following WTAP knockdown. Our study indicated that WTAP has an indispensable role in porcine early embryonic development.https://www.mdpi.com/2076-2615/11/6/1675m<sup>6</sup>AWTAPporcineembryo developmentparthenogenetic |
| spellingShingle | Jindong Hao Siyi Huang Dongxu Wang Yongxun Jin Mingjun Zhang Jiabao Zhang Xianfeng Yu Loss of WTAP Impairs Early Parthenogenetic Embryo Development Animals m<sup>6</sup>A WTAP porcine embryo development parthenogenetic |
| title | Loss of WTAP Impairs Early Parthenogenetic Embryo Development |
| title_full | Loss of WTAP Impairs Early Parthenogenetic Embryo Development |
| title_fullStr | Loss of WTAP Impairs Early Parthenogenetic Embryo Development |
| title_full_unstemmed | Loss of WTAP Impairs Early Parthenogenetic Embryo Development |
| title_short | Loss of WTAP Impairs Early Parthenogenetic Embryo Development |
| title_sort | loss of wtap impairs early parthenogenetic embryo development |
| topic | m<sup>6</sup>A WTAP porcine embryo development parthenogenetic |
| url | https://www.mdpi.com/2076-2615/11/6/1675 |
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