Production of mouse offspring from inactivated spermatozoa using horse PLCζ mRNA
Improving artificial oocyte activation is essential for assisted reproduction or animal biotechnology that can obtain healthy offspring with a high success rate. Here, we examined whether intracytoplasmic injection of equine sperm-specific phospholipase C zeta (ePLCζ) mRNA, the PLCζ with the stronge...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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The Society for Reproduction and Development
2019-12-01
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| Series: | The Journal of Reproduction and Development |
| Subjects: | |
| Online Access: | https://www.jstage.jst.go.jp/article/jrd/66/1/66_2019-043/_pdf/-char/en |
| _version_ | 1797615262369841152 |
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| author | Yunosuke YAMAMOTO Naoki HIROSE Satoshi KAMIMURA Sayaka WAKAYAMA Junya ITO Masatoshi OOGA Teruhiko WAKAYAMA |
| author_facet | Yunosuke YAMAMOTO Naoki HIROSE Satoshi KAMIMURA Sayaka WAKAYAMA Junya ITO Masatoshi OOGA Teruhiko WAKAYAMA |
| author_sort | Yunosuke YAMAMOTO |
| collection | DOAJ |
| description | Improving artificial oocyte activation is essential for assisted reproduction or animal biotechnology that can obtain healthy offspring with a high success rate. Here, we examined whether intracytoplasmic injection of equine sperm-specific phospholipase C zeta (ePLCζ) mRNA, the PLCζ with the strongest oocyte activation potential in mammals, could improve the mouse oocyte activation rate and subsequent embryonic development using inactivated spermatozoa. mRNA of mouse PLCζ (mPLCζ) or ePLCζ were injected into mouse oocytes to determine the optimal mRNA concentration to maximize the oocyte activation rate and developmental rate of parthenogenetic embryos in vitro. Full-term development was examined using NaOH-treated inactive spermatozoa using the optimal activation method. We found that the most optimal ePLCζ mRNA concentration was 0.1 ng/µl for mouse oocyte activation, which was ten times stronger than mPLCζ mRNA. The concentration did not affect parthenogenetic embryo development in vitro. Relatively normal blastocysts were obtained with the same developmental rate (52–53% or 48–51%, respectively) when inactive spermatozoa were injected into activated oocytes using ePLCζ or mPLCζ mRNA injection. However, the birth rate after embryo transfer was slightly but significantly decreased in oocytes activated by ePLCζ mRNA (24%) compared to mPLCζ mRNA (37%) or strontium treatment (40%) activation. These results suggest that the higher activation rate does not always correlate the higher birth rate, and some mechanisms might exist in the oocyte activation process that could affect the later developmental stages like full-term development. |
| first_indexed | 2024-03-11T07:23:48Z |
| format | Article |
| id | doaj.art-ab27148701a24c6dbe53d10acd235fed |
| institution | Directory Open Access Journal |
| issn | 0916-8818 1348-4400 |
| language | English |
| last_indexed | 2024-03-11T07:23:48Z |
| publishDate | 2019-12-01 |
| publisher | The Society for Reproduction and Development |
| record_format | Article |
| series | The Journal of Reproduction and Development |
| spelling | doaj.art-ab27148701a24c6dbe53d10acd235fed2023-11-17T07:44:30ZengThe Society for Reproduction and DevelopmentThe Journal of Reproduction and Development0916-88181348-44002019-12-01661677310.1262/jrd.2019-043jrdProduction of mouse offspring from inactivated spermatozoa using horse PLCζ mRNAYunosuke YAMAMOTO0Naoki HIROSE1Satoshi KAMIMURA2Sayaka WAKAYAMA3Junya ITO4Masatoshi OOGA5Teruhiko WAKAYAMA6Faculty of Life and Environmental Science, University of Yamanashi, Yamanashi 400-8510, JapanFaculty of Life and Environmental Science, University of Yamanashi, Yamanashi 400-8510, JapanFaculty of Life and Environmental Science, University of Yamanashi, Yamanashi 400-8510, JapanAdvanced Biotechnology Center, University of Yamanashi, Yamanashi 400-8510, JapanLaboratory of Animal Reproduction, Graduate School of Veterinary Science, Azabu University, Kanagawa 252-5201, JapanFaculty of Life and Environmental Science, University of Yamanashi, Yamanashi 400-8510, JapanFaculty of Life and Environmental Science, University of Yamanashi, Yamanashi 400-8510, JapanImproving artificial oocyte activation is essential for assisted reproduction or animal biotechnology that can obtain healthy offspring with a high success rate. Here, we examined whether intracytoplasmic injection of equine sperm-specific phospholipase C zeta (ePLCζ) mRNA, the PLCζ with the strongest oocyte activation potential in mammals, could improve the mouse oocyte activation rate and subsequent embryonic development using inactivated spermatozoa. mRNA of mouse PLCζ (mPLCζ) or ePLCζ were injected into mouse oocytes to determine the optimal mRNA concentration to maximize the oocyte activation rate and developmental rate of parthenogenetic embryos in vitro. Full-term development was examined using NaOH-treated inactive spermatozoa using the optimal activation method. We found that the most optimal ePLCζ mRNA concentration was 0.1 ng/µl for mouse oocyte activation, which was ten times stronger than mPLCζ mRNA. The concentration did not affect parthenogenetic embryo development in vitro. Relatively normal blastocysts were obtained with the same developmental rate (52–53% or 48–51%, respectively) when inactive spermatozoa were injected into activated oocytes using ePLCζ or mPLCζ mRNA injection. However, the birth rate after embryo transfer was slightly but significantly decreased in oocytes activated by ePLCζ mRNA (24%) compared to mPLCζ mRNA (37%) or strontium treatment (40%) activation. These results suggest that the higher activation rate does not always correlate the higher birth rate, and some mechanisms might exist in the oocyte activation process that could affect the later developmental stages like full-term development.https://www.jstage.jst.go.jp/article/jrd/66/1/66_2019-043/_pdf/-char/enhorseinactivated spermatozoamouseoocytes activationsperm-specific phospholipase c zeta (plcζ) |
| spellingShingle | Yunosuke YAMAMOTO Naoki HIROSE Satoshi KAMIMURA Sayaka WAKAYAMA Junya ITO Masatoshi OOGA Teruhiko WAKAYAMA Production of mouse offspring from inactivated spermatozoa using horse PLCζ mRNA The Journal of Reproduction and Development horse inactivated spermatozoa mouse oocytes activation sperm-specific phospholipase c zeta (plcζ) |
| title | Production of mouse offspring from inactivated spermatozoa using horse PLCζ mRNA |
| title_full | Production of mouse offspring from inactivated spermatozoa using horse PLCζ mRNA |
| title_fullStr | Production of mouse offspring from inactivated spermatozoa using horse PLCζ mRNA |
| title_full_unstemmed | Production of mouse offspring from inactivated spermatozoa using horse PLCζ mRNA |
| title_short | Production of mouse offspring from inactivated spermatozoa using horse PLCζ mRNA |
| title_sort | production of mouse offspring from inactivated spermatozoa using horse plcζ mrna |
| topic | horse inactivated spermatozoa mouse oocytes activation sperm-specific phospholipase c zeta (plcζ) |
| url | https://www.jstage.jst.go.jp/article/jrd/66/1/66_2019-043/_pdf/-char/en |
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