2020 Fiscal Year Annual Research Report
JUNO mRNA補完法による精子卵融合の分子機構解明
| Project/Area Number |
20J10363
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| Research Institution | Osaka University |
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Principal Investigator |
PARK SOOJIN 大阪大学, 医学系研究科, 特別研究員(DC2)
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| Project Period (FY) |
2020-04-24 – 2022-03-31
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| Keywords | sperm mitochondria |
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| Outline of Annual Research Achievements |
I wanted to reveal the mechanism of how and when JUNO (fusion protein in egg membrane) is cleaved. For that, I synthesized a transmembrane type of Juno mRNA, which is originally GPI-anchored protein in vivo. With this, I tried to prove that JUNO does not disappear because it is a transmembrane protein, and as a result, a maximum number of sperm will fuse. I tested this by mRNA injecting into mouse GV stage oocyte. As a result, only 0.13 sperm in average fused with 11.2% of mRNA injected egg, while 1.05 sperm fused with 67.9% of control egg. We concluded that this mRNA does not work well, and better to generate a trans-gene(TG)mice using a special promoter for oocyte-specific expression. The TG mice generation may take long time, so I decided to analyze testis-specific gene knockout(KO)mice.
I analyzed 10 KO male mice which could produce pups normally. With these KO mice, I could report that those 10 genes are not essential and thus cannot be used as a candidate gene for further infertility studies. （Biol Reprod. 2020 Aug 4;103(2):195-204）
Besides, I found Tbc1d21 KO male mice could not produce pups. KO sperm had abnormal mitochondrial sheath in midpiece, and thus sperm motility was too low to reach ovulated eggs at last. Meanwhile, Armc12 KO had similar phenotype. By immunoprecipitation, we concluded that TBC1D21 and ARMC12 works together in testis and participate in the formation of mitochondrial sheath of sperm. I could show the data of Tbc1d21 KO mice in the following paper. （Proc Natl Acad Sci U S A. 2021 Feb 9;118(6):e2018355118）
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| Research Progress Status |
翌年度、交付申請を辞退するため、記入しない。
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| Strategy for Future Research Activity |
翌年度、交付申請を辞退するため、記入しない。
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Research Products
(3 results)
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[Journal Article] CRISPR/Cas9-mediated genome-edited mice reveal 10 testis-enriched genes are dispensable for male fecundity2020
Author(s)
Soojin Park, Keisuke Shimada, Yoshitaka Fujihara, Zoulan Xu, Kentaro Shimada, Tamara Larasati, Putri Pratiwi, Ryan M Matzuk, Darius J Devlin, Zhifeng Yu, Thomas X Garcia, Martin M Matzuk, Masahito Ikawa.
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Journal Title
Biology of Reproduction
Volume: 103
Pages: 195-204
DOI
Peer Reviewed / Open Access / Int'l Joint Research
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