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Functional analysis of PLC zeta using transgenic sperm.

Research Project

Project/Area Number 18K09144
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 56030:Urology-related
Research InstitutionNagoya City University

Principal Investigator

Kubota Hiroki  名古屋市立大学, 医薬学総合研究院(医学), 研究員 (10347403)

Co-Investigator(Kenkyū-buntansha) 安井 孝周  名古屋市立大学, 医薬学総合研究院(医学), 教授 (40326153)
梅本 幸裕  名古屋市立大学, 医薬学総合研究院(医学), 教授 (80381812)
岩月 正一郎  名古屋市立大学, 医薬学総合研究院(医学), 助教 (70595397)
野崎 哲史  名古屋市立大学, 医薬学総合研究院(医学), 研究員 (50813432)
Project Period (FY) 2018-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2018: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
KeywordsPLC zeta / egg activation / transgenic sperm / エレクトロポレーション / 精巣内遺伝子導入 / Phospholipase C zeta
Outline of Final Research Achievements

We employed a vector designed to produce a fusion of full length clone of mouse PLCζand the enhanced green fluorescent protein. Male mice (4-6 weeks of age) were used for in vivo experiments. For each animal, the right testis was exposed and the PLCζ-GFP injected into the rete testis via the efferent duct. An electrical current was subsequently applied to the right testis. We also detected GFP fluorescence in the epididymal sperm at 2 weeks after the procedure.
These transgenic sperm were introduced into the mouse egg using intracytoplasmic sperm injection technique. We demonstrated the significant elevation of Ca concentration in the egg cytoplasm.

Academic Significance and Societal Importance of the Research Achievements

本研究において、私たちが開発した精巣内遺伝子導入技術は、標的とする遺伝子を導入したtransgenic spermを作成することが可能である。これを応用することにより、標的遺伝子の機能解析並びに受精のメカニズムの解明に多大な貢献を果たすことが期待される。
最終的には遺伝子治療がなされた精子を用いて、遺伝子異常を次世代に継承させない理想的な治療法の開発を目指している。

Report

(5 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • 2019 Research-status Report
  • 2018 Research-status Report

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Published: 2018-04-23   Modified: 2023-01-30  

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