Genetic and single-cell approaches to evaluate roles played by UPIII-Src system in frog oocyte maturation and egg fertilization

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K07083
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Developmental biology
• Research Institution: Kyoto Sangyo University
• Principal Investigator: SATO Ken-ichi 京都産業大学, 総合生命科学部, 教授 (30235337)
• Research Collaborator: TOKMAKOV Alexander A 京都産業大学, 総合生命科学部, 研究助教 (20301278) ; NAKAJIMA Keisuke 広島大学, 両生類研究センター, 助手 (60260311) ; IJIRI Takashi 摂南大学, 理工学部生命科学科, 講師 (20629620)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 受精 / 卵成熟 / シグナル伝達 / チロシンリン酸化 / 細胞膜マイクロドメイン / ウロプラキンIII / ゲノム編集 / Src

Outline of Final Research Achievements

We tried to prepare a uroplakin III (UPIII)-null oocyte and egg with the use of Xenopus tropicalis as a model animal, and of genome editing technologies such as TALENs and CRISPR/Cas9. At present, however, because of the highly unstable viability in UPIII-homozygously negative larva, we have not yet succeeded in obtaining females with homologous disruption in the UPIII gene loci. Therefore, we are still in the process of preparing a homozygous UPIII-null female. Under this condition, a single-cell biology approach toward oocytes and eggs that can be obtained from heterozugous UPIII-null females, such as indirect immunofluorescent experiments with the use of anti-UPIII antibody, and in vitro reconstitution of sperm-induced membrane-associated signaling events (e.g. tyrosine phosphorylation of Src) are under investigation.

Free Research Field

発生生物学