Visualization of germ line stem cells in fish

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K22588
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0603:Forestry and forest products science, applied aquatic science, and related fields
• Research Institution: Tokyo University of Marine Science and Technology
• Principal Investigator: ichida kensuke 東京海洋大学, 学術研究院, 助教 (70882637)
• Project Period (FY): 2020-09-11 – 2022-03-31
• Keywords: A型精原細胞 / 生殖細胞移植 / 細胞表面抗原 / 魚類

Outline of Final Research Achievements

To establish the visualization techniques for ASGs aiming for spreading of surrogate broodstock, the identification of the epitope of antibody which are specifically recognize cell surface antigen of ASGs, were conducted. Firstly, we attempted to isolate the antibody-positive protein by 2D electrophoresis with western blotting. As a result, 2 antibody-positive spots could be isolated. Next, mass spectrometry were conducted against these extracted antibody-positive gel-fractions. Consequently, 26 proteins out of 172 total proteins possessing transmembrane regions via in silico analysis. Further, to screen the identification of genes which specifically express in ASGs and not express the other kinds of testicular cells, RT-PCR analysis and in situ hybridization were conducted. As a result, 1 gene were specifically expressed tuna ASGs indicating the possibility that it is the epitope of this antibody.

Free Research Field

発生工学

Academic Significance and Societal Importance of the Research Achievements

本研究で魚類のA型精原細胞の細胞表面に特異的に発現しているタンパク質の同定に成功し、かつ本研究室が保有するA型精原細胞特異抗体の抗原である可能性が強く示唆された。本タンパクが当該抗体の抗原であると確認されれば、これは魚類において抗原として使用可能な分子を初めての同定したことになり、あらゆる魚種において容易にA型精原細胞の細胞操作を可能とするツールとなると期待できる。この4-5年間で生殖細胞移植に関する研究発表数は急増しており、本技術の水産、保全生物学の分野における貢献が世界的に着目されている。そのような背景の中、上述のようにあらゆる魚種に応用可能な細胞操作技術の波及効果は計り知れない。