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Please use this identifier to cite or link to this item: http://hdl.handle.net/10119/14330

Title: プラズモンイメージングとサブ細胞レベル磁気分離によるオートファジーの理解
Other Titles: Understanding of autophagy utilizing plasmon imaging and subcellular magnetic separation techniques
Authors: 前之園, 信也
Authors(alternative): Maenosono, Shinya
Keywords: オートファジー
Issue Date: 1-Jun-2017
Abstract: 磁性-プラズモンハイブリッドナノ粒子(Ag/FeCo/Agコア/シェル/シェル型ナノ粒子)を創製し、このナノ粒子を用いてオートファゴソームの磁気分離に挑戦した。ハイブリッドナノ粒子を哺乳細胞にリポフェクションし培養したところ、30分後にはナノ粒子がVps26と共局在し、その後LC3と共局在する様子が観察された。最適なタイミングで細胞膜を温和に破砕し磁気分離に供した。磁気分離分画にはLC3-II、トランスフェリン受容体、及びLAMP2が濃縮されていたが、LC3-Iは含有されていなかった。これらの結果はオートファゴソームが単離できたことを示している。:We developed ultrasmall magnetic-plasmonic hybrid nanobeads and applied them to the isolation of autophagosomes by applying a magnetic field. The beads were chemically synthesized and comprised an Ag/FeCo/Ag core/shell/shell structure with a mean diameter of 15 nm. The Ag core and the FeCo shell conferred imaging and magnetic separation capabilities, respectively. The nanobeads were transfected into mammalian cells by lipofection. Thirty minutes after lipofection, the nanobeads co-localized with Vps26, and subsequently with LC3. Cell lysates were prepared at the appropriate time points and were subjected to magnetic separation. The separated fraction contained LC3-II, transferrin receptor, and LAMP2, but not LC3-I, suggesting that autophagosomes of endosomal origin had been isolated.
Description: 挑戦的萌芽研究
Language: jpn
URI: http://hdl.handle.net/10119/14330
Appears in Collections:平成28年度 (FY 2016)

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