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Noninvasive Measurement and Tracing Methodology

開催日 2014/9/11
時間 14:00 - 15:00
会場 Room H(304)
Chairperson(s) 澁木 克栄 / Katsuei Shibuki (新潟大学脳研究所システム脳生理学分野 / Brain Research Institute, Niigata University, Japan)
西村 幸男 / Yukio Nishimura (生理学研究所・発達生理学研究系 認知行動発達機構研究部門 / Department of Developmental Physiology, Division of Behavioral Development, NATIONAL INSTITUTE for PHYSIOLOGICAL SCIENCES, Japan)

Three orthogonal Gate-Tag vectors for pathway-specific gene transfer in the mammalian brain

  • O1-H-3-4
  • 松山 真 / Makoto Matsuyama:1,2 大橋 陽平 / Yohei Ohashi:1 坪田 匡史 / Tadashi Tsubota:1 矢口 雅江 / Masae Yaguchi:1 眞々田 薫里 / Kaori Mamada:1 加藤 成樹 / Shigeki Kato:3 小林 和人 / Kazuto Kobayashi:3 宮下 保司 / Yasushi Miyashita:1 
  • 1:東京大院医統合生理 / Dept. of Physiol., Univ. of Tokyo Sch. of Med., Tokyo, Japan 2:日本学術振興会特別研究員 / JSPS Research Fellow, Tokyo, Japan 3:福島県立医大医生体情報伝達研生体機能 / Dept. of Molecular Genetics, Inst. of Biomedical Sciences, Fukushima Med. Univ., Fukushima, Japan 

The development of new imaging and stimulating techniques is facilitating the analysis of complex neural systems, but the conventional genetic tools do not yet allow the separate and simultaneous manipulation of multiple neural pathways in brain regions containing neurons projecting to different target regions. To untangle complex neural circuits, we have developed a new viral gene transfer technique termed the "Gate-Tag vector system". This vector system is composed of two elements: a Gate vector that expresses a specific receptor "Gate" on the target cell surface, and a Tag vector that can entry target cells via the specific Gate. This system effectively creates a door on a cell's surface through which a selective Tag vector can be introduced into the cell. To implement this system, we utilized three avian-derived receptors as gates (Gates A-C) and three types of avian sarcoma and leukosis virus (ASLV) envelopes as tags (Tags A-C). When the lentiviral vectors pseudotyped with ASLV envelopes were used to infect receptor-expressing cell populations, substantially high receptor-envelope selectivity was observed both in vitro (>97% in HEK293T cells) and in vivo (>98% in the rat brains). By targeting the expression of each type of receptors with highly efficient retrograde gene transfer (HiRet), we achieved pathway-specific, differential fluorescent labeling of three thalamic neuronal populations, each projecting to different cortical regions. This proof-of-concept study demonstrates that our novel multi-target gene transfer system can genetically dissociate intermingled neural pathways in vivo.

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