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KAKEN_16K13107seika.pdf
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Download
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:Nov 12, 2018 |
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Total downloads since Nov 12, 2018 : 616
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シナプス分子のスプライスバリアント特異的な局在を検出するための技術開発
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シナプス ブンシ ノ スプライス バリアント トクイテキナ キョクザイ オ ケンシュツスル タメ ノ ギジュツ カイハツ
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Shinapusu bunshi no supuraisu barianto tokuitekina kyokuzai o kenshutsusuru tame no gijutsu kaihatsu
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Application of epitope-tag insertion for the analysis of synaptic molecules splicing isoform expressions in the brain
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井端, 啓二
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イバタ, ケイジ
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Ibata, Keiji
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慶應義塾大学・医学部(信濃町)・助教
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Research team head
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科研費研究者番号 : 30462659
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柚崎, 通介
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ユザキ, ミチスケ
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Yuzaki, Michisuke
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慶應義塾大学・医学部(信濃町)・教授
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Research team member
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科研費研究者番号 : 40365226
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竹尾, ゆかり
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タケオ, ユカリ
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Takeo, Yukari
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慶應義塾大学・医学部(信濃町)・特任助教
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Research team member
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科研費研究者番号 : 90624320
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林, 亜由美
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ハヤシ, アユミ
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Hayashi, Ayumi
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2018
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科学研究費補助金研究成果報告書
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2017
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Cbln1は小脳におけるシナプス形成分子であり, 他の脳領域ではCbln2および4も重要な働きをしていると考えられているが局在は不明である。Cbln分子はスプライスサイト4を持つNeurexin(NrxSS4)に結合するが, NrxSS4の個々のシナプスでの局在に関しても, 特異的な抗体が存在しないため不明であった。本研究では, 抗体を作る代わりに, ゲノム編集技術を用いてエピトープタグそのものをノックインすることで, Cbln分子とNrxSS4の局在を解明する事を試みた。その結果Cbln1およびCbln2のノックインマウスは得られたが, NrxSS4では得られなかった。現在免疫組織実験を進行中である。
Cbln1 induces synapse formation in the central nervous system, especially in the cerebellum. In other brain region, it is speculated that other Cbln proteins, Cbln2 and 4, also plays an important role at synapse formations. It is known Cbln proteins bind with splice site 4 containing neurexin isoform at presynaptic site. To understand the characteristic of each synapse in the brain, it is important to know which Cbln proteins and which neurexin isoform is expressed at each presynaptic site. To date, there are no specific antibody for neurexin splice site 4 and for Cbln family proteins because of high-similarities. To overcome this difficulty, we took advantage of epitope-tag and antibody detection system. We tried to insert epitope-tag at splice site 4 of neurexin and at terminus of Cbln proteins using CRISPR/Cas9 system. In this research period, we found difficulty to get knock-in mouse for neurexin. On the other hand, we obtained epitope-tag inserted Cbln1 and Cbln2 knock-in mice.
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研究種目 : 挑戦的萌芽研究
研究期間 : 2016～2017
課題番号 : 16K13107
研究分野 : 神経科学
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