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Comparison of abnormal isoform of prion protein in prion-infected cell lines and primary-cultured neurons by PrPSc-specific immunostaining

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J.Gen.Virol.v.97p.2030-2042(2016) .pdf10.07 MBPDF見る/開く
この文献へのリンクには次のURLを使用してください:http://hdl.handle.net/2115/66951

タイトル: Comparison of abnormal isoform of prion protein in prion-infected cell lines and primary-cultured neurons by PrPSc-specific immunostaining
著者: Tanaka, Misaki 著作を一覧する
Fujiwara, Ai 著作を一覧する
Suzuki, Akio 著作を一覧する
Yamasaki, Takeshi 著作を一覧する
Hasebe, Rie 著作を一覧する
Masujin, Kentaro 著作を一覧する
Horiuchi, Motohiro 著作を一覧する
キーワード: prions
Immunofluorescence assay
primary neurons
guanidine isothiocyanate
monoclonal antibody
発行日: 2016年 8月
出版者: Society for General Microbiology
誌名: Journal of General Virology
巻: 97
開始ページ: 2030
終了ページ: 2042
出版社 DOI: 10.1099/jgv.0.000514
抄録: We established abnormal isoform of prion protein (PrPSc)-specific double immunostaining using mAb 132, which recognizes aa 119-127 of the PrP molecule, and novel PrPSc-specific mAb 8D5, which recognizes the N-terminal region of the PrP molecule. Using the PrPSc-specific double immunostaining, we analysed PrPSc in immortalized neuronal cell lines and primary cerebral-neuronal cultures infected with prions. The PrPSc-specific double immunostaining showed the existence of PrPSc positive for both mAbs 132 and 8D5, as well as those positive only for either mAb 132 or mAb 8D5. This indicated that double immunostaining detects a greater number of PrPSc species than single immunostaining. Double immunostaining revealed cell-type-dependent differences in PrPSc staining patterns. In the 22 L prion strain-infected Neuro2a (N2a)-3 cells, a subclone of N2a neuroblastoma cell line, or GT1-7, a subclone of the GT1 hypothalamic neuronal cell line, granular PrPSc stains were observed at the perinuclear regions and cytoplasm, whereas unique string-like PrPSc stains were predominantly observed on the surface of the 22 L strain-infected primary cerebral neurons. Only 14% of PrPSc in the 22 L strain-infected N2a-3 cells were positive for mAb 8D5, indicating that most of the PrPSc in N2a-3 lack the N-terminal portion. In contrast, nearly half PrPSc detected in the 22 L strain-infected primary cerebral neurons were positive for mAb 8D5, suggesting the abundance of full-length PrPSc that possesses the N-terminal portion of PrP. Further analysis of prion-infected primary neurons using PrPSc-specific immunostaining will reveal the neuron-specific mechanism for prion propagation.
資料タイプ: article (author version)
URI: http://hdl.handle.net/2115/66951
出現コレクション:雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

提供者: 堀内 基広

 

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