Analysis of Amyloid Related Proteins with Advanced Laser Microdissection

2017 Fiscal Year Final Research Report

• Project/Area Number: 26670152
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Pathological medical chemistry
• Research Institution: Shinshu University
• Principal Investigator: Higuchi Keiichi 信州大学, 学術研究院医学系, 教授 (20173156)
• Co-Investigator(Kenkyū-buntansha): 八谷 如美 地方独立行政法人東京都立産業技術研究センター, 開発本部開発第二部バイオ応用技術グループ, 主任研究員 (30408075) ; 澤下 仁子 信州大学, 学術研究院医学系, 助教 (40359732) ; 矢崎 正英 信州大学, 学術研究院保健学系, 教授 (70372513)
• Research Collaborator: KAMETANI Fuyuki 公益財団法人東京都医学総合研究所, 認知症・高次脳機能研究分野, 主席研究員 (70186013) ; ISHIKAWA Eri 信州大学, 基盤研究支援線ンター機器分析支援部門, 技術職員 ; SUZUKI Kayo 信州大学, 基盤研究支援線ンター機器分析支援部門, 技術職員 ; KAMETANI Kiyokazu 信州大学, 基盤研究支援線ンター機器分析支援部門, 研究支援推進員
• Project Period (FY): 2014-04-01 – 2018-03-31
• Keywords: アミロイドーシス / プロテオーム解析 / バイオ関連機器 / レーザーマイクロダイセクション / 実験病理学 / ApoA-II / 線維蛋白質 / 凝集体

Outline of Final Research Achievements

Proteomic analysis and determination of amyloid inducing activity of fine and small parts of tissue sections were performed using Advanced Laser Microdissection (ALMD) system which were developed recently. Small parts with or without AApoAII amyloid deposition were removed from mouse liver sections with ALMD and contained proteins were extracted. Proteomic analysis with LC-MS/MS system detected major amyloid related proteins. Extracted proteins were injected into recipient mice and induced amyloidosis. In addition, proteomic analysis of amyloid fibril factions of mouse liver indicated the involvement of lipoproteins in the pathogenesis of amyloidosis. Proteomic analysis of small parts of tissue sections of patients containing amyloid deposits collected with ALMD successfuly determined amyloid proteins.
We need further improvement in detection sensitivity and more thorough analysis for elucidating the relationship between protein profiles and seeding activity in amyloidosis.

Free Research Field

病態医化学