フルテキストURL Autophagy_6_8_1107-1114.pdf
著者 Tian, FengFeng| Deguchi, Kentaro| Yamashita, Toru| Ohta, Yasuyuki| Morimoto, Nobutoshi| Shang, Jingwei| Zhang, Xuemei| Liu, Ning| Ikeda, Yoshio| Matsuura, Tohru| Abe, Koji|
抄録 Recent studies have suggested that autophagy is involved in a neural death pathway following cerebral ischemia. In vivo detection of autophagy could be important for evaluating ischemic neural cell damage for human stroke patients. Using novel green fluorescent protein (GFP)-fused microtubule-associated protein 1 light chain 3 (LC3) transgenic (Tg) mice, in vivo imaging of autophagy was performed at 1, 3 and 6 d after 60 min transient middle cerebral artery occlusion (tMCAO). Ex vivo imaging of autophagy, testing of the autophagy inhibitor 3-methyladenine (3-MA), estern blot analysis, immunohistochemistry, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) and fluorescent analyses were performed on brain sections following tMCAO. In vivo fluorescent signals were detected above the ischemic hemisphere through the skull bone at 1, 3 and 6 d after tMCAO, with a peak at 1 d. Similar results were obtained with ex vivo fluorescence imaging. western blot analysis revealed maximum LC3-I and LC3-II expression at 1 d after tMCAO and fluorescence immunohistochemistry demonstrated that GFP-LC3-positive cells were primarily neuronal, not astroglial or microglial, cells. The number of GFP-LC3/TUNEL double-positive cells was greater in the peri-ischemic area than in the core. These results provided evidence of in vivo autophagy detection, with a peak at 1 d, in a live animal model following cerebral ischemia. This novel technique could be valuable for monitoring autophagic processes in vivo in live stroke patients, as well as for clarifying the detailed role of autophagy in the ischemic brain, as well as in other neurological diseases.
キーワード autophagy apoptosis GFP-LC3 Tg mice in vivo imaging tMCAO
発行日 2010-11-16
出版物タイトル Autophagy
6巻
8号
開始ページ 1107
終了ページ 1114
ISSN 1554-8627
資料タイプ 学術雑誌論文
言語 English
OAI-PMH Set 岡山大学
論文のバージョン author
PubMed ID 20930570
DOI 10.4161/auto.6.8.13427
Web of Sience KeyUT 000284258900010
オフィシャル URL http://doi.org/10.4161/auto.6.8.13427|
関連URL http://ousar.lib.okayama-u.ac.jp/metadata/49128