FullText URL Autophagy_6_8_1107-1114.pdf
Author Tian, FengFeng| Deguchi, Kentaro| Yamashita, Toru| Ohta, Yasuyuki| Morimoto, Nobutoshi| Shang, Jingwei| Zhang, Xuemei| Liu, Ning| Ikeda, Yoshio| Matsuura, Tohru| Abe, Koji|
Abstract 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.
Keywords autophagy apoptosis GFP-LC3 Tg mice in vivo imaging tMCAO
Published Date 2010-11-16
Publication Title Autophagy
Volume volume6
Issue issue8
Start Page 1107
End Page 1114
ISSN 1554-8627
Content Type Journal Article
language 英語
OAI-PMH Set 岡山大学
File Version author
PubMed ID 20930570
DOI 10.4161/auto.6.8.13427
Web of Sience KeyUT 000284258900010
Official Url http://doi.org/10.4161/auto.6.8.13427|
Related Url http://ousar.lib.okayama-u.ac.jp/metadata/49128