Expression and localization of Fas-associated proteins following focal cerebral ischemia in rats.
The aim of this study was to investigate the changes of expression of Fas-associated proteins and its cellular localization in the peri-infarct region following transient focal cerebral ischemia. Adult male Sprague-Dawley rats underwent right middle cerebral artery occlusion (MCAo) for 2 h and reperfusion for 1, 3, 6, 12 and 24 h. The expression of Fas-associated death domain protein (FADD), Fas-associated phosphatase-1 (FAP-1) caspase-8 and death-associated protein (Daxx), the pro-apoptotic genes, were examined by methods of RT-PCR, immunohistochemistry and Western blot. The results showed that the expression levels of mRNA and protein for FADD and caspase-8 increased significantly at 1-3 h after reperfusion, peaked at 12 h, then declined markedly at 24 h. The time course change of FAP-1 was consistent with that of FADD. The expression level of mRNA and protein for death-associated protein (Daxx) increased significantly at 3 h after reperfusion and persisted for 24 h at a high level. Immunofluorescence double-staining laser scanning showed that the immunoreactivity of FADD was localized in cytoplasm, and Daxx immunoreactivity was translocated from nucleus to cytoplasm at 3 h after reperfusion. The TUNEL-positive cells could be found in peri-infarct region at 3 h and increased with time after reperfusion. Our findings suggest a possible association between expression of FADD, caspase-8, Daxx and FAP-1 genes and apoptosis following ischemia.
Bi
Fang-fang
FF
Department of Neurology, Xiangya Hospital, Central South University, Changsha, Hunan 410008, China. bi000010@flinders.edu.au
Xiao
Bo
B
Hu
Yue-qiang
YQ
Tian
Fa-fa
FF
Wu
Zhi-guo
ZG
Ding
Ling
L
Zhou
Xin-fu
XF
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis / Q000235:genetics / Q000502:physiology; D001930:Brain Injuries / Q000209:etiology / Q000378:metabolism; D002545:Brain Ischemia / Q000150:complications / Q000378:metabolism*; D053181:Caspase 8 / Q000235:genetics / Q000378:metabolism; D053200:Fas-Associated Death Domain Protein / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D008297:Male; D018832:Molecular Chaperones; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D054595:Protein Tyrosine Phosphatase, Non-Receptor Type 13 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000502:physiology*; D013997:Time Factors; D014018:Tissue Distribution
D048868:Adaptor Proteins, Signal Transducing; C528558:Daxx protein, rat; D053200:Fas-Associated Death Domain Protein; D018832:Molecular Chaperones; D009687:Nuclear Proteins; D012333:RNA, Messenger; D054595:Protein Tyrosine Phosphatase, Non-Receptor Type 13; C528870:Ptpn13 protein, rat; D053181:Caspase 8