Expression and localization of Fas-associated proteins following focal cerebral ischemia in rats.

Abstract

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.

• Authors: Fang-fang Bi, Bo Xiao, Yue-qiang Hu, Fa-fa Tian, Zhi-guo Wu, Ling Ding, Xin-fu Zhou
• Journal: Brain research (Brain Res) Vol. 1191 Pg. 30-8 (Jan 29 2008) ISSN: 0006-8993 [Print] Netherlands
• PMID: 18096138 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Adaptor Proteins, Signal Transducing
• Daxx protein, rat
• Fas-Associated Death Domain Protein
• Molecular Chaperones
• Nuclear Proteins
• RNA, Messenger
• Protein Tyrosine Phosphatase, Non-Receptor Type 13
• Ptpn13 protein, rat
• Caspase 8

Topics

• Adaptor Proteins, Signal Transducing (genetics, metabolism)
• Animals
• Apoptosis (genetics, physiology)
• Brain Injuries (etiology, metabolism)
• Brain Ischemia (complications, metabolism)
• Caspase 8 (genetics, metabolism)
• Fas-Associated Death Domain Protein (genetics, metabolism)
• Immunohistochemistry
• Male
• Molecular Chaperones
• Nuclear Proteins (genetics, metabolism)
• Protein Tyrosine Phosphatase, Non-Receptor Type 13 (genetics, metabolism)
• RNA, Messenger (analysis)
• Random Allocation
• Rats
• Rats, Sprague-Dawley
• Signal Transduction (physiology)
• Time Factors
• Tissue Distribution