Neuroprotective effects of PEP-1-carbonyl reductase 1 against oxidative-stress-induced ischemic neuronal cell damage

Tat-PRAS40 prevent hippocampal HT-22 cell death and oxidative stress induced animal brain ischemic insults

“…For histological analysis, gerbils were sacrificed 7 days after ischemiareperfusion and brain tissues were extracted. Then the brain tissues were cryoprotected, frozen, sectioned (50 μm), and immunostaining was performed as previously described [21,24,37]. …”

Tat-NOL3 protects against hippocampal neuronal cell death induced by oxidative stress through the regulation of apoptotic pathways

“…In previous studies, we demonstrated that transduced PTD fusion proteins protect against delayed pyramidal neuronal degeneration in the hippocampal CA1 region in an animal model of ischemia (23,33). Thus, the protective effects of Tat-NOL3 protein against ischemic damage were determined by immunohistochemistry.…”

“…To examine Δ Ψm , 14 µM Tat-NOL3 protein or 14 µM con-NOL3 protein was transduced into the HT22 cells for 1 h and subsequently, they were exposed to 0.5 mM H 2 O 2 for 2 h. Δ Ψm was assessed as described previously (33,34). Briefly, 100 µl of JC-1 staining solution per ml of culture medium was added and the cells were incubated in a CO 2 incubator at 37˚C for 15 min.…”

“…To examine the effects of Tat-NOL3 on ischemic damage, brain tissue samples were obtained at 7 days after ischemia-reperfusion. Immunohistochemistry was performed as described previously (33,35,36). The animals were anesthetized with 30 mg/kg Zoletil 50 (Virbac Laboratories, Carros, France) at 7 days after ischemia-reperfusion and perfused transcardially with 0.1 M PBS (pH 7.4) followed by 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4).…”

“…They were then exposed to biotinylated rabbit anti-goat IgG (1:200; Vector Laboratories, Inc., Burlingame, CA, USA) or goat antimouse IgG and streptavidin-peroxidase complex and visualized with 3,3'-diaminobenzidine (Sigma-Aldrich, St. Louis, MO, USA). Cresyl violet (CV; Junsei Chemical Co. Ltd., Saitama, Japan) and Fluoro-Jade B (FJB; Millipore Co., Temecula, CA, USA) staining were performed as previously described (23,33). Additionally, we performed immunostaining with a rabbit anti-histidine antibody and DAPI to detect the transduction of Tat-NOL3 protein in the brain hippocampus.…”