Tat-protein disulfide-isomerase A3: a possible candidate for preventing ischemic damage in the spinal cord

Abstract

In the present study, we searched for possible candidates that can prevent ischemic damage in the rabbit spinal cord. For this study, we used two-dimensional gel electrophoresis followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, in sham- and ischemia-operated animals. As the level of protein disulfide-isomerase A3 (PDIA3) significantly decreased 3 h after ischemia/reperfusion, we further investigated its possible role against ischemic damage using an in vitro spinal cord cell line and in vivo spinal cord ischemic model. The administration of Tat-PDIA3 significantly reduced the hydrogen peroxide-induced formation of reactive oxygen species and cell death, based on terminal deoxynucleotidyl transferase-mediated biotinylated dUTP nick end labeling and a colorimetric WST-1 assay. Further, Tat-PDIA3 significantly ameliorated the ischemia-induced deficits in motor function, based on Tarlov’s criteria, 24–72 h after ischemia/reperfusion, as well as the degeneration of motor neurons in the ventral horn 72 h after ischemia/reperfusion. Tat-PDIA3 administration also reduced the ischemia-induced activation of microglia and lipid peroxidation in the motor neurons 72 h after ischemia/reperfusion. PDIA3 also potentially ameliorated the ischemia-induced increase in oxidative markers in serum and decreased the activity of Cu,Zn-superoxide dismutase, Mn-superoxide dismutase, and glutathione peroxidase in spinal cord homogenates, 24 h and 72 h after ischemia/reperfusion. These results suggest that Tat-PDIA3 could be used to protect spinal cord neurons from ischemic damage, due to its modulatory action on the oxidative/anti-oxidative balance. Tat-PDIA3 could be applicable to protects neurons from the ischemic damage induced by thoracoabdominal aorta obstruction.

Differential roles of exogenous protein disulfide isomerase A3 on proliferating cell and neuroblast numbers in the normal and ischemic gerbils

“…In the present study, we focused upon the effects of exogenous PDIA3 on the proliferating cells and neuroblast numbers in the hippocampus, which is one of neurogenic regions in adult brain. First of all, we confirmed the efficient delivery of Tat-PDIA3 in the DG by immunohistochemistry for polyhistidine because we added polyhistidine tag in the Tat-PDIA3 vector as shown in the previous studies (Yoo et al, 2017(Yoo et al, , 2019. Tat-PDIA3-treated control and ischemic groups showed strong polyhistidine immunoreactivity in the DG, while vehicle-treated control and ischemic groups did not show any prominent polyhistidine immunoreactivity in the hippocampal DG.…”

“…In previous studies, we generated Tat‐PDIA3 fusion protein and observed the intracellular delivery of Tat‐PDIA3 fusion protein (Yoo et al, , ) as well as the colocalization of PDIA3 with ER tracker (Yoo et al, ). In addition, we found the neuroprotective actions of Tat‐PDIA3 and its mechanisms in the gerbil brain ischemia (Yoo et al, ) and rabbit spinal cord ischemia (Yoo et al, ). Especially, administration of PDIA3 protected neurons from hydrogen peroxide‐induced oxidative stress in HT22 cells and ischemic damage in the gerbil hippocampal CA1 region (Yoo et al, ).…”

“…Human immunodeficiency virus-1 (HIV-1) Tat would be one of best strategy for cellular delivery into central nervous system because they efficiently transport peptide or proteins which have higher molecular weights than their own (Van den Berg & Dowdy, 2011) because the efficient transduction capability has been confirmed using various peptides or proteins against ischemic damage (Jo et al, 2017;Tu et al, 2015;Zhou et al, 2015). In previous studies, we generated Tat-PDIA3 fusion protein and observed the intracellular delivery of Tat-PDIA3 fusion protein (Yoo et al, 2017(Yoo et al, , 2019 as well as the colocalization of PDIA3 with ER tracker (Yoo et al, 2017). In addition, we found the neuroprotective actions of Tat-PDIA3 and its mechanisms in the gerbil brain ischemia (Yoo et al, 2019) and rabbit spinal cord ischemia (Yoo et and Cell Biology, College of Veterinary Medicine, Seoul National University, Seoul 08826, South Korea.…”

Effects of Pyridoxine Deficiency on Hippocampal Function and Its Possible Association with V-Type Proton ATPase Subunit B2 and Heat Shock Cognate Protein 70

“…Animals (n = 20 in each group) were anesthetized with a mixture of alfaxalone and xylazine at 8 weeks after diet feeding and the hippocampus was isolated from the whole brain. Hippocampal tissues were suspended in a sample buffer [29], which consisted of 30 mM Tris, 7 M urea, 2 M thiourea, 65 mM dithiothreitol (DTT), 4% 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) with 40 µL protease inhibitor (pH 8.5). Suspensions were sonicated five times for 10 sec and centrifuged at 45,000 rpm for 45 min.…”

2D Gel Electrophoresis to Address Biological Issues

“…The main advantage of 2DE is the capability of simultaneously resolving complex protein mixture, up to 10,000, allowing their quantitation as well as the analysis of all post-translational modifications [5]. 2DE has been adopted for a variety of applications: from drug discovery to diagnostics, therapy, and biochemistry [6][7][8][9][10].…”