Oxidative Stress Induction of DJ‐1 Protein in Reactive Astrocytes Scavenges Free Radicals and Reduces Cell Injury

Takashi Yanagida,Hiroyoshi Ariga,Tomonori Shibaike,Toshihiro Inubushi,Ikuo Tooyama,Takahiro Taira,Shigehiro Morikawa,Atsuko Yamamoto,Yoshihisa Kitamura,Kazuyuki Takata,Jun Tsushima,Takashi Taniguchi,Hiroyuki Yasui,Daijiro Yanagisawa

doi:10.4161/oxim.2.1.7985

Abstract

Astrocytes, one of the predominant types of glial cells, function as both supportive and metabolic cells for the brain. Under cerebral ischemia/reperfusion-induced oxidative conditions, astrocytes accumulate and activate in the ischemic region. DJ-1 has recently been shown to be a sensor of oxidative stress in living cells. However, the function of astrocytic DJ-1 is still unknown. In the present study, to clarify the effect of astrocytic DJ-1 protein under massive oxidative insult, we used a focal ischemic rat model that had been subjected to middle cerebral artery occlusion (MCAO) and reperfusion. We then investigated changes in the distribution of DJ-1 in astrocytes, DJ-1 release from cultured astrocytes, and the effects of recombinant DJ-1 protein on hydrogen peroxide (H(2)O(2))-induced death in normal and DJ-1-knockdown SH-SY5Y cells and on in vitro scavenging of hydroxyl radicals ((*)OH) by electron spin resonance spectrometry. At 24 h after 2-h MCAO and reperfusion, an infarct lesion was markedly observed using magnetic resonance imaging and 2,3,5-triphenyltetrazolium chloride staining. In addition, reactive astrocytes enhanced DJ-1 expression in the penumbral zone of the ischemic core and that DJ-1 protein was extracellularly released from astrocytes by H2O2 in in vitro primary cultures. Although DJ-1-knockdown SH-SY5Y cells were markedly vulnerable to oxidative stress, treatment with glutathione S-transferase-tagged recombinant human DJ-1 protein (GST-DJ-1) significantly inhibited H(2)O(2)-induced cell death. In addition, GST-DJ-1 protein directly scavenged (*)OH. These results suggest that oxidative stress induces the release of astrocytic DJ-1 protein, which may contribute to astrocyte-mediated neuroprotection.

Highlights

DJ-1, which was discovered as a novel oncogene product in collaboration with activated small GTP-binding protein ras,[1] has recently been identified as the causative gene of familial Parkinson’s disease PARK7.2 DJ-1 has many functions, such as transcriptional factor, molecular chaperone, etc.[3,4,5,6] X-ray crystallographic and biological analyses have shown that wild-type DJ-1 protein forms a homodimer
glutathione S-transferase (GST)-DJ-1 protein directly scavenged OH. These results suggest that oxidative stress induces the release of astrocytic DJ-1 protein, which may contribute to astrocyte-mediated neuroprotection
To analyze changes in DJ-1 in the brain under oxidative stress, we used an ischemic rat model that had been subjected to middle cerebral www.landesbioscience.com artery occlusion (MCAO) and reperfusion

Summary

Introduction

DJ-1, which was discovered as a novel oncogene product in collaboration with activated small GTP-binding protein ras,[1] has recently been identified as the causative gene of familial Parkinson’s disease PARK7.2 DJ-1 has many functions, such as transcriptional factor, molecular chaperone, etc.[3,4,5,6] X-ray crystallographic and biological analyses have shown that wild-type DJ-1 protein forms a homodimer. One of the leading causes of death and longlasting disability, results from a transient or permanent reduction in Oxidative Medicine and Cellular Longevity

Methods

Results

Conclusion