Rescue of Neuronal Cell Death by ER-stress protein over expression

ER应激蛋白过度表达拯救神经细胞死亡

• 批准号: 14580725
• 负责人: KITAO Yasuko
• 金额: $ 2.3万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580725/
• 关键词: Astrocytes; Brain ischemia; ER-stress; Gene Therapy; Glutamic acid; Ischemic tolerance; 興奮性アミノ酸; 神経細胞死

ORP150 is a novel stress protein localized in the endoplasmic reticulum (ER). To investigate the role of ORP150 in delayed neuronal cell death, we have examined its expression in the gerbil brain after the ischemic insult. The expression of ORP150 antigen, as well as its transcripts, was observed in the CA1 region after the occlusion of the common carotid altety, and this was enhanced by the preconditioning. In cultured neurons, exposure to either hypoxia or glutamate induced the expression of ORP150, and this was also observed by treating the culture with either thapsigargin or breferdin-A, indicating that both glutamate and hypoxia can cause shess in the ER (ER stress). Neurons became more vulnerable to these stresses following treatment of either cyclcheximide or the infection with an adenovirus carrying ORP150 antisense structure. In cantrast, the overexpression of ORP150 by adenovinus suppressed the neuronal cell death, and this was accompanied by the suppression of the Ca2+ eleva … More tion and proteolytic activity induced by glutamate. Further, overexpressicn of ORP150 in CA1 neurons by the adenovirus carrying ORP150-sense struciture suppressed delayed neuronal cell death after ischemia. These data suggest a possible function of ORP150 as an intracellular apparatus, which participates in a protective response in ischemic tolerance.A series of events initiated by glutamate-receptor interaction perturbs cellular homeostasis resulting in elevation of intracellular free calcium and cell death. Cells subject to such environmental change express stress proteins, which contribute importantly to maintenance of metabolic homeostasis and viability. We show that an inducible chaperone present in endoplasmic reticulum (ER), the 150 kDa oxygen-regulated protein (ORP150) is expressed both in human brain after seizure attack and in mice hippocampus after kainite administration. Using mice heterozygous for ORP150 deficiency, exposure to excitatory stimuli caused hippocampal neurons to display exaggerated elevation of cytosolic calcium accompanied by activation of μ-calpain and cathepsin B, as well as increased vulnerability to glutamate-induced cell death in vitro and decreased survival to kainate in vivo. In contrast, targeted neuronal overexpression of ORP150 suppressed each of these events, and enhanced neuronal and animal survival in parallel with diminished seizure intensity. Studies using cultured hippocampal neurons showed that ORP150 regulates cytosolic free calcium and activation of proteolytic pathways causing cell death in neurons subject to excitatory stress. Our data underscore a pivotal role for ER stress in glutamate toxicity, and pinpoint a key ER chaperone, ORP150, which orchestrates the protective stress response critical for neuronal survival. Less

ORP 150是一种新的内质网应激蛋白。为了研究ORP 150在迟发性神经元细胞死亡中的作用，我们检测了其在缺血损伤后沙土鼠脑中的表达。在阻断颈总动脉后，在CA 1区观察到ORP 150抗原及其转录物的表达，并且这种表达被预处理增强。在培养的神经元中，暴露于缺氧或谷氨酸诱导ORP 150的表达，并且这也通过用毒胡萝卜素或breferdin-A处理培养物来观察，表明谷氨酸和缺氧都可以引起ER中的shess（ER应激）。神经元变得更容易受到这些压力治疗后，无论是cyclcheximide或感染携带ORP 150反义结构的腺病毒。在cantrast中，腺病毒介导的ORP 150过表达抑制了神经细胞的死亡，并伴随着Ca 2+升高的抑制。 ...更多信息 谷氨酸诱导的蛋白水解活性。此外，通过携带ORP 150正义结构的腺病毒在CA 1神经元中过表达ORP 150抑制缺血后迟发性神经元细胞死亡。提示ORP 150可能作为一种细胞内装置参与缺血耐受的保护性反应，通过谷氨酸-受体相互作用引发的一系列事件扰乱细胞内稳态，导致细胞内游离钙升高和细胞死亡。受到这种环境变化的细胞表达应激蛋白，其对维持代谢稳态和活力有重要贡献。我们发现，诱导分子伴侣存在于内质网（ER），150 kDa的氧调节蛋白（ORP 150）在癫痫发作后在人脑和红藻氨酸后在小鼠海马中表达。使用ORP 150缺陷杂合子小鼠，暴露于兴奋性刺激导致海马神经元显示胞质钙过度升高，伴随μ-钙蛋白酶和组织蛋白酶B的激活，以及体外谷氨酸诱导细胞死亡的脆弱性增加和体内红藻氨酸诱导细胞存活率降低。相反，靶向神经元过表达ORP 150抑制了这些事件中的每一个，并提高了神经元和动物的存活率，同时降低了癫痫发作强度。使用培养的海马神经元的研究表明，ORP 150调节胞质游离钙和蛋白水解途径的激活，导致神经元受到兴奋性应激的细胞死亡。我们的数据强调了内质网应激在谷氨酸毒性中的关键作用，并确定了一个关键的内质网伴侣蛋白ORP 150，它协调了对神经元存活至关重要的保护性应激反应。少

项目成果

Transmission of cell stress from endoplasmic reticulum to mitochondria: enhanced expression of Lon protease.

• DOI: 10.1083/jcb.200108103
• 发表时间: 2002-06-24
• 期刊: The Journal of cell biology
• 作者: Hori O;Ichinoda F;Tamatani T;Yamaguchi A;Sato N;Ozawa K;Kitao Y;Miyazaki M;Harding HP;Ron D;Tohyama M;M Stern D;Ogawa S
• 通讯作者: Ogawa S

The ER chaperone 150 kDa Oxygen Regulated Protein (ORP150) improves insulin resistance in Type 2 Diabetes Mellitus.

ER 伴侣 150 kDa 氧调节蛋白 (ORP150) 可改善 2 型糖尿病的胰岛素抵抗。

• 发表时间: 2005
• 期刊: Diabetes. 68
• 作者: 小澤;小川;他
• 通讯作者: 他

The ER chaperone 150 kDa Oxygen Regulated Protein (ORP150) improves insulin resistance in Type 2 Diabetes Mellitus. Diabetes.

ER 伴侣 150 kDa 氧调节蛋白 (ORP150) 可改善 2 型糖尿病的胰岛素抵抗。

• 发表时间: 2005
• 期刊: Diabetes. 54
• 作者: 小澤;北尾ほか
• 通讯作者: 北尾ほか

Accumulation of microglial cells expressing ELR motif-positive CXC chemokines and their receptors CXCR2 in monkey hippocanpus after ischemia-reperfusion.

缺血再灌注后猴海马中表达 ELR 基序阳性 CXC 趋化因子及其受体 CXCR2 的小胶质细胞的积累。

• 发表时间: 2003
• 期刊: Brain Res 970
• 作者: Popivanova BK;Koike K;Tonchev AB;Ishida Y;Kondo T;Ogawa S;Mukaida N;Inoue M;Yamashima T.
• 通讯作者: Yamashima T.

Miyazaki, M他: "Expression of 150-kd oxygen-regulated protein in the hippocampus suppresses delayed neuronal cell death"J Cereb Blood Flow Metab.. 22. 979-987 (2002)

Miyazaki, M 等人：“海马中 150-kd 氧调节蛋白的表达抑制延迟神经元细胞死亡”J Cereb Blood Flow Metab.. 22. 979-987 (2002)