The Role of Stanniocalcin 2 in Calcium Homeostasis and Neuronal Pathology

斯钙素 2 在钙稳态和神经病理学中的作用

• 批准号: 8205022
• 负责人: William Abel Zeiger
• 金额: $ 3.69万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8205022
• 关键词: Affect; Alzheimer' s Disease; Animals; Apoptotic; Brain; Calcium; Calcium Channel; Cell membrane; Cells; Cellular Stress; Cultured Cells; Data; Disease; Disease Progression; Embryo; Exposure to; Fibroblasts; Fishes; Fluorescence Resonance Energy Transfer; Functional disorder; Genes; Glutamates; Glycoproteins; Homeostasis; Huntington Disease; Hypoxia; Injury; Ion Channel; Knockout Mice; Life; Mammalian Cell; Measures; Mediating; Mediator of activation protein; Microscopy; Middle Cerebral Artery Occlusion; Minerals; Molecular; Monitor; Nerve Degeneration; Nervous system structure; Neurodegenerative Disorders; Neuronal Injury; Neurons; Parkinson Disease; Pathogenesis; Pathology; Pathway interactions; Physiology; Play; Process; Proteins; Public Health; Rattus; Regulation; Reperfusion Therapy; Reporting; Research; Role; STIM1 gene; Site; Stroke; Testing; base; cellular imaging; design; endoplasmic reticulum stress; inhibitor/antagonist; nervous system disorder; neuron loss; neuronal survival; polyglutamine; protein function; protein misfolding; public health relevance; research study; response; sensor; stanniocalcin 2; teleocalcin

DESCRIPTION (provided by applicant): Regulation of calcium homeostasis and the unfolded protein response (UPR) are two processes of particular importance in neuronal pathophysiology. Stanniocalcin 2 (STC2) is a secreted glycoprotein thought to play a role in calcium homeostasis. Expressed as part of the UPR, STC2 is also induced in the rat brain after middle cerebral artery occlusion and knockdown of STC2 expression sensitizes cells to hypoxia and endoplasmic reticulum stress. Since STC2 is involved in both the UPR and calcium regulation, this protein may play a particularly important role in the nervous system. The objective of this proposal is to elucidate the function of STC2, with special emphasis on its possible role in neuronal pathophysiology. Aim 1: Investigate the effect of STC2 expression on STIM1 function My preliminary data demonstrates that loss of STC2 expression leads to an increase in calcium entry through store-operated calcium channels (SOCs) and that STC2 can interact with the ER calcium sensor STIM1. Therefore, I will test the hypothesis that STC2 affects calcium entry by altering STIM1 mediated regulation of SOCs. Specifically, I will conduct live cell imaging experiments to measure translocation of STIM1 after store depletion in the presence or absence of STC2 expression. I will also test the effect of STC2 expression on the interaction between ST1M1 and SOCs by measuring FRET between STIM1 and SOCs before and after calcium store depletion. Aim 2: Characterize STC2 mediated effects on calcium homeostasis following glutamate exposure STC2 is expressed in the brain following injury and plays a cytoprotective role in cultured cells. In addition, loss of STC2 expression leads to a significant increase in calcium influx. Therefore, I will test the hypothesis that STC2 mediated regulation of calcium homeostasis is important for neuronal survival in the context of calcium dependent neuronal injury. I will compare changes in [Ca2+]i levels between STC2+/+ and STC2-/- neurons following exposure to glutamate. I will also use inhibitors of store-operated calcium entry to determine the importance of this pathway in STC2 mediated effects on glutamate-induced neuronal injury.

描述（由申请人提供）：钙稳态调节和未折叠蛋白反应（UPR）是神经元病理生理学中特别重要的两个过程。斯钙素2（Stanniocalcin 2，STC 2）是一种分泌型糖蛋白，被认为在钙稳态中起作用。作为UPR的一部分表达，STC 2也在大脑中动脉闭塞后的大鼠脑中被诱导，并且STC 2表达的敲低使细胞对缺氧和内质网应激敏感。由于STC 2参与UPR和钙的调节，这种蛋白质可能在神经系统中发挥特别重要的作用。该提案的目的是阐明STC 2的功能，特别强调其在神经元病理生理学中的可能作用。目标1：研究STC 2表达对STIM 1功能的影响我的初步数据表明，STC 2表达的缺失导致通过钙库操纵的钙通道（SOC）的钙进入增加，并且STC 2可以与ER钙传感器STIM 1相互作用。因此，我将检验STC 2通过改变STIM 1介导的SOC调节来影响钙进入的假设。具体来说，我将进行活细胞成像实验，以测量STIM 1易位后，存储耗尽在存在或不存在STC 2表达。我还将通过测量STIM 1和SOC之间的FRET在钙库耗尽之前和之后测试STC 2表达对ST 1 M1和SOC之间的相互作用的影响。目标二：表征STC 2介导的谷氨酸暴露后对钙稳态的影响STC 2在损伤后的脑中表达，并在培养的细胞中起细胞保护作用。此外，STC 2表达的丧失导致钙内流的显著增加。因此，我将测试的假设，即STC 2介导的钙稳态调节是重要的钙依赖性神经元损伤的背景下，神经元的存活。我将比较暴露于谷氨酸后STC 2 +/+和STC 2-/-神经元之间[Ca 2 +]i水平的变化。我还将使用钙库操作的钙进入抑制剂，以确定在STC 2介导的谷氨酸诱导的神经元损伤的影响，这一途径的重要性。