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: 8011946
• 负责人: William Abel Zeiger
• 金额: $ 3.52万
• 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/8011946
• 关键词: Affect; Alzheimer' s Disease; Animals; Apoptotic; Brain; Calcium; Calcium Channel; Cell membrane; Cells; Cellular Stress; Critiques; Cultured Cells; Data; Disease; Disease Progression; Embryo; Exposure to; Fibroblasts; Fishes; Fluorescence Resonance Energy Transfer; Functional disorder; Genes; Glutamates; Glycoproteins; Homeostasis; Huntington Disease; Hypoxia; Individual; 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; Outcome; Parkinson Disease; Pathogenesis; Pathology; Pathway interactions; Physiology; Play; Process; Proteins; Public Health; Rattus; Regulation; Reperfusion Therapy; Reporting; Research; Role; STIM1 gene; Site; Stroke; Testing; Update; base; cellular imaging; design; endoplasmic reticulum stress; inhibitor/antagonist; meetings; 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. Public Health Relevance: Neurological disorders pose a significant public health burden. STC 2 may play an important role in fundamental processes of the nervous system. Therefore, these studies will increase our understanding of the basic mechanisms underlying neuronal injury and neurodegenerative disease. NOTE: The critiques of individual reviewers are provided below in an essentially unedited form. These critiques were prepared prior to the review meeting and may not have been updated or revised subsequent to the discussion at the meeting. Therefore, they may not fully reflect the final opinions of the individual reviewers at the close of group discussion or the final majority opinion of the group. The Resume and Summary of Discussion above summarizes the final outcome of the group discussion.

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。斯坦钙素2 （STC2）是一种分泌糖蛋白，被认为在钙稳态中起作用。STC2作为UPR的一部分表达，在大脑中动脉闭塞后，STC2也在大鼠大脑中被诱导表达，抑制STC2表达使细胞对缺氧和内质网应激敏感。由于STC2参与UPR和钙的调节，该蛋白可能在神经系统中发挥特别重要的作用。本文的目的是阐明STC2的功能，特别强调其在神经元病理生理中的可能作用。目的1：研究STC2表达对STIM1功能的影响我的初步数据表明，STC2表达的缺失导致钙通过储存操作钙通道（soc）进入增加，STC2可以与内质网钙传感器STIM1相互作用。因此，我将检验STC2通过改变STIM1介导的SOCs调节来影响钙进入的假设。具体而言，我将进行活细胞成像实验，以测量STC2表达存在或不存在的情况下，存储耗尽后STIM1的易位。我还将通过测量钙储存耗尽前后STIM1与soc之间的FRET来测试STC2表达对ST1M1与soc之间相互作用的影响。目的2：表征STC2对谷氨酸暴露后钙稳态的影响STC2在脑损伤后表达，并在培养细胞中发挥细胞保护作用。此外，STC2表达缺失导致钙内流显著增加。因此，我将验证STC2介导的钙稳态调节对钙依赖性神经元损伤背景下神经元存活的重要假设。我将比较STC2+/+和STC2-/-神经元在暴露于谷氨酸后[Ca2+] I水平的变化。我还将使用储存操作钙进入抑制剂来确定该途径在STC2介导的谷氨酸诱导的神经元损伤中的重要性。