Control of Ca signaling and apoptosis by InsP3 receptors

InsP3 受体对 Ca 信号传导和细胞凋亡的控制

• 批准号: 6951904
• 负责人: MICHAEL H NATHANSON
• 金额: $ 4.03万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6951904
• 关键词: Brazil; acinar cell; apoptosis; basolateral membrane; biological signal transduction; calcium channel; calcium flux; calcium ion; calcium metabolism; cell growth regulation; confocal scanning microscopy; cooperative study; epithelium; exocytosis; flash photolysis; gap junctions; liver cells; liver function; pancreas; protein localization; receptor expression; second messengers; secretion

DESCRIPTION (provided by applicant) Cytosolic Ca 2+ regulates a wide range of cell functions, from secretion to metabolism to cell growth and death. It is unknown how Ca 2+ simultaneously controls such diverse activities in an individual cell, although expression of multiple isoforms of the inositol 1,4,5-trisphosphate (InsP3) receptor (InsP3R)/Ca 2+ release channel may be responsible by allowing distinct Ca 2+ signals to occur in different subcellular regions. The parent grant for this FIRCA proposal tests the hypothesis that Ca 2+ waves in hepatocytes result from the coordinated release of Ca 2+ from distinct InsP3Rs, and that localized, subcellular increases in Ca 2+ can regulate secretion. The current proposal would complement the hypothesis of the parent grant in an important way, by testing whether distinct isoforms of the InsP3R regulate apoptosis. This question is of fundamental importance in cell biology, since apoptosis contributes to the regulation of cell growth and death in virtually all tissues, and since the InsP3R may be necessary to mediate apoptosis. The specific aims of this project are: 1. To characterize the Ca 2+ signaling machinery in a model epithelial cell line that expresses all three isoforms of the InsP3R. We will examine the expression and subcellular distribution of InsP3Rs and the pattern of InsP3-induced Ca 2+ signals in CHO cells, a model epithelial cell line that expresses all three InsP3R isoforms. 2. To investigate the contribution of each isoform of the InsP3R to the pattern of Ca 2+ signaling. We will determine the contribution of each InsP3R isoform to Ca 2+signaling in CHO cells by selectively inhibiting expression of different combinations of InsP3R isoforms while Ca 2+ signals are monitored by confocal microscopy. 3. To determine the effect of each isoform of the InsP3R on apoptosis. We will examine bile acid-induced apoptosis in CHO cells expressing the bile acid uptake protein NTCP plus different combinations of InsP3R isoforms. This research will be performed primarily at UFMG in Brazil in collaboration with M. Fatima Leite as an extension of NIH R01 DK45710.

描述（由申请人提供） 胞质Ca 2+调节从分泌到新陈代谢到细胞生长和死亡的广泛细胞功能。尚不清楚Ca 2+如何同时控制单个细胞中的这种不同活性，尽管肌醇1,4,5-三磷酸（INSP3）受体（INSP3R）/Ca 2+释放通道的多种同工型的表达可以通过允许不同的Ca 2+信号来造成不同的亚c纤维下区域。该FIRCA提案的父赠款检验了以下假设：肝细胞中的Ca 2+波是由从不同的INSP3中协调释放的Ca 2+引起的，并且Ca 2+的局部亚细胞增加可以调节分泌。当前的提议将通过测试INSP3R的不同同工型是否调节细胞凋亡的不同同工型来补充父授予的假设。这个问题在细胞生物学中至关重要，因为细胞凋亡几乎有助于调节细胞生长和死亡，并且由于INSP3R可能是介导凋亡所必需的。该项目的具体目的是： 1。在模型上皮细胞系中表征Ca 2+信号机械，该细胞系表示INSP3R的所有三种同工型。我们将检查INSP3R的表达和亚细胞分布以及CHO细胞中INSP3诱导的Ca 2+信号的模式，Cho细胞是一种模型上皮细胞系，表达了所有三个INSP3R同工型。 2。研究INSP3R对Ca 2+信号传导模式的每个同工型的贡献。我们将通过选择性地抑制INSP3R同工型的不同组合的表达，而Ca 2+信号通过共聚焦显微镜监测，我们将通过选择性抑制不同组合的表达来确定CHO细胞中CA 2+信号传导的贡献。 3。确定INSP3R每种同工型对细胞凋亡的影响。我们将检查表达胆汁酸摄取蛋白NTCP的CHO细胞中胆汁酸诱导的凋亡，以及INSP3R同工型的不同组合。 这项研究将主要在巴西的UFMG与Fatima Leite合作，作为NIH R01 DK45710的扩展。