CALCIUM AND POTASSIUM CHANNELS IN LYMPHOCYTE

淋巴细胞中的钙和钾通道

• 批准号: 6564583
• 负责人: AMELIA RIVERA
• 金额: $ 14.53万
• 依托单位: UNIVERSIDAD CENTRAL DEL CARIBE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-15 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6564583
• 关键词: SDS polyacrylamide gel electrophoresis; T cell receptor; T lymphocyte; calcium channel; calcium flux; caveolins; cell line; confocal scanning microscopy; coral; diterpenes; hormone regulation /control mechanism; human tissue; immunity; immunofluorescence technique; immunomodulators; leukocyte activation /transformation; marine toxins; micromanipulator; potassium channel; progesterone; protein binding; steroid hormone; voltage /patch clamp; voltage gated channel; western blottings

The major goal of this project is to elucidate the mechanisms of non- genomic steroid effects in T lymphocytes. Non-genomic steroid actions have received much attention in the past decade. They are characterized by their rapid onset of action and by being independent of transcription and protein synthesis. Non-genomic steroid actions are ubiquitous and well documented. Steroid hormones have been shown by us and others to cause rapid changes in calcium signals in lymphocytes. These changes fall into the category of non- genomic steroid effects. In lymphocytes, initial calcium signaling upon TCR activation has been shown to occur by a two step process involving IP3 mediated release of calcium from internal stores and calcium influx through Ca2+ release-activated Ca2+ channels. This signaling cascade has been associated to tyrosine kinases in lipid rafts. Lipid rafts, or detergent insoluble microdomains, are surface membrane constituents that play important roles in signal transduction and membrane trafficking in lymphocytes and many other cells. We propose that non-genomic steroid actions are mediated by steroid association to the proteins of the lipid raft. The Specific Aims to be followed are: Specific Aim 1: Evaluate the non-genomic effects of the gonadal steroids on intracellular free Ca2+ levels in murine and human lymphoblastoid cell lines. Specific Aim 2: Determine the presence of caveolae and/or detergent insoluble microdomains (lipid rafts) in lymphoblastoid cell lines. Specific Aim 3: Correlate the presenc3e of caveolae and/or lipid rafts in l in lymphocytes with the non-genomic action of gonadal steroids on Ca2+ signaling. The results obtained in this project will contribute to our understanding of the cellular and subcellular mechanisms involved in signal transduction, non-genomic steroid action and for our understanding of the mechanisms involved in immunosuppression.

该项目的主要目标是阐明 T 淋巴细胞中非基因组类固醇效应的机制。在过去的十年中，非基因组类固醇作用受到了广泛关注。它们的特点是起效快且不依赖于转录和蛋白质合成。非基因组类固醇作用无处不在并且有据可查。我们和其他人已经证明类固醇激素会引起淋巴细胞中钙信号的快速变化。这些变化属于非基因组类固醇效应的范畴。在淋巴细胞中，TCR 激活时的初始钙信号传导已被证明是通过两步过程发生的，涉及 IP3 介导的钙从内部储存的释放以及钙通过 Ca2+ 释放激活的 Ca2+ 通道流入。这种信号级联与脂筏中的酪氨酸激酶有关。脂筏或去污剂不溶性微区是表面膜成分，在淋巴细胞和许多其他细胞的信号转导和膜运输中发挥重要作用。我们认为非基因组类固醇作用是通过类固醇与脂筏蛋白质的结合介导的。需要遵循的具体目标是： 具体目标 1：评估性腺类固醇对小鼠和人淋巴母细胞系细胞内游离 Ca2+ 水平的非基因组影响。具体目标 2：确定类淋巴母细胞系中是否存在小窝和/或去污剂不溶性微区（脂筏）。具体目标 3：将淋巴细胞中小凹和/或脂筏的存在与性腺类固醇对 Ca2+ 信号传导的非基因组作用相关联。该项目获得的结果将有助于我们了解信号转导、非基因组类固醇作用所涉及的细胞和亚细胞机制，以及免疫抑制所涉及的机制。