THE ROLE OF CALCIUM IN THE CYTOTOXCICTY OF CHLOROPHENOLS

钙在氯酚细胞毒性中的作用

• 批准号: 3755890
• 负责人: JUDE C NWOGA
• 金额: --
• 依托单位: FLORIDA AGRICULTURAL AND MECHANICAL UNIV
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3755890
• 关键词: Cirripedia; alternatives to animals in research; biological signal transduction; calcium channel blockers; calcium flux; calcium indicator; calmodulin; cell membrane; cytotoxicity; fluorescence spectrometry; membrane proteins; muscle cells; phenols; phosphorylation

The goal of this proposal is to determine the membrane mechanism of action of chlorophenols. In this project, the role of calcium in the cytotoxicity of chlorophenols in excitable tissues, will be investigated. Single muscle fibers from the giant barnacle, will be used as a model preparation for these studies. Specific objectives of the project are: 1. To determine if exposure of barnacle muscle fibers to chlorophenols induce an increase in intracellular calcium levels in a dose and time dependent manner and whether the effect can be blocked by calcium channel antagonists. 2. To determine if the induced calcium accumulation activates the calcium messenger system, e. calcium/calmodulin protein kinase II. 3. To determine if the activated calcium messenger system results in the phosphorylation of specific membrane proteins, e.g., Na/Ca exchanger. 4. To train students in the techniques of working with single muscle cells to investigate the membrane mechanism of action of cytotoxic agents. Objective 1 will be carried out by 45Ca2+ uptake and Fura-2 fluorescence studies in the absence and presence of calcium channel antagonists. Objective 2 and 3 will be carried out by phosphorylation studies in the absence and presence of calmodulin inhibitors. Objective 4 will be carried out by recruiting two students under the MBRS program for participation in hands on research. Results obtained from using this preparation, will help provide insight to the physiological parameters modulated, and the consequent responses that are initiated early in the presence of chlorophenols, which may lead eventually to cell damage and death, especially in the excitable tissues. This may have an impact on mitigation of toxicity of chlorophenols.

这项建议的目标是确定膜机制 氯代酚的作用。在这个项目中，钙在体内的作用 氯酚在可兴奋组织中的细胞毒性将被研究。 来自巨型藤壶的单一肌肉纤维将被用作模型 为这些研究做准备。该项目的具体目标是： 1.确定氯苯酚对藤壶肌纤维的暴露 在一定剂量和时间内诱导细胞内钙水平增加 依赖方式及钙通道能否阻断其作用 对抗者。2.确定诱导的钙蓄积是否 激活钙信使系统，即钙/钙调蛋白 3.确定激活的钙信使系统是否 导致特定膜蛋白的磷酸化，例如Na/Ca 交易所。4.培训学生与单身人士合作的技巧 肌细胞膜细胞毒作用机制的研究 探员们。目标1将通过45Ca+摄取和Fura-2来实现 钙通道存在和不存在时的荧光研究 对抗者。目标2和目标3将通过磷酸化实现 钙调蛋白抑制剂的存在和缺失的研究。客观化 4将通过在MBRS计划下招收两名学生来实施 参与亲身实践研究。通过使用此工具获得的结果 准备，将有助于深入了解生理参数 调制的，以及在早期启动的后续响应 氯酚的存在，最终可能导致细胞损伤和 死亡，尤其是在可兴奋的组织中。这可能会对 降低氯代酚的毒性。