THE ROLE OF CALCIUM IN THE CYTOTOXCICTY OF CHLOROPHENOLS

钙在氯酚细胞毒性中的作用

• 批准号: 5211671
• 负责人: JUDE C NWOGA
• 金额: --
• 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/5211671
• 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.

这项提议的目的是确定膜机制， 氯酚的作用。 在这个项目中，钙在 将研究氯酚在可兴奋组织中的细胞毒性。 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.为了确定藤壶肌肉纤维暴露于氯酚 在一定剂量和时间内诱导细胞内钙水平增加 依赖性方式及该作用是否可被钙通道阻断 对手。 2.为了确定诱导的钙积累是否 激活钙信使系统，e.钙/钙调蛋白 激酶II。 3.为了确定激活的钙信使系统 导致特异性膜蛋白的磷酸化，例如，Na/CA 交换器 4.训练学生在工作中的技术与单一的 肌细胞研究细胞毒作用的膜机制 剂. 目的1将通过45 Ca 2+摄取和Fura-2来进行 在钙通道存在和不存在的情况下的荧光研究 对手。 目标2和3将通过磷酸化进行 在不存在和存在钙调素抑制剂的情况下的研究。 目的 4将通过在MBRS计划下招募两名学生进行 参与研究的机会。 使用此方法获得的结果 准备，将有助于提供洞察生理参数 调制，以及随后的反应，在早期启动的 氯酚的存在，这可能最终导致细胞损伤， 死亡，特别是在易兴奋的组织中。 这可能会影响到 减轻氯酚的毒性。