PROCESSES OF METAL SELECTION BY METAL ACCUMULATING CELLS

金属积累池的金属选择过程

• 批准号: 3438158
• 负责人: ANDREW Z MASON
• 金额: $ 6.66万
• 依托单位: CALIFORNIA STATE UNIVERSITY LONG BEACH
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-05-01 至 1991-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3438158
• 关键词: Gastropoda; basophils; cell membrane; cell type; chemical bond; copper; eukaryote; gel electrophoresis; high performance liquid chromatography; ion transport; mass spectrometry; metal metabolism; phagocytosis; pinocytosis; receptor mediated endocytosis; schistosomiasis; tissue /cell culture

The purpose of this project is to identify and characterize the mechanisms which enable the pore cells of the marine prosobranch gastropod Littorina littorea to specifically accumulate Cu in preference to other bioavailable metals. A series of experiments have been designed to demonstrate that these cells accumulate metals from the blood and to test if the observed selectivity shown by the cells in vivo is due to: (1) intrinsic properties unique to this particular cell type; (2) the activities of other cell types such as the basophil cells in the removal of metals other than Cu from the blood; (3) humoral factors which bind to Cu and act to both vector the metal to the cell and facilitate its uptake. A number of techniques including cell culture, energy dispersive X-ray microanalysis, radiometric analysis, gel electrophoresis, size exclusion high performance liquid chromatography and inductively coupled plasma mass spectrometry will be used to resolve the relative importance of the three factors outlined above. Short term kinetic studies will also be undertaken to determine if Cu uptake by the cells is carrier mediated. Competitive inhibition studies with other metals will be used to determine the specificity of the uptake system and the energetic requirements of the transport system will also be studied using various metabolic inhibitors. In combination, these studies will provide a mechanistic understanding of the processes which permit the sequential recognition, selection and vectoring of metals to specific cell types in higher eukaryotes. In the long term, these studies will also help in the development of Cu based molluscicides to control the spread of schistosomiasis by the gastropod Biomphalaria glabrata. Finally, these studies represent the first attempt to study the flux and turnover of Cu in cells quantitatively using stable isotopic analysis of 63Cu by inductively coupled plasma mass spectrometry. This technique therefore offers a new methodology to the health related sciences for the study of this essential, but potentially toxic element.

该项目的目的是识别和表征 使海洋孔单元的机制 Prosobranch腹足动物Littorina Littorea专门 优先考虑其他生物利用金属。 一个 已经设计了一系列实验来证明 这些细胞从血液中积聚金属，以测试是否存在 观察到的体内细胞显示的选择性是由于：（1） 这种特定细胞类型独有的内在属性； （2） 其他细胞类型的活动，例如嗜碱性粒细胞 从血液中除去除铜以外的金属； （3）体液 与Cu结合并作用于金属将金属载入的因素 细胞并促进其吸收。 许多技术，包括 细胞培养，能量色散X射线微分析，辐射计 分析，凝胶电泳，尺寸排除高性能 液相色谱和电感耦合等离子体质量 光谱法将用于解决相对重要性 上面概述的三个因素。 短期动力学研究将 还要确定细胞摄取的Cu是否为 载体介导。 与其他的竞争性抑制研究 金属将用于确定摄取的特异性 运输系统的系统和充满活力的要求 还将使用各种代谢抑制剂研究。 在 结合这些研究将提供机械性 了解允许顺序的过程 将金属识别，选择和向矢量矢量对特定细胞 较高的真核生物类型。 从长远来看，这些研究将 也有助于开发基于Cu的软蛋白酶来控制 血吸虫病生物脑血吸虫病的传播 Glabrata。 最后，这些研究是首次尝试 研究细胞中CU的通量和转移，并使用 通过电感耦合等离子体对63CU的稳定同位素分析 质谱法。 因此，该技术提供了新的 研究与健康有关的科学的方法论 必不可少但可能有毒的元素。