COPPER HIERARCHY IN ZEBRAFISH

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 痕量金属离子在生物学中起着至关重要的作用。了解痕量金属的无机生理学的一个关键方面是检测元素在组织中的分布。在这项提议中，我们将利用XRF成像来研究金属的动态平衡。利用基因突变和铜络合剂设计了铜缺乏程度越来越高的斑马鱼胚胎。随着铜水平的下降，非必需的铜依赖过程，如色素沉着，就会消失。然后脊索发育受到影响，最终导致神经变形。已经确定了铜层次结构，其中铜首先针对更基本的系统，然后针对非基本系统。目前尚不清楚这种分级是在系统水平还是细胞水平上实施的，铜是针对负责神经发育的细胞而不是负责色素沉着的细胞，还是铜在所有细胞中的输送是相同的，但细胞内铜的阈值水平需要表达负责色素形成的蛋白质高于表达更重要的蛋白质所需的蛋白质。XRF成像将用于研究铜在斑马鱼胚胎中的分布，以绘制这种金属在整个系统中的运输情况。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Trace metal ions play a critical role in biology. One critical aspect of understanding the inorganic physiology of trace metals is detection of the distribution of the elements in tissues. In this proposal we will be utilising XRF imaging to study the metal homeostasis. Zebrafish embryos with increasing levels of copper deficiency have been designed using genetic mutation and copper chelators. As copper levels decrease, non-essential copper dependent processes, such as pigmentation, are lost. Then notochord development is affected and finally neurological deformation. A copper hierarchy has been identified where copper is target to more essential systems first and then to non-essential systems. It is unknown whether this hierarchy is enforced on a systemic level or a cellular level, is copper targeted to the cells responsible for neurological development over those responsible for pigmentation, or is copper delivery equal in all cells but the threshold cellular copper levels need to express the proteins responsible for pigmentation higher that those needed for expression of more essential proteins. XRF imaging will be used to study the distribution of copper in Zebrafish embryos to map the system wide trafficking of this metal.