COPPER HIERARCHY IN ZEBRAFISH

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 痕量金属离子在生物学中起着关键作用。了解微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。在本提案中，我们将利用XRF成像来研究金属稳态。使用基因突变和铜螯合剂设计了铜缺乏水平增加的斑马鱼胚胎。 随着铜水平的降低，非必需的铜依赖过程，如色素沉着，丢失。 然后脊索发育受到影响，最后神经变形。 已经确定了铜层级，其中铜首先针对更重要的系统，然后针对非重要的系统。 目前还不清楚这种层次结构是在系统水平还是细胞水平上实施的，铜是否靶向负责神经发育的细胞而不是负责色素沉着的细胞，或者铜在所有细胞中的递送是否相等，但是阈值细胞铜水平需要表达负责色素沉着的蛋白质，高于表达更必需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，以绘制这种金属的系统范围贩运。

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.