TOMOGRAPHIC DETERMINATION OF ZEBRAFISH METAL DISTRIBUTIONS

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。 铜是一种重要的过渡金属，在好氧生物的生物化学中起着至关重要的作用。吉特林和他的同事(提交)已经开发出一种化学基因筛查，用于识别选择性地扰乱斑马鱼体内铜稳态的小分子。这使得可以识别出几个分子，可以用来选择性地改变斑马鱼胚胎发育中的铜浓度，从而为铜代谢的发育病理生理学提供了洞察，提出了次优铜代谢可能导致出生缺陷的途径。例如，他们发现了铜代谢的一个适应性等级，用不同的时间长度和/或不同浓度的抑制剂进行治疗，首先影响黑色素的沉积，然后是脑室大小，然后是脊索发育。这种行为重现了在人类门克斯病患者身上观察到的病理。我们建议使用X射线微探针成像技术来确定所有必需的微量元素(V、Mn、Fe、Co、Ni、铜和锌)在斑马鱼胚胎中的分布和浓度，作为斑马鱼胚胎发育过程中不同时期铜选择性降解的函数。以前的研究(例如，在酵母模型中)揭示了不同必需金属浓度之间的一系列复杂的相互作用，因此探索不同铜胁迫条件下不同必需微量元素的完整分布是很重要的。

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. Copper is an essential transition metal that plays a critical role in the biochemsitry of aerobic organisms. Gitlin and co-workers (submitted) have developed a chemical genetic screen for identifying small molecules that selectively perturb copper homeostasis in zebrafish. This has allowed the identification of several molecules that can be used to selectively alter Cu concentration in developing zebrafish embryos, thus providing insight into the developmental pathophysiology of copper metabolism, suggesting ways in which suboptimal copper metabolism may contribute to birth defects. For example, they have found an adaptive hierachy of copper metabolism, with treatments for different lengths of time and/or with different concentration of inhibitor, affecting first melanin pigmentation, then ventricle size, then notochord development. This behavior reproduces the pathology observed in human Menkes-disease patients. We propose to use x-ray microprobe imaging to determine the distribution and concentration of all of the essential trace elements (V, Mn, Fe, Co, Ni, Cu, and Zn) in zebrafish embryos as a function of selective Cu depeletion at various times during development. Previous studies (e.g., in yeast models) have revealed a complex set of interactions between the concentrations of different essential metals, and thus it is important to explore the full distribution of different essential trace elements under different conditions of Cu stress.