TOMOGRAPHIC DETERMINATION OF ZEBRAFISH METAL DISTRIBUTIONS

这个子项目是许多研究子项目中的一个 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。 铜是一种重要的过渡金属，在需氧生物的生化中起着关键作用。 Gitlin及其同事开发了一种化学遗传筛选方法，用于识别选择性干扰斑马鱼铜稳态的小分子。 这使得几种分子，可用于选择性地改变铜浓度在斑马鱼胚胎发育，从而提供深入了解铜代谢的发育病理生理学，建议的方式，其中次优的铜代谢可能有助于出生缺陷。 例如，他们发现了铜代谢的适应性层次，治疗时间不同和/或抑制剂浓度不同，首先影响黑色素沉着，然后影响心室大小，然后影响脊索发育。 这种行为重现了在人类门克斯病患者中观察到的病理学。 我们建议使用X-射线微探针成像，以确定所有的必需微量元素（V，Mn，Fe，Co，Ni，Cu和Zn）在斑马鱼胚胎中的分布和浓度的选择性铜depelection在不同的时间在发展过程中的功能。 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组复杂的相互作用，因此探索不同必需微量元素在不同Cu胁迫条件下的完整分布是重要的。

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.