Manganese-iron interactions determining neuronal function and phenotype

锰和铁是必不可少的微量金属，对大脑生理和发育至关重要。任何一种金属的失衡都会导致有害的神经发育和神经退行性疾病。[1,2]在中枢神经系统中，锰和铁有许多共同的转运蛋白，这就解释了为什么一种金属的失衡会影响另一种金属。SLC转运蛋白包括SLC30A10， SLC39A8和SLC39A14促进细胞锰和铁的摄取/输出本项目将探讨锰-铁相互作用对神经元功能的贡献，特别是锰代谢的扰动如何调节铁代谢，从而改变神经元的功能和表型。目的1：确定锰-铁相互作用对神经元功能和行为的影响。将在野生型和SLC转运体突变斑马鱼中进行全脑活动、单细胞分辨率解剖和神经化学制图以及体内钙成像，以识别依赖严格金属稳态的神经元回路。我们将特别关注视觉通路，因为斑马鱼的视网膜对金属扰动特别敏感视觉和运动行为分析将功能/分子变化与行为改变联系起来。CRISPR/Cas9将用于确定分子事件的层次结构。目标2：确定锰-铁相互作用对组织和细胞区室金属分布的影响，并将空间变化与功能联系起来。该学生将参加伦敦金属学设施，使用激光烧蚀电感耦合等离子体质谱（LA-ICP-MS）生成斑马鱼大脑和眼睛的定量“金属图”（Mn,Fe, Zn, Cu）。荧光铁探针将用于评估斑马鱼神经元培养中细胞间的相对铁分布，调节功能和行为表型。Tuschl博士是锰神经科学[1,3,4]方面的专家，他将在这个项目中领导斑马鱼成像和行为分析。苏博士的研究重点是铁在脑老化和神经变性中的作用，他将在铁稳态和定量金属分析/成像方面提供专业知识。[2,5]

Manganese and iron are essential trace metals and critical for brain physiology anddevelopment. Dyshomeostasis of either metal leads to detrimental neurodevelopmental andneurodegenerative disorders.[1,2] Numerous transporters in the central nervous system areshared between manganese and iron, explaining why dyshomeostasis of one metal impactsthe other. SLC transporters including SLC30A10, SLC39A8 and SLC39A14 facilitate bothcellular manganese and iron uptake/export.[3] This project will explore the contribution ofmanganese-iron interactions to neuronal function, especially how perturbation of manganesemetabolism can modulate iron metabolism and together, alter neuronal function andphenotype.Objective 1: Determine the effect of manganese-iron interactions on neuronal function andbehaviour.Whole brain activity, anatomy and neurochemistry mapping at single-cell resolution as wellas in vivo calcium imaging will be performed in wild-type and SLC transporter mutantzebrafish to identify neuronal circuits dependent on strict metal homeostasis. There will be aspecial focus on the visual pathway as the zebrafish retina is particularly sensitive to metalperturbation.4 Visual and locomotor behavioural analysis will link functional/molecularchanges to altered behaviour. CRISPR/Cas9 will be used to determine the hierarchy ofmolecular events.Objective 2: Determine the effect of manganese-iron interactions on tissue and cellularcompartmental metal distributions and linking spatial changes to function under item 1.The student will attend the London Metallomics Facility to use laser ablation inductivelycoupled-plasma mass spectrometry (LA-ICP-MS) to generate quantitative "metal maps" (Mn,Fe, Zn, Cu) of the zebrafish brain and eye. Fluorescent iron probes will be used to assessrelative iron distributions between cellular compartments in zebrafish neuronal cultures,modulating function and behavioural phenotype.Dr Tuschl, expert in manganese neuroscience [1,3,4], will lead on the zebrafish imaging andbehavioural analysis in this project. Dr So, whose research focuses on the role of iron on brainageing and neurodegeneration, will provide expertise in iron homeostasis and quantitativemetal analysis/imaging.[2,5]