Manganese-iron interactions determining neuronal function and phenotype

锰和铁是必需的微量金属，对大脑的生理和发育至关重要。任何一种金属的稳态异常都会导致有害的神经发育和神经退行性疾病。[1，2]中枢神经系统中的许多转运蛋白在锰和铁之间共享，这解释了为什么一种金属的稳态失调会影响另一种金属。SLC转运蛋白SLC 30 A10、SLC 39 A8和SLC 39 A14促进细胞锰和铁的摄取/输出。[3]本项目将探讨锰-铁相互作用对神经元功能的影响，特别是锰-铁相互作用的干扰如何调节铁代谢，并共同改变神经元的功能和表型。确定锰-铁相互作用对神经元功能和行为的影响。解剖和神经化学映射在单细胞分辨率以及在体内钙成像将在野生型，型和SLC转运蛋白muscle斑马鱼，以确定依赖于严格的金属稳态的神经元电路。由于斑马鱼视网膜对金属扰动特别敏感，因此将特别关注视觉通路。4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。确定锰-铁相互作用对组织和细胞间室金属分布的影响，并将空间变化与第1项下的功能联系起来。学生将参加伦敦金属组学设施，使用激光消融感应耦合等离子体质谱法利用LA-ICP-MS技术，生成斑马鱼脑和眼的定量“金属图谱”（Mn、Fe、Zn、Cu）。荧光铁探针将用于评估斑马鱼神经元培养物中细胞区室之间的相对铁分布，调节功能和行为表型。Tuschl博士是锰神经科学专家[1，3，4]，将在该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退化中的作用，他将提供铁稳态和定量金属分析/成像方面的专业知识。[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]