X-ray fluorescence analysis of iron and manganese distribution in primary dopaminergic neurons.
X-ray fluorescence analysis of iron and manganese distribution in primary dopaminergic neurons.
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DOI：
10.1111/jnc.12073
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发表时间：
2013-01
影响因子：
4.7
通讯作者：
Lingor P
中科院分区：
文献类型：
--
作者：
Dučić T;Barski E;Salome M;Koch JC;Bähr M;Lingor P
过渡金属已被认为在帕金森病的发病机制中起关键作用。x射线显微镜结合低温设置是一个强大的方法，在低浓度和高分辨率的完整细胞元素成像，消除了固定和标本切片的需要。在这里，我们使用x射线荧光显微镜对培养的初级中脑神经元进行了元素分布分析，步长为300 nm，灵敏度为0.1 ppm，低温条件下。我们报告了过渡金属处理后小鼠多巴胺能神经元和非多巴能神经元在亚细胞水平上的元素映射。Fe2+的应用导致铁在细胞外大量积累，而对神经元递质亚型没有偏好。利用x射线吸收近边结构分析对不同的铁氧化态进行了定量分析。Mn2+处理后，能神经元中Mn的胞质/核旁定位优先，而Mn3+处理后未检测到明显的信号。免疫细胞化学分析表明，能神经元对锰的优先摄取与电压门控钙通道的表达增加有关。我们讨论了这种差异元素分布对能神经元选择性易感性和帕金森病发病机制的影响。
Transition metals have been suggested to play a pivotal role in the pathogenesis of Parkinson's disease. X-ray microscopy combined with a cryogenic setup is a powerful method for elemental imaging in low concentrations and high resolution in intact cells, eliminating the need for fixation and sectioning of the specimen. Here, we performed an elemental distribution analysis in cultured primary midbrain neurons with a step size in the order of 300 nm and ∼ 0.1 ppm sensitivity under cryo conditions by using X-ray fluorescence microscopy. We report the elemental mappings on the subcellular level in primary mouse dopaminergic (DAergic) and non-DAergic neurons after treatment with transition metals. Application of Fe2+ resulted in largely extracellular accumulation of iron without preference for the neuronal transmitter subtype. A quantification of different Fe oxidation states was performed using X-ray absorption near edge structure analysis. After treatment with Mn2+, a cytoplasmic/paranuclear localization of Mn was observed preferentially in DAergic neurons, while no prominent signal was detectable after Mn3+ treatment. Immunocytochemical analysis correlated the preferential Mn uptake to increased expression of voltage-gated calcium channels in DAergic neurons. We discuss the implications of this differential elemental distribution for the selective vulnerability of DAergic neurons and Parkinson's disease pathogenesis.
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影响因子：
4.3
作者：
Jomova, Klaudia;Vondrakova, Dagmar;Valko, Marian
通讯作者：
Valko, Marian
影响因子：
3.4
作者：
Crossgrove, JS;Yokel, RA
通讯作者：
Yokel, RA
影响因子：
--
作者：
Edwards, Jonathan
通讯作者：
Edwards, Jonathan
影响因子：
5.3
作者：
Lingor, P;Unsicker, K;Krieglstein, K
通讯作者：
Krieglstein, K
影响因子：
4.5
作者：
Benedetto, Alexandre;Au, Catherine;Aschner, Michael
通讯作者：
Aschner, Michael