Dufresne Martin, Guneysu Daniel, Patterson Nathan Heath, Marcinkiewicz Mieczyslaw Martin, Regina Anthony, Demeule Michel, Chaurand Pierre
Department of Chemistry, Université de Montréal, C.P. 6128, succursale Centre-ville, Montréal, QC, H3C 3J7, Canada.
Département de chimie, Université P. et M. Curie, 4 place Jussieu, 75252, Paris, France.
Anal Bioanal Chem. 2017 Feb;409(5):1425-1433. doi: 10.1007/s00216-016-0076-x. Epub 2016 Nov 21.
Mucopolysaccharidosis type II (Hunter's disease) mouse model (IdS-KO) was investigated by both imaging mass spectrometry (IMS) and immunohistochemistry (IHC) performed on the same tissue sections. For this purpose, IdS-KO mice brain sections were coated with sublimated 1,5-diaminonaphtalene and analyzed by high spatial resolution IMS (5 μm) and anti-GM3 IHC on the same tissue sections to characterize the ganglioside monosialated ganglioside (GM) deposits found in Hunter's disease. IMS analysis have found that two species of GM3 and GM2 that are only different due to the length of their fatty acid residue (stearic or arachidic residue) were overexpressed in the IdS-KO mice compared to a control mouse. GM3 and GM2 were characterized by on-tissue exact mass and MS/MS compared to a GM3 standard. Realignment of both IMS and IHC data sets further confirmed the observed regioselective signal previously detected by providing direct correlation of the IMS image for the two GM3 overly expressed MS signals with the anti-GM3 IHC image. Furthermore, these regioselective GM MS signals were also found to have highly heterogeneous distributions within the GM3-IHC staining. Some deposits showed high content in GM3 and GM2 stearic species (r = 0.74) and others had more abundant GM3 and GM2 arachidic species (r = 0.76). Same-section analysis of Hunter's disease mouse model by both high spatial resolution IMS and IHC provides a more in-depth analysis of the composition of the GM aggregates while providing spatial distribution of the observed molecular species. Graphical Abstract Ganglioside imaging mass spectrometry followed by immunohistochemistry performed on the same tissue section.
通过对同一组织切片进行成像质谱（IMS）和免疫组织化学（IHC），对II型黏多糖贮积症（亨特氏病）小鼠模型（IdS-KO）进行了研究。为此，将IdS-KO小鼠脑切片用升华的1,5-二氨基萘包被，并通过高空间分辨率IMS（5μm）和同一组织切片上的抗GM3 IHC进行分析，以表征在亨特氏病中发现的神经节苷脂单唾液酸神经节苷脂（GM）沉积物。IMS分析发现，与对照小鼠相比，IdS-KO小鼠中两种仅因脂肪酸残基长度（硬脂酸或花生酸残基）不同的GM3和GM2物种过表达。与GM3标准品相比，通过组织上的精确质量和MS/MS对GM3和GM2进行了表征。IMS和IHC数据集的重新比对通过将两个GM3过表达MS信号的IMS图像与抗GM3 IHC图像直接关联，进一步证实了先前检测到的区域选择性信号。此外，还发现这些区域选择性GM MS信号在GM3-IHC染色内具有高度异质的分布。一些沉积物中GM3和GM2硬脂酸物种含量高（r = 0.74），而其他沉积物中GM3和GM2花生酸物种含量更丰富（r = 0.76）。通过高空间分辨率IMS和IHC对亨特氏病小鼠模型进行同一切片分析，在提供观察到的分子物种空间分布的同时，对GM聚集体的组成进行了更深入的分析。图形摘要 在同一组织切片上进行神经节苷脂成像质谱，然后进行免疫组织化学。
