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阳离子加合物对大鼠脊髓损伤模型基质辅助激光解吸电离成像质谱数据分析的影响。

Influence of the Cation Adducts in the Analysis of Matrix-Assisted Laser Desorption Ionization Imaging Mass Spectrometry Data from Injury Models of Rat Spinal Cord.

机构信息

Department of Physical Chemistry, Faculty of Science and Technology, University of the Basque Country (UPV/EHU) , Barrio Sarriena s/n, 48940, Leioa, Spain.

Laboratory of Molecular Neurology, Hospital Nacional de Parapléjicos (HNP) , Finca la Peraleda s/n, 45071 Toledo, Spain.

出版信息

Anal Chem. 2017 Aug 15;89(16):8565-8573. doi: 10.1021/acs.analchem.7b02650. Epub 2017 Jul 27.

Abstract

Imaging mass spectrometry (IMS) is quickly becoming a technique of reference to visualize the lipid distribution in tissue sections. Still, many questions remain open, and data analysis has to be optimized to avoid interpretation pitfalls. Here we analyze how the variation on the [Na]/[K] relative abundance affects the detection of lipids between sections of spinal cord of (uninjured) control rats and of models of spinal cord demyelination and traumatic contusion injury. The [M + Na]/[M + K] adducts ratio remained approximately constant along transversal and longitudinal sections of spinal cord from control animals, but it strongly changed depending on the type of lesion. A substantial increase in the abundance of [M + Na] adducts was observed in samples from spinal cord with demyelination, while the intensity of the [M + K] adducts was stronger in those sections from mechanically injured spinal cords. Such changes masked the modifications in the lipid profile due to the injury and only after summing the signal intensity of all adducts and corresponding monoprotonated molecular ions of each detected lipid in a single variable, it was possible to unveil the real changes in the lipid profile due to the lesion. Such lipids included glycerophospholipids (both diacyl and aryl-acyl), sphingolipids, and nonpolar lipids (diacyl and triacylglycerols), which are the main lipid classes detected in positive-ion mode. Furthermore, the results demonstrate the sensitivity of the technique toward modification in tissue homeostasis and that the [M + Na]/[M + K] ratio may be used to detect alterations in such homeostasis.

摘要

成像质谱 (IMS) 正在迅速成为一种可视化组织切片中脂质分布的参考技术。然而,仍有许多问题悬而未决,数据分析必须进行优化,以避免解释陷阱。在这里,我们分析了[Na]/[K]相对丰度的变化如何影响脊髓(未受伤)对照大鼠和脱髓鞘及创伤性挫伤损伤模型脊髓切片之间脂质的检测。[M + Na]/[M + K]加合物的比率在对照动物的脊髓横切和纵切面上基本保持不变,但它强烈地取决于病变的类型。在脱髓鞘脊髓的样本中观察到[M + Na]加合物的丰度显著增加,而在机械损伤脊髓的切片中,[M + K]加合物的强度更强。这些变化掩盖了由于损伤引起的脂质谱的变化,只有在将所有加合物的信号强度相加,并将每个检测到的脂质的相应单质子化分子离子汇总到一个单一变量中之后,才能揭示由于损伤引起的脂质谱的真正变化。这些脂质包括甘油磷脂(二酰基和芳酰基)、鞘脂和非极性脂质(二酰基和三酰基甘油),它们是正离子模式下检测到的主要脂质类。此外,结果证明了该技术对组织内稳态变化的敏感性,并且[M + Na]/[M + K]比值可用于检测这种内稳态的改变。

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