Cellular Neurobiology, NIDA IRP, NIH, Baltimore, MD 21224, USA.
Glycobiology. 2010 Jun;20(6):661-7. doi: 10.1093/glycob/cwq031. Epub 2010 Feb 28.
In this study, we describe a simple and efficient method for mapping the distribution and localization of all sialylated sphingoglycolipids present in coronal mouse brain sections using a conventional axial matrix-assisted laser desorption/ionization time of flight. A single scan of a histological tissue section gives a complete profile of ganglioside species without derivatization or labeling. We have developed and tested a new matrix preparation (2,6-dihydroxyacetophenone [DHA]/ammonium sulfate/heptafluorobutyric acid [HFBA]) to maximize the detection of all ganglioside species; the ammonium sulfate limits the formation of salt adducts, while the addition of HFBA increases the stability of DHA in a vacuum, thus facilitating imaging applications. Our results, in both extracted samples and whole tissue sections using negative ion reflectron and linear modes, show differences in localization in several brain regions depending on the sialic acids and the ceramide-associated core gangliosides.
在这项研究中,我们描述了一种简单有效的方法,使用常规的轴向基质辅助激光解吸/电离飞行时间来绘制冠状小鼠脑切片中所有唾液酸化神经糖脂的分布和定位。对组织切片的单次扫描即可提供未经衍生化或标记的神经节苷脂种类的完整图谱。我们已经开发并测试了一种新的基质制备方法(2,6-二羟基苯乙酮[DHA]/硫酸铵/全氟丁酸[HFBA]),以最大限度地检测所有神经节苷脂种类;硫酸铵限制盐加合物的形成,而添加 HFBA 可增加 DHA 在真空中的稳定性,从而有利于成像应用。我们的结果表明,在使用负离子反射和线性模式的提取样本和整个组织切片中,根据唾液酸和与神经酰胺相关的核心神经节苷脂的不同,几个脑区的定位存在差异。