Analysis Division, Kaneka Techno Research Corporation, 1-8 Miyamae-cho Takasago-cho, Takasago City, Hyogo, 676-8688, Japan.
Graduate School of Advanced Sciences of Matter, Hiroshima University, 1-3-1 Kagamiyama Higashihiroshima, Hiroshima, 739-8530, Japan.
Glycoconj J. 2021 Apr;38(2):167-175. doi: 10.1007/s10719-021-09984-w. Epub 2021 Mar 12.
Glycans play an important physiological role and are drawing attention as biomarkers that capture pathophysiological changes. Glycans can be detected by mass spectrometry, but recently matrix-assisted laser desorption/ionization- mass spectrometry imaging (MALDI-MSI) has enabled the visualization of glycans distribution on tissues. In this study, focusing on sialylated glycan (sialoglycans), we investigated the amidation reaction used to visualize glycans distribution, and developed a method of sialic acid derivatization by benzylamidation which is more sensitive than conventional amidation. Furthermore, we adapted this method for visualizing glycans in formalin-fixed paraffin-embedded (FFPE) liver tissue from normal mice and non-alcoholic steatohepatitis (NASH) model mice using MALDI-MSI. As a result, an increase in the distribution of glycan N-Acetylneuraminic acid-(NeuAc) ions was observed in the NASH mouse liver, and the change in glycan structure in the NASH model was clarified.
聚糖具有重要的生理作用,并作为捕获病理生理变化的生物标志物引起了关注。聚糖可以通过质谱法检测到,但最近基质辅助激光解吸/电离-质谱成像(MALDI-MSI)已经能够可视化组织上聚糖的分布。在这项研究中,我们专注于唾液酸化聚糖(唾液酸聚糖),研究了用于可视化聚糖分布的酰胺化反应,并开发了一种通过苄基酰胺化来衍生唾液酸的方法,该方法比传统的酰胺化更灵敏。此外,我们使用 MALDI-MSI 将该方法适应于可视化正常小鼠和非酒精性脂肪性肝炎(NASH)模型小鼠的福尔马林固定石蜡包埋(FFPE)肝组织中的聚糖。结果,在 NASH 小鼠肝脏中观察到糖蛋白 N-乙酰神经氨酸(NeuAc)离子分布增加,并阐明了 NASH 模型中聚糖结构的变化。