Detection and quantification of sulfated disaccharides from keratan sulfate and chondroitin/dermatan sulfate during chick corneal development by ESI-MS/MS.
Detection and quantification of sulfated disaccharides from keratan sulfate and chondroitin/dermatan sulfate during chick corneal development by ESI-MS/MS.
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DOI：
10.1167/iovs.04-1453
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发表时间：
2005-05
影响因子：
4.4
通讯作者：
Yuntao Zhang;A. H. Conrad;E. Tasheva;K. An;L. Corpuz;Y. Kariya;Kiyoshi Suzuki;G. Conrad
中科院分区：
文献类型：
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作者：
Yuntao Zhang;A. H. Conrad;E. Tasheva;K. An;L. Corpuz;Y. Kariya;Kiyoshi Suzuki;G. Conrad
目的应用电喷雾串联质谱（ESI-MS/MS）技术鉴定和定量鸡角膜发育过程中硫酸角质素（KS）和硫酸软骨素/硫酸皮肤素（CS/DS）硫酸化二糖的变化。方法取白色来航鸡胚胎第8 ~ 20天、4周龄和70周龄的新鲜非固定角膜冰冻切片。将载玻片上的组织切片与选定的糖苷酶一起孵育。结果E8角膜中KS单硫酸化二糖（MSD）Gal-β-1，4-GlcNAc（6S）的浓度与E20相当，E10降至最低水平，E14达到第二个峰值，E18降至第二个低点，E20再次达到峰值，成人角膜仍保持较高水平。对于KS二硫酸化二糖（DSD）Gal（6S）-β-1，4-GlcNAc（6S），观察到类似的浓度曲线，因此对于总硫酸化KS二糖也是如此。在E8 ~ E18，DSD的摩尔百分比高于MSD，E20时与MSD相当，而在成人角膜中，DSD的摩尔百分比低于MSD。相比之下，CS/DS Deltadi-4S加Deltadi-6S的总浓度随着发育进展而降低，并且在成人角膜中最低。Deltadi-6S的浓度和摩尔百分比在E8时最高，然后随着Deltadi-4S的浓度和摩尔百分比从E8开始增加并在E14后超过Deltadi-6S的浓度和摩尔百分比而通过发育降低。结论新的，快速的，直接的化学分析从胚胎和成年鸡角膜切片获得的细胞外基质成分揭示了迄今未检测到的KS和CS/DS二糖在角膜发育过程中的硫酸化特性的变化。
PURPOSE To identify and quantify changes in keratan sulfate (KS) and chondroitin/dermatan sulfate (CS/DS) sulfated disaccharides in the developing chick cornea using electrospray ionization tandem mass spectrometry (ESI-MS/MS). METHODS Cryostat sections of fresh nonfixed corneas were obtained from White Leghorn embryonic day (E)8 to E20 chicks, and from 4- and 70-week-old chickens. Tissue sections on glass slides were incubated with selected glycosidase enzymes. Digest solutions were analyzed directly by ESI-MS/MS. RESULTS The concentration of KS monosulfated disaccharide (MSD) Gal-beta-1,4-GlcNAc(6S) in E8 cornea equaled that at E20, declined to its lowest level by E10, increased to a second peak by E14, decreased to a second low by E18, peaked again by E20, and remained high in adult corneas. A similar concentration profile was observed for KS disulfated disaccharide (DSD) Gal(6S)-beta-1,4-GlcNAc(6S), and thus also for total sulfated KS disaccharides. The molar percent of DSD was higher than that of MSD from E8 to E18, equivalent at E20, and less than that of MSD in adult corneas. In contrast, total concentration of CS/DS Deltadi-4S plus Deltadi-6S decreases as development progresses and is lowest in adult corneas. Concentration and molar percent of Deltadi-6S is highest at E8, then decreases through development as the concentration and molar percent of Deltadi-4S increases from E8 and exceeds that of Deltadi-6S after E14. CONCLUSIONS New, rapid, direct chemical analysis of extracellular matrix components obtained from sections from embryonic and adult chick corneas reveals heretofore undetected changes in sulfation characteristics of KS and CS/DS disaccharides during corneal development.