title: Detection and quantification of sulfated disaccharides from keratan sulfate and chondroitin/dermatan sulfate during chick corneal development by ESI-MS/MS.
abstract: OBJECTIVE
  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.
authors.affiliation: Division of Biology, Kansas State University, Manhattan, KS 66506, USA. ytz@ksu.edu
authors.forename: Yuntao
authors.initials: Y
authors.lastname: Zhang
authors.forename: Abigail H
authors.initials: AH
authors.lastname: Conrad
authors.forename: Elena S
authors.initials: ES
authors.lastname: Tasheva
authors.forename: Ke
authors.initials: K
authors.lastname: An
authors.forename: Lolita M
authors.initials: LM
authors.lastname: Corpuz
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kariya
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Suzuki
authors.forename: Gary W
authors.initials: GW
authors.lastname: Conrad
mesh_terms: D000818:Animals; D002642:Chick Embryo; D002645:Chickens; D002809:Chondroitin Sulfates / Q000032:analysis*; D003315:Cornea / Q000737:chemistry* / Q000196:embryology*; D003871:Dermatan Sulfate / Q000032:analysis*; D004187:Disaccharides / Q000032:analysis*; D044927:Electron-Transferring Flavoproteins; D007632:Keratan Sulfate / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D004187:Disaccharides; D044927:Electron-Transferring Flavoproteins; D003871:Dermatan Sulfate; D002809:Chondroitin Sulfates; D007632:Keratan Sulfate