title: Alkali burn causes aldehyde dehydrogenase 3A1 (ALDH3A1) decrease in mouse cornea.
abstract: OBJECTIVE
  Aldehyde dehydrogenase 3A1 (ALDH3A1) is the most abundant soluble protein component in the mouse cornea, produced mainly by corneal epithelial cells. High levels of ALDH3A1 in cornea contribute to maintenance of a stable an d transparent corneal structure. Alkali burn is a common damage to the corneal surface, which produces an alkaline hydrolysis of matrix proteins and induces an inflammatory reaction. Our study was intended to detect changes in ALDH3A1 expression after corneal alkaline burn.
  
  
  METHODS
  To address this issue we employed RTQ-PCR to monitor the transcriptional change of ALDH3A1 after alkali burn. We used zymography to test enzyme activity changes of ALDH3A1 in the alkali burn cornea; And SDS-PAGE and mass spectrometry technology were used to verify protein content changes and to identify ALDH3A1 protein.
  
  
  RESULTS
  Using zymography, ALDH3A1 enzymic activity was observed to decrease immediately after corneal alkali burn and the levels recovered following healing. Proteins extracted from alkali burned corneas, when run on SDS-PAGE, showed the same sized band (about 54 kDa, which is the molecular weight of ALDH3A1) but in much smaller quantity, compared to normal corneas. This result was further verified by mass spectrometry fingerprinting of the in-gel lysis product. An immediate decrease of ALDH3A1 transcription after alkali burning of the cornea was also found using RTQ-PCR. This level of transcription was gradually restored during healing.
  
  
  CONCLUSIONS
  Alkali burn of the corneal surface caused a rapid decrease of ALDH3A1 in the corneal at both the RNA and protein levels, which leads to the loses of the protective component of the corneal surface and makes it vulnerable to further damage. The ALDH3A1 level in the cornea gradually recovered during the healing process. Use of an anti-oxidation reagent as a treatment ingredient for alkali burn of the corneal surface could compensate for the decrease of anti-oxidation protection potential caused by ALDH3A1 loss.
authors.affiliation: Genetic Engineering Lab, Beijing Institute of Biotechnology, Beijing, Peoples Republic of China. diceryi@hotmail.com
authors.forename: Yi
authors.initials: Y
authors.lastname: Feng
authors.forename: Yumei
authors.initials: Y
authors.lastname: Feng
authors.forename: Xudong
authors.initials: X
authors.lastname: Zhu
authors.forename: Ying
authors.initials: Y
authors.lastname: Dang
authors.forename: Qingjun
authors.initials: Q
authors.lastname: Ma
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002057:Burns, Chemical / Q000201:enzymology*; D003316:Corneal Diseases / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D005126:Eye Burns / Q000139:chemically induced*; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D010449:Peptide Mapping; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012972:Sodium Hydroxide / Q000633:toxicity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000444:Aldehyde Dehydrogenase; D005136:Eye Proteins; D012333:RNA, Messenger; D012972:Sodium Hydroxide; C105543:Aldh3a1 protein, mouse