title: Expression of phosphorylated heat shock protein 27 during corneal epithelial wound healing.
abstract: OBJECTIVE
  The 27-kDa heat shock protein (HSP27) has been implicated in wound healing in multiple tissues. We investigated the expression and localization of phosphorylated HSP27 during epithelial wound healing in the murine cornea.
  
  
  METHODS
  Corneas of 8- to 10-week-old C57BL6 mice were wounded by epithelial debridement (n = 40). Unwounded corneas served as controls (n = 3). After 3, 7, and 14 days, phosphorylated HSP27 localization in wounded corneas was observed by confocal immunohistochemistry and double immunogold labeling transmission immunoelectron microscopy. Western blot analysis was performed to determine expression levels of phosphorylated HSP27 in scraped epithelia. Phosphorylated HSP27 localization was also separately performed with confocal immunohistochemistry 8 hours after epithelial debridement to investigate the early epithelial wound-healing process.
  
  
  RESULTS
  In unwounded corneas, phosphorylated HSP27 was localized only to the superficial epithelium. In contrast, phosphorylated HSP27 was localized in the basal and superficial epithelia 3 days after corneal epithelial wounding. After 7 and 14 days, HSP27 localization was similar to that in unwounded controls. Expression levels of phosphorylated HSP27 were greater in wounded corneal epithelia on day 3 than in unwounded controls and on day 14. After 8 hours, phosphorylated HSP27 expression was prominent in the leading edge of migrating corneal epithelium.
  
  
  CONCLUSIONS
  Constitutive expression of phosphorylated HSP27 is limited to the superficial corneal epithelium in unwounded murine corneas. Changes in HSP27 epithelial distribution and expression levels after corneal epithelial wounding suggest that phosphorylated HSP27 plays a role in early phase of corneal epithelial wound healing.
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Sandeep
authors.initials: S
authors.lastname: Jain
authors.forename: Jose
authors.initials: J
authors.lastname: De la Cruz
authors.forename: Eunkyo
authors.initials: E
authors.lastname: Kang
authors.forename: Takashi
authors.initials: T
authors.lastname: Kojima
authors.forename: Jin-Hong
authors.initials: JH
authors.lastname: Chang
authors.forename: Jae Yong
authors.initials: JY
authors.lastname: Kim
mesh_terms: D000818:Animals; D015153:Blotting, Western; D065306:Corneal Injuries; D003646:Debridement; D019573:Epithelium, Corneal / Q000378:metabolism* / Q000648:ultrastructure; D055551:HSP27 Heat-Shock Proteins / Q000378:metabolism*; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D016253:Microscopy, Immunoelectron; D010766:Phosphorylation; D014945:Wound Healing
chemical_list: D055551:HSP27 Heat-Shock Proteins; C486699:Hspb2 protein, mouse