IOVS Journal of Bacteriology
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(Investigative Ophthalmology and Visual Science. 2003;44:3332-3338.)
© 2003 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.02-1280
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Up-Regulation of Urokinase-Type Plasminogen Activator in Corneal Epithelial Cells Induced by Wounding

Masanao Watanabe,1 Wataru Yano,1 Shoichi Kondo,1 Yukio Hattori,1 Naoyuki Yamada,2 Ryoji Yanai,2 and Teruo Nishida2

1From the Tokyo New Drug Research Laboratories, Kowa Company Ltd., Tokyo, Japan; and the 2Department of Biomolecular Recognition and Ophthalmology, Yamaguchi University School of Medicine, Yamaguchi, Japan.

PURPOSE. To investigate the possible role of urokinase-type plasminogen activator (uPA) in corneal epithelial wound healing by examining its expression both in the rabbit corneal epithelium in situ and in rabbit corneal epithelial (RCE) cells in vitro.

METHODS. The rabbit cornea was subjected to mechanical wounding, and frozen sections of the tissue were subsequently prepared and subjected both to immunostaining with antibodies to uPA and to in situ zymography for the detection of PA activity. RCE cell monolayers were also subjected to scrape wounding, after which they were immunostained for uPA. The amounts of uPA protein in the culture medium and of uPA mRNA in cell lysates were also determined by enzyme-linked immunosorbent assay and by reverse transcription and real-time quantitative polymerase chain reaction analysis, respectively.

RESULTS. Immunostaining and in situ zymography of the wounded cornea revealed that uPA was restricted to the leading edge of the migrating corneal epithelium. In contrast, tissue-type PA was expressed throughout the corneal epithelium. Scraping of RCE cell monolayers induced the expression of uPA in the migrating cells at the wound edge. The amount of uPA in the culture medium of RCE cells increased with the number of scrape wounds applied. Wounding also induced a time-dependent increase in the abundance of uPA mRNA in the cell monolayers. The migration of RCE cells was inhibited by antibodies to uPA.

CONCLUSIONS. Mechanical wounding induces up-regulation of uPA at both the protein and mRNA levels in corneal epithelial cells. uPA may thus contribute to epithelial cell migration during corneal epithelial wound healing.




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