Differential expression of cyclooxygenase-1 and cyclooxygenase-2 in the cornea during wound healing.
Cyclooxygenases (COXs) are the key enzymes in the production of prostaglandins (PGs) and exist in two isoforms. Isoform 1 (COX-1) is constitutively expressed in most tissues, whereas cyclooxygenase-2 (COX-2) is rapidly induced by a variety of different stimuli. In this study, we have quantitatively analyzed mRNA expression of COX-1 and COX-2 and protein distribution during corneal reparative processes after wound. Total RNA was isolated from cornea samples of New Zealand rabbits that had been subjected to corneal wound by mechanical brush scraping. Quantification of RT-PCR results was made by using a DNA mimic approach. The localization and expression of the enzymes was studied by immunocytochemistry and Western blotting. In normal corneas COX-1 is expressed throughout the cornea in the whole tissue, while COX-2 is strongly expressed in stromal keratocytes. Following injury, COX-2 levels drastically increase and, at least in the epithelium, COX-2 becomes the predominant isoform of cyclooxygenases at an early stage of healing. Moreover, in the epithelium COX-2 is expressed predominantly by those cells close to the wound. These cells become migratory and move toward the injured area. In contrast, COX-1 levels remain unaffected in all corneal tissues. The system returns to the pre-injury state in about 24h. Thus, the expression of COX-2 in the corneal epithelium during wound repair is tightly regulated both temporally and spatially.
Amico
Carla
C
Department of R&D, SIFI SpA, Via Ercole Patti 36, 95020 Lavinaio, Italy. carla@sifi.it
Yakimov
Michail
M
Catania
Maria Vincenza
MV
Giuffrida
Raffaella
R
Pistone
Matteo
M
Enea
Vincenzo
V
D000818:Animals; D015153:Blotting, Western; D003315:Cornea / Q000502:physiology*; D065306:Corneal Injuries; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008861:Microsomes / Q000201:enzymology; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D014945:Wound Healing / Q000502:physiology*
D007527:Isoenzymes; D012333:RNA, Messenger; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases