Expression of the focal adhesion protein PINCH in normal and alkali-injured corneas and the role of PMNs.
OBJECTIVE
To evaluate the role of particularly interesting new cysteine-histidine-rich protein (PINCH) in corneal wound healing and early neovascularization and to assess the influence of granulocytes.


METHODS
A standardized corneal alkali wound was inflicted under general anaesthesia to the right eye of 14 New Zealand White rabbits. Seven of the rabbits received i.v. 5 mg/kg fucoidin every 2 hours to prevent granulocytes from entering the wound area. After 36 hours, the rabbits were killed, the corneas excised, fixed in 4% formaldehyde and embedded in paraffin. The sections were double-stained with antibodies against PINCH and with haematoxylin.


RESULTS
In the normal cornea and limbus, PINCH was weakly expressed in the corneal epithelium and in a wedge of the conjunctival stroma. In the wounded corneas, PINCH expression was seen in the frontline of repopulating endothelial and epithelial cells, and in active keratocytes. The vascular endothelium and the granulocytes expressed PINCH, as did the conjunctival epithelium. In the fucoidin-treated rabbits, PINCH expression was markedly reduced. The vascular endothelial cells and the few granulocytes did not express PINCH in these rabbits.


CONCLUSIONS
PINCH is only slightly expressed in the normal cornea. A corneal wound induces PINCH expression in the repopulating cells, in the vascular endothelial cells of the limbus, in the limbal epithelium and in the granulocytes. Exclusion of granulocytes reduces expression of PINCH and there is no expression at all in the vascular endothelium.
Bourghardt Peebo
Beatrice
B
Department of Ophthalmology, University Hospital, Linköping, Sweden. beatrice.peebo@lio.se
Gan
Lisha
L
Sun
Xiao Feng
XF
Knutsen
Annica
A
Rearden
Ann
A
Fagerholm
Per
P
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D001381:Azepines; D002057:Burns, Chemical / Q000378:metabolism*; D003315:Cornea / Q000378:metabolism*; D065306:Corneal Injuries; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004195:Disease Models, Animal; D004730:Endothelium, Vascular / Q000378:metabolism; D005126:Eye Burns / Q000139:chemically induced*; D022001:Focal Adhesions / Q000378:metabolism*; D007124:Immunoenzyme Techniques; D008012:Lidocaine / Q000031:analogs & derivatives; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D011134:Polysaccharides / Q000494:pharmacology; D011817:Rabbits; D012972:Sodium Hydroxide / Q000633:toxicity; D014945:Wound Healing; D016335:Zinc Fingers
D000925:Anticoagulants; D001381:Azepines; D004268:DNA-Binding Proteins; D011134:Polysaccharides; D012972:Sodium Hydroxide; C040465:pincainide; C007789:fucoidan; D008012:Lidocaine