In vivo evaluation of platelet--endothelial interactions after transient retinal ischemia.
OBJECTIVE
Accumulating evidence suggests that platelets play an important role in ischemia-reperfusion injury. To fulfill that role, platelets flowing in the bloodstream would have to interact with retinal endothelial cells and to accumulate in the postischemic retina. This study was designed to investigate quantitatively platelet-endothelial interactions in postischemic retina after transient retinal ischemia.


METHODS
Transient retinal ischemia was induced in Long-Evans rats for 60 minutes by temporal ligation of the optic nerve. Isolated platelet samples labeled with carboxyfluorescein diacetate succinimidyl ester were administered intravenously to recipient rats after various reperfusion periods. Platelet-endothelial interactions in postischemic retina were evaluated in vivo with a scanning laser ophthalmoscope. Anti-P-selectin monoclonal antibody (mAb) was administered 5 minutes before the injection of labeled platelets. P-selectin gene expression in the postischemic retina was studied by semiquantitative polymerase chain reaction.


RESULTS
Under basal conditions, infused platelets showed minimal interactions with retinal endothelial cells. In contrast, postischemic retinas showed active platelet-endothelial interactions. Many platelets were observed rolling along and adhering to the major retinal veins. The number of rolling and adhering platelets reached a peak (555 +/- 65/mm per min and 25.8 +/- 3.2/mm(2)) 12 hours after reperfusion. However, the interactions between platelets and postischemic retinal endothelial cells were substantially inhibited by neutralizing P-selectin expressed on endothelial cells. In addition, P-selectin gene expression in postischemic retina corresponded with the time course of platelet-endothelial interactions during the reperfusion period.


CONCLUSIONS
This study demonstrated that platelets actively interacted with retinal endothelial cells in the postischemic retina through P-selectin expressed on the retinal endothelial cells.
Nishijima
K
K
Department of Ophthalmology and Visual Sciences, Kyoto University Graduate School of Medicine, Japan.
Kiryu
J
J
Tsujikawa
A
A
Honjo
M
M
Nonaka
A
A
Yamashiro
K
K
Tanihara
H
H
Tojo
S J
SJ
Ogura
Y
Y
Honda
Y
Y
D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D002448:Cell Adhesion; D004730:Endothelium, Vascular / Q000378:metabolism*; D005452:Fluoresceins; D005456:Fluorescent Dyes; D015870:Gene Expression; D007091:Image Processing, Computer-Assisted; D008297:Male; D009887:Ophthalmoscopy; D019007:P-Selectin / Q000235:genetics; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D020318:Rats, Long-Evans; D015427:Reperfusion Injury / Q000378:metabolism*; D012164:Retinal Diseases / Q000378:metabolism*; D012171:Retinal Vessels / Q000378:metabolism
D005452:Fluoresceins; D005456:Fluorescent Dyes; D019007:P-Selectin; D012333:RNA, Messenger; C024098:6-carboxyfluorescein