Involvement of calpain isoforms in ischemia-reperfusion injury in rat retina.
OBJECTIVE
Much evidence has accumulated suggesting that activation of calpain causes neuronal cell death in ischemic brain. However, little is known about the involvement of calpain in retinal cell death in ischemic injury. Thus, the purpose of present study was to investigate the involvement of calpain isoforms (m- and mu-calpain) in ischemia-reperfusion injury in retina from rat.


METHODS
Retinal ischemia was produced by occlusion of the central retinal artery for one hour, and this was followed by reperfusion for seven days. Calpain mRNAs, calpain activities, total calcium content and proteolysis of alpha-spectrin were determined in retina. Effect of a calpain inhibitor SJA6017 was histologically tested in retinal injury after ischemia-reperfusion.


RESULTS
Following retinal ischemia, most of cells in the ganglion cell layer were sloughed off by day 1 after reperfusion, followed by loss of cells in the inner plexiform layer on day 3 and loss of cells in the inner nuclear layer by day 5. These morphologic changes were accompanied by several presumptive biochemical indicators of calpain activation: increased calcium, proteolysis of alpha-spectrin (a sensitive substrate for calpains), decreased caseinolytic activity for both calpains (suggesting calpain activation followed by autolytic degradation), increased mRNA levels for mu-calpain and calpastatin - the endogenous inhibitor of calpains - and decreased mRNA levels for mu-calpain. Moreover, the calpain inhibitor SJA6017 protected the reduction of cell density in the ganglion cell layer after ischemia-reperfusion.


CONCLUSIONS
These results suggest that calpain isoforms may play an important role in neuronal cell death induced by retinal ischemia-reperfusion injury in rat.
Sakamoto
Y R
YR
Research Laboratories, Senju Pharmaceutical Corporation Limited, Kobe, Japan.
Nakajima
T R
TR
Fukiage
C R
CR
Sakai
O R
OR
Yoshida
Y R
YR
Azuma
M R
MR
Shearer
T R
TR
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002154:Calpain / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002452:Cell Count; D016923:Cell Death; D004151:Dipeptides / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007511:Ischemia / Q000473:pathology / Q000503:physiopathology*; D008297:Male; D010447:Peptide Hydrolases / Q000378:metabolism; D020033:Protein Isoforms / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000473:pathology / Q000503:physiopathology*; D012165:Retinal Ganglion Cells / Q000187:drug effects / Q000473:pathology / Q000502:physiology; D012171:Retinal Vessels; D013049:Spectrin / Q000378:metabolism
D004151:Dipeptides; D004791:Enzyme Inhibitors; C109145:N-(4-fluorophenylsulfonyl)-L-valyl-L-leucinal; D020033:Protein Isoforms; D012333:RNA, Messenger; D013049:Spectrin; D010447:Peptide Hydrolases; D002154:Calpain; D002118:Calcium