Calcium-activated proteolysis in rat neocortex induced by transient focal ischemia.
Ischemia-induced elevation of intracellular calcium triggers a cascade of events which is considered to play a major role in neuronal death. One candidate to participate in this process is the calcium-sensitive protease, calpain. This protease is activated by calcium, and is capable of degrading critical cytoskeletal and regulatory proteins. In order to further elucidate the role of calpain in focal ischemic damage, the present study investigated the proteolysis of spectrin, a preferred substrate for calpain, in response to transient focal ischemia. Ischemia was induced by occluding reversibly both carotid arteries and the left middle cerebral artery for three hours in Sprague-Dawley rats. Western blotting techniques were used to identify and quantify the amounts of spectrin breakdown products (BDPs) in neocortical samples from the area destined for infarction, the peri-infarct area, and the contralateral hemisphere. Substantial increases in spectrin proteolysis were observed within the first few hours of ischemia in the areas that will undergo infarction. The increase in spectrin BDPs in these areas reached a plateau around the end of the 3 h ischemic period. In the peri-infarct zone, the levels of spectrin BDPs increased in a biphasic manner. A small to moderate increase was observed by the second hour of ischemia, followed by a larger increase between the 6th and 24th hours post-ischemia. The contralateral neocortex showed a significant increase in BDPs at 2 h after the initiation of ischemia. A smaller increase in BDPs was observed thereafter.(ABSTRACT TRUNCATED AT 250 WORDS)
Hong
S C
SC
Department of Neurological Surgery, University of Virginia Health Sciences Center, Charlottesville 22908.
Lanzino
G
G
Goto
Y
Y
Kang
S K
SK
Schottler
F
F
Kassell
N F
NF
Lee
K S
KS
D000704:Analysis of Variance; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002154:Calpain / Q000378:metabolism*; D002540:Cerebral Cortex / Q000378:metabolism*; D002544:Cerebral Infarction / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000032:analysis / Q000378:metabolism*; D004789:Enzyme Activation; D002546:Ischemic Attack, Transient / Q000378:metabolism*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012016:Reference Values; D015424:Reperfusion; D013049:Spectrin / Q000302:isolation & purification / Q000378:metabolism; D013997:Time Factors
D003598:Cytoskeletal Proteins; D013049:Spectrin; D002154:Calpain; D002118:Calcium