Reactive glia express cytosolic phospholipase A2 after transient global forebrain ischemia in the rat.
OBJECTIVE
Phospholipid breakdown has been reported to be an early event in the brain after global cerebral ischemia. Our earlier observations showing the localization of cytosolic phospholipase A2 (cPLA2) to astrocytes in aged human brains and the intense glial activation observed after global forebrain ischemia prompted us to investigate the cellular localization of cPLA2 in the rat brain subjected to global ischemia.


METHODS
Immunohistochemistry was performed in sections through the dorsal hippocampus in rats subjected to 30 minutes of four- vessel occlusion. PLA2 was localized with the use of a highly selective antiserum. Double immunofluorescent localization was performed to colocalize cPLA2 with various glial cell types. cPLA2 levels were also measured by enzymatic assay and Western blot analysis.


RESULTS
A marked induction of cPLA2 was observed in activated microglia and astrocytes in the CA1 hippocampal region at 72 hours after ischemia. Only a subset of astrocytes and microglia were immunoreactive for cPLA2. Twenty-four hours after ischemia, numerous cPLA2 immunoreactive astrocytes were observed. Western blot analysis of hippocampal homogenates at 72 hours after ischemia showed induction of a 100-kD band that comigrated with purified human cPLA2, and a threefold induction in cPLA2 activity was demonstrated by enzymatic assay.


CONCLUSIONS
These results indicate that both reactive astrocytes and microglia contain elevated levels of cPLA2. Induction of cPLA2 was confined to areas of neurodegeneration and likely precedes its onset. The results suggest that reactive glia may play a role in the pathophysiology of delayed neuronal death after transient global forebrain ischemia.
Clemens
J A
JA
Eli Lilly and Company, Indianapolis, Ind., 46285, USA.
Stephenson
D T
DT
Smalstig
E B
EB
Roberts
E F
EF
Johnstone
E M
EM
Sharp
J D
JD
Little
S P
SP
Kramer
R M
RM
D000818:Animals; D001253:Astrocytes / Q000201:enzymology; D015153:Blotting, Western; D002545:Brain Ischemia / Q000201:enzymology* / Q000235:genetics; D016923:Cell Death; D003600:Cytosol / Q000201:enzymology; D004317:Doxorubicin / Q000031:analogs & derivatives; D015971:Gene Expression Regulation, Enzymologic; D006624:Hippocampus / Q000201:enzymology; D006801:Humans; D008297:Male; D017628:Microglia / Q000201:enzymology; D009410:Nerve Degeneration; D009457:Neuroglia / Q000201:enzymology*; D010741:Phospholipases A / Q000032:analysis* / Q000235:genetics; D054467:Phospholipases A2; D016548:Prosencephalon / Q000098:blood supply* / Q000201:enzymology; D051381:Rats; D017208:Rats, Wistar
C049732:7-O-(2,6-dideoxy-2-fluorotalopyranose)adriamycinone; D004317:Doxorubicin; D010741:Phospholipases A; D054467:Phospholipases A2