P. Matz et al., SUBARACHNOID INJECTIONS OF LYSED BLOOD INDUCE THE HSP70 STRESS GENE AND PRODUCE DNA FRAGMENTATION IN FOCAL AREAS OF THE RAT-BRAIN, Stroke, 27(3), 1996, pp. 504-512
Background and Purpose Most experimental studies of subarachnoid hemor
rhage have demonstrated little histological evidence of injury. In the
present study we examined both the expression of the hsp70 heat-shock
gene, a molecular marker of reversible neuronal injury, and DNA fragm
entation, a marker of irreversible cell injury and death. Methods Lyse
d blood, whole blood, oxyhemoglobin, bovine serum albumin, and saline
were injected into the cisterna magna of adult rats. The induction of
hsp70 mRNA and HSP70 heat-shock protein was assessed with the use of i
n situ hybridization and immunocytochemistry, respectively. Fragmentat
ion of genomic DNA was studied by DNA nick end-labeling with the use o
f terminal deoxynucleotidyl transferase and biotinylated dATP. Results
Expression of the hsp70 gene was not induced in the brains of rats in
jected with whole blood, oxyhemoglobin, bovine serum albumin, or salin
e. Lysed blood injections, however, induced hsp70 mRNA at 6 and 24 hou
rs in the cerebellar hemispheres and in focal regions of the basal for
ebrain. HSP70 protein was induced by 24 hours and persisted for at lea
st 4 days in the same regions. HSP70 protein was localized to patches
of glial cells and occasional neurons in the forebrain. In the cerebel
lum HSP70 was localized to Bergmann glial cells, granule cells, molecu
lar layer stellate cells, and occasional Purkinje cells. DNA nick end-
labeling showed patches of labeled cells in the basal forebrain that o
ccurred in the same regions that hsp70 mRNA was induced. Conclusions T
he results demonstrate focal stress gene induction and DNA fragmentati
on after subarachnoid hemorrhage. it is hypothesized that the focal ar
eas of hsp70 induction may reflect ischemic injury due to vasospasm pr
oduced by lysed blood and/or injury mediated by direct toxic effects o
f the lysed blood. The hsp70 induction and DNA nick end-labeling in th
e same regions suggests that lysed blood produces a spectrum of injury
from HSP70 protein-labeled, reversibly injured cells to dead cells wi
th fragmented DNA. Induction of the hsp70 stress gene and DNA nick end
-labeling may be useful for evaluating the causes of injury, the spect
rum of injury, and potential pharmacological therapies in experimental
models of subarachnoid hemorrhage.