Perinatal hypoxia and reoxygenation (HYP/REOX) induced in cerebral tissue the urokinase type plasminogen activator (UFA) and matrix metalloproteinases 2 and 9

Alan (Roger) Hohimer

Research output: Contribution to journalArticle

Abstract

Perinatal HYP/REOX is associated with cerebral hemorrhagic events but the mechanisms which actually elicit the bleeds remain unclear. To test the hypothesis that HYP/REOX generates proteolytic agents which weaken the extracellular matrix, 5 two day old mice (from 2 different litters) were exposed to 5% O2 for 8 hours after which the pups were returned to room air for 4 hours and then were euthanized. From these pups and from 4 normoxic control mice (from 2 litters) brain tissue (cortex, striatum and thalamus) was harvested. Soluble protein extracts were made and equal amounts of protein from each group were subjected to SDS PAGE Western blot analysis utilizing chemiluminescent detection. Immunoblots evaluated with an anti-UPa antibody (American Diagnostics) showed that HYP/REOX induced a 27 Kd species of UPA. Gelatin zymography (SDS PAGE gels with 0.2% gelatin were washed briefly with Triton 100 and incubated at 37° C for 12 hours) revealed gelatinolytic activity at both 68 and 88 kD in the HYP/REOX group strongly suggesting induction of the matrix metalloproteinases 2 and 9. These proteolytic agents may be involved in the genesis of perinatal cerebral bleeds and might be appropriate targets for improved pharmacologie therapy.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number3
StatePublished - 1996

Fingerprint

t-plasminogen activator
u-plasminogen activator
gelatinase A
gelatinase B
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9
Urokinase-Type Plasminogen Activator
Tissue Plasminogen Activator
Gelatin
hypoxia
Tissue
gelatin
litters (young animals)
pups
Polyacrylamide Gel Electrophoresis
Anti-Idiotypic Antibodies
Brain
Proteins
Gels
thalamus

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

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abstract = "Perinatal HYP/REOX is associated with cerebral hemorrhagic events but the mechanisms which actually elicit the bleeds remain unclear. To test the hypothesis that HYP/REOX generates proteolytic agents which weaken the extracellular matrix, 5 two day old mice (from 2 different litters) were exposed to 5{\%} O2 for 8 hours after which the pups were returned to room air for 4 hours and then were euthanized. From these pups and from 4 normoxic control mice (from 2 litters) brain tissue (cortex, striatum and thalamus) was harvested. Soluble protein extracts were made and equal amounts of protein from each group were subjected to SDS PAGE Western blot analysis utilizing chemiluminescent detection. Immunoblots evaluated with an anti-UPa antibody (American Diagnostics) showed that HYP/REOX induced a 27 Kd species of UPA. Gelatin zymography (SDS PAGE gels with 0.2{\%} gelatin were washed briefly with Triton 100 and incubated at 37° C for 12 hours) revealed gelatinolytic activity at both 68 and 88 kD in the HYP/REOX group strongly suggesting induction of the matrix metalloproteinases 2 and 9. These proteolytic agents may be involved in the genesis of perinatal cerebral bleeds and might be appropriate targets for improved pharmacologie therapy.",
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AB - Perinatal HYP/REOX is associated with cerebral hemorrhagic events but the mechanisms which actually elicit the bleeds remain unclear. To test the hypothesis that HYP/REOX generates proteolytic agents which weaken the extracellular matrix, 5 two day old mice (from 2 different litters) were exposed to 5% O2 for 8 hours after which the pups were returned to room air for 4 hours and then were euthanized. From these pups and from 4 normoxic control mice (from 2 litters) brain tissue (cortex, striatum and thalamus) was harvested. Soluble protein extracts were made and equal amounts of protein from each group were subjected to SDS PAGE Western blot analysis utilizing chemiluminescent detection. Immunoblots evaluated with an anti-UPa antibody (American Diagnostics) showed that HYP/REOX induced a 27 Kd species of UPA. Gelatin zymography (SDS PAGE gels with 0.2% gelatin were washed briefly with Triton 100 and incubated at 37° C for 12 hours) revealed gelatinolytic activity at both 68 and 88 kD in the HYP/REOX group strongly suggesting induction of the matrix metalloproteinases 2 and 9. These proteolytic agents may be involved in the genesis of perinatal cerebral bleeds and might be appropriate targets for improved pharmacologie therapy.

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