Use of suppression subtractive hybridization strategy for discovery of increased tissue inhibitor of matrix metalloproteinase-1 gene expression in brain ischemic tolerance.
Brief occlusion of the middle cerebral artery (i.e., ischemic preconditioning; PC) induces significant brain protection to subsequent severe ischemic events. In an effort to discover genes responsible for ischemic tolerance, we have applied a new technique, suppression subtractive hybridization (SSH), to identify genes that are upregulated by PC. Using this SSH approach, a cDNA that encodes tissue inhibitor of matrix metalloproteinase- (TIMP-1) was identified. Time course studies using Northern analysis revealed that TIMP-1 mRNA was significantly elevated at 24 hours (3.3-fold over controls, P < 0.05, n = 5) and 2 days (4.3-fold increase, P < 0.01) after PC, corresponding to the onset of significant ischemic tolerance. Our data not only demonstrate the utility of this new polymerase chain reaction-based SSH strategy for discovery of genes differentially expressed in PC, but also suggest a potential role of TIMP-1 in PC-induced ischemic tolerance.
Wang
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Department of Cardiovascular Pharmacology, SmithKline Beecham Pharmaceuticals, King of Prussia, Pennsylvania 19406, USA.
Yaish-Ohad
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Li
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Barone
F C
FC
Feuerstein
G Z
GZ
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002540:Cerebral Cortex / Q000378:metabolism; D018076:DNA, Complementary; D007839:Functional Laterality; D005786:Gene Expression Regulation; D006973:Hypertension; D002546:Ischemic Attack, Transient / Q000378:metabolism*; D019194:Ischemic Preconditioning; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction; D051381:Rats; D011918:Rats, Inbred SHR; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D019715:Tissue Inhibitor of Metalloproteinase-1