Mitochondria respiratory chain related genes expression in post ischemia/reperfusion rat / 中国病理生理杂志

ABSTRACT

AIM: Apply the newly invented method of suppression subtractive hybridization to scan the mitochondria genes expression of hippocampus neurons in post ischemia/reperfusion rats. METHODS: Decapitate the rats suffering 10 min of whole brain ischemia and 24 h of reperfusion in Pulsinelli’s 4 vessels occlusion ischemia/reperfusion model. Dissociated hippocampus and isolated mRNA. After reversed transcription, use SSH to subtract the common sequences, then use suppressed PCR to amplify the differentially expressed sequences. After cloning and DNA sequencing, the dot hybridization was used to finally identify the differential gene expression. RESULTS: In the 78 positive clones acquired from cloning, 7 clones represent differential gene expression after confirmed by clone PCR, dot blot and DNA sequencing. The DNA sequencing data indicate these clones are ESTs of Cytochrome c oxidase subunit and NADH-ubiquinone oxidoreductase chain 4 gene. CONCLUSION: Cytochrome C oxidase mRNA level changs during brain ischemia /reperfusion. The differential expression of NADH-ubiquinone oxidoreductase chain 4, one of those 13 subunits of subcomplex ?, suggests that subcomplex ? may have important function associated with pathophysiologic changs in reperfusion injury.