Qualitative and Quantitative Changes in Proteins in Rat Cerebral Cortex after Transient Focal Ischemia
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概要
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To investigate the progression of neuronal damage after cerebral ischemia, the protein-alterations in rat cerebral cortex was qualitatively and quantitatively analyzed using a transient focal cerebral ischemia model. The cerebral cortex was fractionated into nuclear, mitochondrial, microsomal, and cytosolic fractions following transient ischemia. Proteins contained in each fraction were analyzed using sodium dodecyl sulfate polyacrylamide slab gel electrophoresis (SDS-PAGE) . Two proteins (molecular mass: 67 kDa and 80 kDa) were markedly increased in the cytosolic fraction 16 hr and 8 days following transient ischemia. Levels returned to baseline 2 weeks after lesioning. No significant differences were observed in the other fractions. The 67 kDa protein was purified by ion-exchange chromatography followed by gel permeation chromatography and partial amino acid sequences were determined. As a result, a 67 kDa protein that was increased following transient ischemia was identified as serum albumin. Moreover, in the cytosolic fraction of cerebral cortex following transient ischemia and subjection to immunoblot analysis using polyclonal antibody against rat whole serum. The results of this analysis revealed nine proteins, including the 67 and 80 kDa proteins, derived from serum. Six of these proteins showed a similar pattern of transient increase 16 hr and 8 days following transient ischemia. The 80 kDa protein was identified as transferrin in terms of its molecular mass and cross-reactivity against an anti-human transf errin polyclonal antibody.
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