Changes in Messenger Ribonucleic Acid Species in Gerbil Brain Perturbation

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The differential display technique was applied to observe the molecular dynamism of messenger ribonucleic acid (mRNA) in this study. Using this technique, the changes in mRNAs in brain perturba-tion such as ischemia was observed to understand the molecular base of the reaction. The transient cerebral ischemia was induced by clipping both common carotid arteries for 5 minutes in Mongolian gerbils. The total RNA was extracted from the hippocampal tissue samples before ischemia, 6 hours and 2 days after ischemia. The mRNAs were reverse transcribed and subsequently amplified by polymerase chain reaction (PCR). PCR products were displayed by autoradiography as ladders on a denaturing poly-acrylamide gel. According to the autoradiography, mRNAs were divided into three patterns: 1) mRNAs obtained from the control decreased at 6 hours after 5-minute ischemia and disappeared at 2 days com-pletely; 2) decreased mRNAs at 6 hours after ischemia recovered at 2 days; and 3) new mRNAs appeared after cerebral ischemia. Located bands of interest on a gel were cut out and reamplification of com-plementary deoxyribonucleic acid was performed. The pGEM-T Vector System was used for subcloning of the amplified PCR products. The differential display technique is the powerful method for detecting genes that are unique to ischemic processes and reactions.
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