Changes in messenger ribonucleic acid species in gerbil brain perturbation

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 perturbation 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 polyacrylamide 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 completely; 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 complementary 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.