engleski

Differential display PCR : study on antidepressant-induced changes of gene expression in the rat brain

Serotonergic (5-hydroxitryptaminegric, 5HT) neurotransmission is thought to be central in the etiology of depression and in the therapeutic actions of antidepressant drugs. Some of these agents - selective serotonin reuptake inhibitors (SSRI), exert their effect by blocking the reuptake of the neurotransmitter, enhancing so the 5HT transmission at the synapse. In order to get inside into the genetic basis of pharmacodynamic mechanisms of antidepressants, differential display PCR was used to search for the genes in the rat midbrain raphe region that were specifically induced or inhibited by the acute administration of SSRI fluoxetine. Total RNA was isolated from the midbrain raphe region of the fluoxetine - trated (20 mg/kg) and control rats, reversely transcribed to cDNA, and amplified, using combinations of anchored oligo-dT and arbitrary primers, in the presence of 35S-dATP. PCR products from both cDNA samples were separated on 6 percent denaturing polyacrilamide gel, visualised by autoradiography and analyzed. 18 differnetially displayed fragments were isolated from the gel, reamplified, and their differential expression was tested by the reverse Southern blot. Identification of the three genes that appeared to be inhibited and one to be induced after the acute fluoxetine administration is now in course.

Serotonin; Antidepressant; Rat

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