Related Experiment Videos
Central administration of dopamine D3 receptor antisense to rat: effects on locomotion, dopamine release and
A Ekman1, H Nissbrandt, M Heilig
1Institute of Physiology and Pharmacology, Department of Pharmacology, Göteborg University, Sweden.
Abstract:
A 15-mer, all-phosphorothioate-modified antisense oligodeoxynucleotide (ASO) targeted against rat dopamine D3 receptor mRNA (4 microM, 5 days) significantly reduced (28%) the amount of binding sites labelled with [3H]spiperone in monolayer cultured Chinese hamster ovary (CHO) cells transfected with the complementary desoxyribonucleic acid (cDNA) for the rat D3 receptor. In contrast, D3-ASO treatment did not reduce the amount of bound [3H]spiperone in CHO cells transfected with D2 receptor cDNA. Intracerebroventricular infusion of D3-ASO (osmotic minipump, 10 microg/microl/h, 7 days) influenced dopamine receptor density in the limbic forebrain such that the upper part of the dopamine/[3H]spiperone displacement curve--tentatively representing the D3 receptor--was altered significantly. Spontaneous locomotor activity of non-habituated rats was increased significantly in D3-ASO-treated animals; in addition, in vivo microdialysis revealed a moderate increase in dopamine release in the nucleus accumbens in these animals. In all experiments, an oligodeoxynucleotide comprising the same nucleotides as the antisense sequence, but in random order, was used as control. It is concluded that the antisense strategy is useful for investigating the functional role of dopamine D3 receptors and that the dopamine D3 receptor is involved in rat locomotor behaviour.
Insights
Antisense oligodeoxynucleotides targeting the dopamine D3 receptor reduced receptor binding and altered dopamine release in rats. This suggests the dopamine D3 receptor plays a role in regulating locomotor behavior.
Area of Science:
- Neuroscience
- Molecular Biology
- Pharmacology
Background:
- Dopamine receptors, particularly D3, are implicated in behavior.
- Antisense oligodeoxynucleotides (ASOs) offer a method to selectively target gene expression.
Purpose of the Study:
- To investigate the functional role of the dopamine D3 receptor using an antisense strategy.
- To determine the effect of dopamine D3 receptor modulation on rat behavior and dopamine release.
Main Methods:
- Treatment of cultured cells and rats with a phosphorothioate-modified antisense oligodeoxynucleotide (ASO) targeting rat dopamine D3 receptor mRNA.
- Measurement of [3H]spiperone binding to dopamine receptors in cell cultures.
- Intracerebroventricular infusion of D3-ASO in rats.
- Assessment of spontaneous locomotor activity and in vivo microdialysis of dopamine release in the nucleus accumbens.
Main Results:
- D3-ASO significantly reduced dopamine D3 receptor binding sites in transfected cells.
- Intracerebroventricular D3-ASO altered dopamine receptor density in the rat limbic forebrain.
- D3-ASO treatment increased spontaneous locomotor activity and dopamine release in the nucleus accumbens in rats.
Conclusions:
- The antisense strategy is effective for studying the function of dopamine D3 receptors.
- The dopamine D3 receptor is involved in the regulation of rat locomotor behavior.