Neuronal cell lines transfected with the dopamine D2 receptor gene promoter as a model for studying the effects of antidepressant drugs.
The present study investigated the effect of three antidepressant drugs (ADs), desipramine (DMI, a noradrenaline reuptake inhibitor), citalopram (CIT, a selective serotonin reuptake inhibitor) and mianserin (MIA, thought to act as an antagonist of pre-synaptic alpha2 adrenoceptor) on the transcriptional activity of the dopamine D2 receptor gene promoter. The fragment of dopamine D2 receptor gene promoter (-850 to +133) was subcloned into pGL3 vector (Promega), which has an insert coding for luciferase used as a reporter gene. Such construct (pGL3-D2R) was used to transiently transfect the neuroblastoma cell lines, Neuro 2a, SH-SY5Y and NB41A3, which endogenously express the dopamine D2 receptor protein. The obtained results indicate that transcriptional activity of dopamine D2 receptor gene promoter was dose-dependently increased by retinoic acid, forskolin, rolipram and phorbol 12 myristate 13-acetate, as well as by DMI, CIT and MIA. In the Neuro 2a cells, the most significant increase was observed after the ADs were present in the incubation medium at a doses of 0.1-1 microM for 72 h. In the SH-SY5Y cells, the significant increase in the transcriptional activity of D2 receptor gene promoter was observed already after 24-h exposure to DMI. Incubation of the Neuro 2a cells in the presence of forskolin (1 microM) or rolipram (50 microM) (but not phorbol 12-myristate 13-acetate at 0.1 microM) in combination with DMI resulted in the further increase in transcriptional activity of the studied promoter, indicating the involvement of protein kinase A pathway in these effects.
Dziedzicka-Wasylewska
Marta
M
Department of Pharmacology, Institute of Pharmacology, Polish Academy of Sciences, 12 Smetna Street, PL 31-343, Cracow, Poland. wasyl@if-pan.krakow.pl
Solich
Joanna
J
D000818:Animals; D000928:Antidepressive Agents / Q000378:metabolism / Q000494:pharmacology*; D045744:Cell Line, Tumor; D005576:Colforsin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D009447:Neuroblastoma; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D011401:Promoter Regions, Genetic; D051381:Rats; D017448:Receptors, Dopamine D2 / Q000235:genetics* / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic; D014162:Transfection; D014212:Tretinoin / Q000494:pharmacology
D000928:Antidepressive Agents; D017448:Receptors, Dopamine D2; D005576:Colforsin; D014212:Tretinoin; D013755:Tetradecanoylphorbol Acetate