Effect of amphetamine and phencyclidine on DNA-binding activities of serum response and dyad symmetry elements.
Acute administration of D-amphetamine sulphate (AMPH) and (1-[1-phenylcyclohexyl]piperidine hydrochloride) (phencyclidine; PCP) produces a characteristic spatio-temporal distribution of c-Fos protein in the brain. As transcriptional mechanisms underlying the induction of c-fos gene expression may be regulated in a stimulus-specific manner, we have analyzed the binding activities of serum response element (SRE), dyad symmetry element (DSE) and calcium response element (CRE), the major regulatory sites of the c-fos promoter. Electrophoretic mobility shift showed that SRE binding activity was increased for 50-60%, 2-6h after AMPH, while treatment with PCP resulted in light decrease of SRE binding activity throughout the same time period. Co-administration of AMPH and PCP induced gradual increase of SRE binding activity, reaching maximum (86%) at 6h. Binding of nuclear proteins to DSE sequence was increased 1-2h after administration of AMPH (72-87%) and remained elevated till the end of the time window observed. PCP and AMPH/PCP caused different temporal profile of DSE binding with peak (40-54%) 4-6h after administration. In contrast, DNA-binding activity of the CRE sequences remained unchanged throughout the time period of 6h under all conditions. Finally, supershift analysis clearly demonstrated presence of SRF and c-Fos protein in the transcriptional complexes bound to SRE and DSE sequences irrespective to AMPH, PCP or combined treatment. These findings also showed that the presence of c-Fos protein in SRE and DSE nucleocomplex support the hypothesis concerning autoregulation of c-fos gene expression during psychostimulant action in vivo.
Milanović
Desanka
D
Laboratory of Molecular Neurobiology, Department of Neurobiology and Immunology, Institute for Biological Research, 29 Novembra 142, 11060 Belgrade, Yugoslavia.
Milosević
Javorina
J
Kanazir
Selma
S
Rakić
Ljubisav
L
Ruzdijić
Sabera
S
D000661:Amphetamine / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D001923:Brain Chemistry / Q000187:drug effects; D000697:Central Nervous System Stimulants / Q000494:pharmacology*; D004251:DNA Transposable Elements; D024202:Electrophoretic Mobility Shift Assay; D016762:Genes, fos / Q000235:genetics*; D006213:Hallucinogens / Q000494:pharmacology*; D008297:Male; D010622:Phencyclidine / Q000494:pharmacology*; D016760:Proto-Oncogene Proteins c-fos / Q000096:biosynthesis; D051381:Rats
D000697:Central Nervous System Stimulants; D004251:DNA Transposable Elements; D006213:Hallucinogens; D016760:Proto-Oncogene Proteins c-fos; D000661:Amphetamine; D010622:Phencyclidine