PMID- 25074925 OWN - NLM STAT- MEDLINE DCOM- 20141217 LR - 20220408 IS - 1083-351X (Electronic) IS - 0021-9258 (Print) IS - 0021-9258 (Linking) VI - 289 IP - 40 DP - 2014 Oct 3 TI - Pharmacological profile of brain-derived neurotrophic factor (BDNF) splice variant translation using a novel drug screening assay: a "quantitative code". PG - 27702-13 LID - 10.1074/jbc.M114.586719 [doi] AB - The neurotrophin brain-derived neurotrophic factor (BDNF) is a key regulator of neuronal development and plasticity. BDNF is a major pharmaceutical target in neurodevelopmental and psychiatric disorders. However, pharmacological modulation of this neurotrophin is challenging because BDNF is generated by multiple, alternatively spliced transcripts with different 5'- and 3'UTRs. Each BDNF mRNA variant is transcribed independently, but translation regulation is unknown. To evaluate the translatability of BDNF transcripts, we developed an in vitro luciferase assay in human neuroblastoma cells. In unstimulated cells, each BDNF 5'- and 3'UTR determined a different basal translation level of the luciferase reporter gene. However, constructs with either a 5'UTR or a 3'UTR alone showed poor translation modulation by BDNF, KCl, dihydroxyphenylglycine, AMPA, NMDA, dopamine, acetylcholine, norepinephrine, or serotonin. Constructs consisting of the luciferase reporter gene flanked by the 5'UTR of one of the most abundant BDNF transcripts in the brain (exons 1, 2c, 4, and 6) and the long 3'UTR responded selectively to stimulation with the different receptor agonists, and only transcripts 2c and 6 were increased by the antidepressants desipramine and mirtazapine. We propose that BDNF mRNA variants represent "a quantitative code" for regulated expression of the protein. Thus, to discriminate the efficacy of drugs in stimulating BDNF synthesis, it is appropriate to use variant-specific in vitro screening tests. CI - (c) 2014 by The American Society for Biochemistry and Molecular Biology, Inc. FAU - Vaghi, Valentina AU - Vaghi V AD - From the Department of Life Sciences, University of Trieste, 34127 Trieste, Italy. FAU - Polacchini, Alessio AU - Polacchini A AD - From the Department of Life Sciences, University of Trieste, 34127 Trieste, Italy. FAU - Baj, Gabriele AU - Baj G AD - From the Department of Life Sciences, University of Trieste, 34127 Trieste, Italy. FAU - Pinheiro, Vera L M AU - Pinheiro VL AD - From the Department of Life Sciences, University of Trieste, 34127 Trieste, Italy. FAU - Vicario, Annalisa AU - Vicario A AD - From the Department of Life Sciences, University of Trieste, 34127 Trieste, Italy. FAU - Tongiorgi, Enrico AU - Tongiorgi E AD - From the Department of Life Sciences, University of Trieste, 34127 Trieste, Italy tongi@units.it. LA - eng GR - GGP08258/TI_/Telethon/Italy PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20140729 PL - United States TA - J Biol Chem JT - The Journal of biological chemistry JID - 2985121R RN - 0 (3' Untranslated Regions) RN - 0 (5' Untranslated Regions) RN - 0 (Antidepressive Agents) RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (RNA, Messenger) SB - IM MH - 3' Untranslated Regions MH - 5' Untranslated Regions MH - *Alternative Splicing MH - Antidepressive Agents/*pharmacology MH - Brain-Derived Neurotrophic Factor/*antagonists & inhibitors/*genetics/metabolism MH - *Drug Evaluation, Preclinical MH - Genes, Reporter MH - Humans MH - *Peptide Chain Elongation, Translational MH - RNA, Messenger/genetics/metabolism PMC - PMC4183807 OTO - NOTNLM OT - Brain-derived Neurotrophic Factor (BDNF) OT - Drug Screening OT - In Vitro Assay OT - Neurotransmitter OT - Signaling OT - Splice Variants OT - Translation EDAT- 2014/07/31 06:00 MHDA- 2014/12/18 06:00 PMCR- 2015/10/03 CRDT- 2014/07/31 06:00 PHST- 2014/07/31 06:00 [entrez] PHST- 2014/07/31 06:00 [pubmed] PHST- 2014/12/18 06:00 [medline] PHST- 2015/10/03 00:00 [pmc-release] AID - S0021-9258(20)48406-6 [pii] AID - M114.586719 [pii] AID - 10.1074/jbc.M114.586719 [doi] PST - ppublish SO - J Biol Chem. 2014 Oct 3;289(40):27702-13. doi: 10.1074/jbc.M114.586719. Epub 2014 Jul 29.