Pharmacological Profile of Brain-derived Neurotrophic Factor (BDNF) Splice Variant Translation Using a Novel Drug Screening Assay

• DOI: 10.1074/jbc.m114.586719
• Publication Date: 2014-07-30T04:05:02Z
• Authors: Valentina Vaghi; Alessio Polacchini; Gabriele Baj; Vera L.M. Pinheiro; Annalisa Vicario; Enrico Tongiorgi
• Keywords: splice

The neurotrophin brain-derived neurotrophic factor (BDNF) is a key regulator of neuronal development and plasticity. BDNF major pharmaceutical target in neurodevelopmental psychiatric disorders. However, pharmacological modulation this challenging because generated by multiple, alternatively spliced transcripts with different 5′- 3′UTRs. Each mRNA variant transcribed independently, but translation regulation unknown. To evaluate the translatability transcripts, we developed an vitro luciferase assay human neuroblastoma cells. In unstimulated cells, each 3′UTR determined basal level reporter gene. constructs either 5′UTR or alone showed poor BDNF, KCl, dihydroxyphenylglycine, AMPA, NMDA, dopamine, acetylcholine, norepinephrine, serotonin. Constructs consisting gene flanked one most abundant brain (exons 1, 2c, 4, 6) long responded selectively to stimulation receptor agonists, only 2c 6 were increased antidepressants desipramine mirtazapine. We propose that variants represent "a quantitative code" for regulated expression protein. Thus, discriminate efficacy drugs stimulating synthesis, it appropriate use variant-specific screening tests.