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• W4322008063 abstract "Deficiencies in brain-derived neurotrophic factor (BDNF) have been linked to several brain disorders, making compounds that can boost neuronal BDNF synthesis attractive as potential therapeutics. However, a sensitive and quantitative BDNF assay for high-throughput screening (HTS) is still missing. Here we report the generation of a new mouse Bdnf allele, BdnfNLuc, in which the sequence encoding nano luciferase (NLuc) is inserted into the Bdnf locus immediately before the stop codon so that the allele will produce a BDNF-NLuc fusion protein. BDNF-NLuc protein appears to function like BDNF as BdnfNLuc/NLuc homozygous mice grew and behaved almost normally. We were able to establish and optimize cultures of cortical and hippocampal BdnfNLuc/+ neurons isolated from mouse embryos in 384-well plates. We used the cultures as a phenotypic assay to detect the ability of 10 mM KCl to stimulate BDNF synthesis and achieved a reproducible Z′ factor > 0.50 for the assay, a measure considered suitable for HTS. We successfully scaled up the assay to screen the 1280-compound LOPAC library (Library of Pharmacologically Active Compounds). The screen identified several BDNF-boosting compounds, one of which is Bay K8644, a L-type voltage-gated calcium channel (L-VGCC) agonist, which was previously shown to stimulate BDNF synthesis. These results indicate that our phenotypic neuronal assay is ready for HTS to identify novel BDNF-boosting compounds." @default.
• W4322008063 created "2023-02-26" @default.
• W4322008063 creator A5024636337 @default.
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• W4322008063 date "2023-04-01" @default.
• W4322008063 modified "2023-10-16" @default.
• W4322008063 title "High throughput assay for compounds that boost BDNF expression in neurons" @default.
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• W4322008063 doi "https://doi.org/10.1016/j.slasd.2023.02.005" @default.
• W4322008063 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/36842668" @default.
• W4322008063 hasPublicationYear "2023" @default.
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