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Related Experiment Video

Updated: Aug 8, 2025

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High throughput assay for compounds that boost BDNF expression in neurons.

Guey-Ying Liao1, Haifei Xu1, Justin Shumate2

  • 1Department of Neuroscience, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology, University of Florida, 130 Scripps Way, Jupiter, FL 33458, USA.

SLAS Discovery : Advancing Life Sciences R & D
|February 26, 2023
PubMed
Summary

Researchers developed a novel assay to measure brain-derived neurotrophic factor (BDNF) production in neurons. This high-throughput screening method successfully identified compounds that can boost BDNF synthesis, offering potential new treatments for brain disorders.

Keywords:
Brain-derived neurotrophic factorGene expressionHTS assayKnockinLOPAC libraryNano luciferase

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Drug Discovery

Background:

  • Deficiencies in brain-derived neurotrophic factor (BDNF) are implicated in various neurological disorders.
  • Developing therapeutics that enhance BDNF synthesis is a key goal in neuroscience.
  • A sensitive, quantitative assay for high-throughput screening (HTS) of BDNF-boosting compounds is currently lacking.

Purpose of the Study:

  • To generate a novel mouse model for studying BDNF synthesis.
  • To develop and validate a high-throughput screening assay for identifying compounds that increase neuronal BDNF production.
  • To screen a compound library for novel BDNF-boosting agents.

Main Methods:

  • Generation of a new mouse allele, BdnfNLuc, producing a BDNF-nano luciferase fusion protein.
  • Establishment and optimization of primary cortical and hippocampal neuron cultures for phenotypic assays.
  • Validation of the assay for HTS with a Z' factor > 0.50 and screening of the LOPAC library.

Main Results:

  • The BdnfNLuc mice exhibited normal growth and behavior, indicating functional BDNF-NLuc fusion protein.
  • The developed neuronal assay demonstrated reproducibility and suitability for HTS.
  • Screening of the LOPAC library identified several compounds that stimulate BDNF synthesis, including the L-type voltage-gated calcium channel agonist Bay K8644.

Conclusions:

  • The novel BdnfNLuc mouse model and the associated phenotypic neuronal assay are effective for HTS.
  • This assay platform is ready for screening large compound libraries to discover novel therapeutics for neurological conditions by boosting BDNF.
  • The findings validate the use of NLuc reporter systems for quantitative assessment of neurotrophic factor synthesis in drug discovery.