Columbia Technology Ventures

Assay for neurotransmitter leakage detection using fluorescent analogues

This technology is an advanced in vitro assay leveraging false fluorescent neurotransmitters (FFNs) to detect and quantify neurotransmitter leakage from synaptic vesicles, offering a dynamic and high-throughput method with implications for understanding neurological diseases and therapeutic developments.

Unmet Need: Quantitative methodology for neurotransmitter vesicular leakage

Current methods predominantly use radioactive neurotransmitters with isolated vesicles to study vesicular uptake dynamics, leaving the leakage dynamics of neurotransmitters less explored. These radiolabeled techniques offer a static measurement at a single time point, not capturing the dynamic nature of vesicular sequestration. Moreover, they introduce hazards associated with radioactive substances and lack physiological relevance as they study isolated vesicles absent from their cellular context. There is a need for a safer, real-time, and physiologically relevant approach.

The Technology: Real-time assay for neurotransmitter leakage using fluorescent analogues

The technology employs HEK293 cells stably expressing human vesicular monoamine transporter 2 (VMAT2) and human SV2C, combined with a fluorescent dopamine analogue, FFN206. This setup facilitates monitoring of vesicular retention and leakage of neurotransmitters in real time. The addition of inhibitors, like tetrabenazine (TBZ), allows for the cessation of reuptake after leakage, enabling precise quantification of leakage dynamics. By avoiding the use of radioactive substances, this method is both safer and offers the potential for multiple measurements over time for each sample, capturing the dynamic nature of neurotransmitter sequestration.

This technology has been validated in HEK293 cells and can be expanded to other cell lines, primary cultures, pluripotent stem cells, and ex vivo preparations.

Applications:

  • Diagnostic tool to study genetic and environmental factors influencing neurotransmitter leakage
  • Research platform for understanding neurotransmitter sequestration and leakage
  • Drug screening for neurological disorders like Parkinson’s disease
  • Evaluation of environmental toxins on vesicular leakage
  • Pharmacological research in vesicular dynamics

Advantages:

  • Real-time monitoring of neurotransmitter dynamics
  • Avoids hazardous radioactive substances
  • Physiologically relevant approach
  • Suitable for high-throughput screening
  • Adaptable across various cell types

Lead Inventor:

Gary Miller, Ph.D.

Patent Information:

Patent Pending

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