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.
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 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.
Patent Pending
IR CU22294
Licensing Contact: Greg Maskel