Development of a novel TRPV1 dependent in vitro model of nociceptive pain

This study aims to develop of a novel in vitro model for pain research, focusing on the TRPV1 receptor, which plays a crucial role in pain mechanisms. Given the limitations of current therapies in treating pain sensation without targeting underlying causes, this research aims to advance understanding of TRPV1’s role in neuronal hyperexcitability and its potential as a therapeutic target. The ND7/23 cell line was transfected with human TRPV1 to create a stable neuronal model that closely mimics dorsal root ganglion neurons, known for their role in pain transmission. This model allows for a detailed investigation of TRPV1’s functionality, particularly its interactions with voltage-gated ion channels, which are key in modulating neuronal excitability and pain signalling. Capsaicin, a natural compound known for activating TRPV1, was used to evaluate the model’s effectiveness. Calcium imaging confirmed capsaicin responsiveness to TRPV1 in ND7/23-TRPV1 cells. Automated patch clamp experiments demonstrated full functionality of TRPV1 in response to heat and pH, as well as the specificity TRPV1-capsaicin binding, with the use of capsazepine. In dose response studies capsaicin demonstrated a higher potency in ND7/23-TRPV1 cells compared to CHO-TRPV1 cells, confirming its neuronal characteristics. In further studies, capsaicin highlighted its potential in modulating ionic currents and action potentials. The findings suggest that capsaicin, through TRPV1 activation, can modulate neuronal activity, offering promise as a therapeutic agent for chronic pain management. This study establishes a robust in vitro model for pain research, offering new insights into the complex mechanisms of pain. It underscores the critical role of TRPV1 in modulating neuronal excitability, laying the foundation for the development of targeted analgesic therapies.

Publisher

University of Galway

Rights

Attribution-NonCommercial-NoDerivatives 4.0 International

cbn

Collections

University of Galway Theses (Research Masters Theses)