4187841 - Development of voltage-gated sodium channel-DART probes as a cell-specific modulator | Poster Board #827

Voltage-gated sodium (Na_v) channels are critical in initiating and propagating electrical signals in the central nervous system. Silencing neurons by manipulating Na_v channels is one of the most effective methods for identifying specific neuronal effects. Existing silencing neuronal tools exhibit limitations in a lack of cell specificity or off-target effects due to the widespread expression of Na_v channels. To address these challenges, we utilize DART (Drug Acutely Restricted by Tethering) technology developed by Tadross and colleagues, which enables spatiotemporal and cell-type-specific modulation of pharmacological targets by employing a bacterial enzyme called HaloTag. We design and synthesize novel Na_v-DART probes based on established Na_v channel blockers connected to Haltag ligands via a flexible linker to achieve a cell-specific modulation of Na_v channels. Then, we investigate the cell-specific effects of these probes on Na_v channels through whole-cell patch recordings of Na_v channel current and neurons firing in cultured hippocampal neurons and optimize their structures based on observed efficacy. Our results demonstrate that several Na_v-DART probes effectively reduce Na_v channel firing across a broad voltage range, while control probes exhibit no significant effect on Na_v channel current or firing. These newly developed Na_v-DART probes, which enable the modulation of Na_v channels with cell-type specificity, offer promising avenues for future research on animal behavior and neuronal diseases.