Ascl1-mediated enhancement of GABAergic neuronal function in differentiated F11 cells under high glucose conditions.

Gamma-aminobutyric acid (GABA)ergic neurons play a key role in pain modulation within the dorsal root ganglion (DRG), making them critical targets for therapeutic studies. This study utilized F11 cells as an in vitro model to examine GABAergic function under high-glucose conditions mimicking diabetic neuropathy. Differentiated F11 cells exhibited increased sensory neuronal marker expression and functional action potentials. Overexpression of the transcription factor Achaete-scute homolog 1 (Ascl1) via lentiviral vectors enhanced GABAergic characteristics, including upregulation of GAD65, GAD67, VGAT, and GABA release. Under high-glucose conditions, Ascl1 modulated pro-inflammatory cytokines (TNF-α, NF-κB, IL-1β), anti-inflammatory cytokines (IL-4, IL-10), and pain-related channels (TRPV1, TRPA1, Nav1.8), reversing pathological changes. Temporal control of Ascl1 during differentiation reduced hypersensitivity and improved cell viability, mediated by parvalbumin in a specific GABAergic subtype. These findings highlight the therapeutic potential of Ascl1 in neuropathic pain and the scalability of F11 cells for high-throughput screening of GABAergic therapeutics.