Gangliosides are sialic acid-containing glycosphingolipids characterized by their diverse carbohydrate structures and are divided into four groups such as o-, a-, b- and c-series. We reported that the gene expression of several glycosyltransferases in dorsal root ganglia (DRG) and the spinal cord are changed during inflammatory pain. In this study, we focused on the upregulation of st8sia1 which produces the starting molecule in b-series ganglioside biosynthesis in DRG, since intraplantar injection of GT1b, a b-series ganglioside, caused mechanical allodynia. GT1b-induced allodynia was blocked by TRPV1 antagonists and GT1b increased capsaicin-induced nociceptive behavior. Furthermore, GT1b affected mTRPV1-EGFP internalization in F11 cells (a DRG neuron-derived cell line). Glycosphingolipids are enriched in lipid rafts which modulate membrane protein interaction, activity and localization. Thus, it is possible that GT1b affect mTRPV1 by modulation of lipid raft function. Intraplantar injection of Methyl-β-cyclodextrin that disrupts lipid rafts attenuated GT1b-induced mechanical allodynia. These results suggested that GT1b changed the carbohydrate chain composition in lipid rafts and then induced mechanical allodynia via the TRPV1 regulation.