TRPA1 and TRPV1 channels respond to external stimulation as pain mediators and form a complex with a transmembrane protein TMEM100 in some tissues. However, their expression and interaction in dental pulp is unclear. To investigate the functional co-expression of TRPA1 channel, TRPV1 channel and TMEM100 in human odontoblasts (HODs), immunohistochemistry, immunofluorescence staining and Western blot were used to study their co-localization and expression in both native HODs and cultured HOD-like cells. Calcium imaging was used to detect the functional interaction between TRPA1 and TRPV1 channels. Immunohistochemistry and multiple immunofluorescence staining of tooth slices showed positive expression of TRPA1 channel, TRPV1 channel and TMEM100 mainly in the cell bodies of HODs, and TRPA1 channel presented more obvious immunofluorescence in the cell processes than TRPV1 channel and TMEM100. HALO software analysis showed that TRPA1 and TRPV1 channels were positively expressed in most TMEM100⁺ HODs and these three proteins were strongly correlated in HODs (P < 0.01). The protein expression levels of TRPA1 channel, TRPV1 channel and TMEM100 in HODs showed no significant difference (P > 0.05). Double immunofluorescence staining of cultured HOD-like cells visually demonstrated that TRPA1 and TRPV1 channel were both highly co-localized with TMEM100 with similar expressive intensity. Calcium imaging showed that there was a functional interaction between TRPA1 and TRPV1 channels in HOD-like cells, and TRPA1 channel might play a greater role in this interaction. Overall, we concluded that TRPA1 channel, TRPV1 channel and TMEM100 could be functionally co-expressed in HODs.

TRPA1 和 TRPV1 通道作为疼痛介质响应外部刺激，并在某些组织中与跨膜蛋白 TMEM100 形成复合物。然而，它们在牙髓中的表达和相互作用尚不清楚。为了研究 TRPA1 通道、TRPV1 通道和 TMEM100 在人成牙本质细胞 (HOD) 中的功能共表达，使用免疫组织化学、免疫荧光染色和蛋白质印迹来研究它们在天然 HOD 和培养的 HOD 样细胞中的共定位和表达。钙成像用于检测 TRPA1 和 TRPV1 通道之间的功能相互作用。牙片免疫组化和多重免疫荧光染色显示TRPA1通道、TRPV1通道和TMEM100主要在HODs细胞体中阳性表达，并且TRPA1通道在细胞过程中呈现出比TRPV1通道和TMEM100更明显的免疫荧光。HALO 软件分析显示 TRPA1 和 TRPV1 通道在大多数 TMEM100 中呈阳性表达⁺ HODs 与这三种蛋白质在 HODs 中呈强相关性 ( P  < 0.01)。HODs中TRPA1通道、TRPV1通道和TMEM100蛋白表达水平差异无统计学意义（P  > 0.05）。培养的 HOD 样细胞的双重免疫荧光染色直观地表明 TRPA1 和 TRPV1 通道均与 TMEM100 高度共定位，具有相似的表达强度。钙成像显示HOD样细胞中TRPA1和TRPV1通道之间存在功能相互作用，TRPA1通道可能在这种相互作用中发挥更大的作用。总体而言，我们得出结论，TRPA1 通道、TRPV1 通道和 TMEM100 可以在 HOD 中功能共表达。