神经酰胺
再摄取
细胞生物学
多巴胺
多巴胺转运体
鞘磷脂磷酸二酯酶
化学
激酶
多巴胺质膜转运蛋白
鞘磷脂
蛋白激酶A
多巴胺能
生物
生物化学
内分泌学
血清素
细胞凋亡
受体
膜
作者
Jong Hoon Won,Seok Kyun Kim,In Chul Shin,Hae Chan Ha,Ji Min Jang,Moon Jung Back,Dae Kyong Kim
标识
DOI:10.1016/j.cellsig.2018.01.006
摘要
Dopamine (DA) reuptake is the primary mechanism to terminate dopaminergic transmission in the synaptic cleft. The dopamine transporter (DAT) has an important role in the regulation of DA reuptake. This study provides anatomical and physiological evidence that DAT recycling is regulated by ceramide kinase via the sphingomyelin pathway. First, the results show that DAT and neutral sphingomyelinase 2 (nSMase2) were successfully co-precipitated from striatal samples and were colocalized in the mouse striatum or PC12 cells. We also identified a protein-protein interaction between nSMase2 and DAT through in situ proximity ligation assay experiments in the mouse striatum. Second, dopamine (DA) stimulated the formation of ceramide and increased nSMase activity in PC12 cells, while treatment with a cell-permeable ceramide-1-phosphate (C1P) increased DA uptake. Third, we used inhibitors and siRNA to inhibit nSMase2 and ceramide kinase and observed the effects on DAT recycling in PC12 cells. Treatment with ceramide kinase inhibitor K1, or nSMase inhibitor GW4869, decreased DA uptake in PC12 cells, although the application of FB1, a ceramide synthase inhibitor, did not affect DA uptake. Transfection of nSMase2 and CERK siRNA decreased DAT surface level in PC12 cells. These results suggested that SM-derived C1P affects cell surface levels of DAT.
科研通智能强力驱动
Strongly Powered by AbleSci AI