Metabolic Depletion of Sphingolipids Reduces Cell Surface Population of the Human Serotonin1A Receptor due to Impaired Trafficking

Sphingolipids and their metabolites are increasingly implicated in the pathogenesis of many metabolic and neurological diseases. It has been postulated that sphingolipids coalesce with cholesterol to form laterally segregated lipid domains that are involved in protein sorting and trafficking. In this work, we have explored the effect of metabolic depletion of sphingolipids on cell surface expression of the human serotonin(1A) receptor, a neurotransmitter G protein-coupled receptor. We used fumonisin B-1 (FB1), a fungal mycotoxin, to inhibit sphingolipid biosynthesis in HEK-293 cells stably expressing the human serotonin(1A) receptor. Our results obtained using flow cytometric analysis and confocal microscopic imaging show that the cell surface population of the serotonin(1A) receptor is reduced under sphingolipid-depleted condition. Western blot analysis confirmed that there was no significant difference in total cellular level of the serotonin(1A) receptor upon depletion of sphingolipids. Interestingly, the effect of FB1 on serotonin(1A) receptor population was reversed upon replenishment with sphingolipids. These results indicate that sphingolipid depletion does not alter total cellular receptor levels, but impairs serotonin(1A) receptor trafficking to the cellular plasma membrane. These results could provide mechanistic insights into the role of sphingolipids in modulation of neurotransmitter receptor signaling and trafficking in health and disease.

Automated summary

The depletion of sphingolipids has been shown to reduce the cell surface expression of the serotonin(1A) receptor, without altering the total cellular level of the receptor. This impairment in receptor trafficking was reversed upon replenishment with sphingolipids, indicating a role for sphingolipids in neurotransmitter receptor signaling and trafficking.