The functional role of a serotonin transporter in Schistosoma mansoni elucidated through immunolocalization and RNA interference (RNAi)

Serotonin is an important neurotransmitter in both vertebrates and invertebrates. In the parasitic flatworm, Schistosoma mansoni, serotonin stimulates worm movement and potentiates muscle contraction. A specific serotonin transporter (SmSERT) was previously cloned from S. mansoni and characterized in vitro. Here we conduct a first investigation of the native protein in the worm so as to elucidate the biological role of SmSERT and to assess its drug targeting potential. Confocal immunofluorescence studies using specific antibodies determined that SmSERT is expressed predominantly in the nervous system both in adult worms and larvae (schistosomula). SmSERT immunoreactivity was detected in the main nerve cords of the central nervous system and the peripheral nerve plexus of the body wall in adult males and females, in apparent nerve endings of the male tubercles and possibly the male tegument. In the larvae, SmSERT localized mainly to the peripheral nerve plexus of the body wall. Co-localization experiments showed that the pattern of SmSERT expression coincides with that of serotonin itself, suggesting that SmSERT is present in serotonergic neurons. To test whether SmSERT is involved in the motor effects of serotonin, we treated S. mansoni schistosomula with SmSERT blockers or SmSERT-specific short-interfering RNAs (siRNAs) and then recorded larval motility, using a quantitative imaging assay. In both cases, the treatment produced a strongly hyperactive phenotype, corresponding to a ∼3-fold increase in larval motility, roughly the same effect as treatment with an excess of exogenous serotonin. The siRNA effect correlated with a ≈50% decrease in expression of the SmSERT when tested by real-time qPCR. To test if SmSERT mediates transport of exogenous serotonin across the tegument, uptake assays were also performed in intact schistosomula treated with SmSERT siRNAs or an irrelevant siRNA. We found a significant but modest decrease (∼25%) in serotonin uptake in the siRNA-suppressed larvae when compared to the negative controls. These results suggest that the SmSERT's function is primarily neuromuscular and may also play a secondary role in the uptake of exogenous (host-derived) serotonin.

72Highlights► Serotonin is an important neurotransmitter and modulator of motility in schistosomes. ► A serotonin transporter (SmSERT) was identified in serotonergic neurons of Schistosoma mansoni. ► RNAi targeting SmSERT caused significant hyperactivity in cultured S. mansoni schistosomula. ► Schistosomula treated with SmSERT inhibitors also exhibited hyperactivity. ► We propose a model by which SmSERT regulates serotonin signaling and parasite motility.