Single molecule real-time sequencing revealing novel insights on the response to estrogen and androgen exposure in freshwater snails.

ABSTRACT

The molecular mode of action underpinning the response of mollusks exposure to endocrine disrupting chemicals (EDCs) remains unclear due to a lack of available information regarding their genome. Single molecule real-time (SMRT) sequencing makes it possible to reveal molecular mechanisms by direct sequencing of full-length transcripts. In the present study, the transcriptome profile of the freshwater snail Parafossarulus striatulus after exposure to 17ß-estradiol (E2) or 17α-methyltestosterone (MT) was evaluated using SMRT sequencing strategy. In total, 216,598 non-redundant and full-length gene isoforms were generated and 106,266 isoforms were predicted with a complete open reading frame (ORF). Moreover, 60.36% of the isoforms were matched to known proteins in at least one of six databases. Differential gene expression analyses showed significantly different patterns in paired samples with different treatments. The expression levels of several membrane receptor isoforms of P. striatulus including dopamine receptor (DR), FMRFamide receptor (FMRFaR), neuropeptide Y receptor (NYR) and neuropeptide FF receptor (NFFR), but not estrogen receptor (ER) or estrogen-related receptor (ERR), were significantly affected by E2 and MT. These findings suggest that activation of membrane receptors, as well as other signaling pathways, might be critical for mediating the effects of endocrine disruption in mollusks. The transcriptome information obtained from the SMRT sequencing provides a significant contribution to the investigation of the molecular mode of action of endocrine disrupting chemicals on P. striatulus.