The Ins and Outs of Urea: Identification of Putative DUR3-like Urea Transporters in the Oligohaline Nerite Snail Theodoxus fluviatilis and Their Expression under Changing Salinities

Abstract

Theodoxus fluviatilis (Linnaeus, 1758) (Gastropoda: Neritidae) is an oligohaline aquatic gastropod that inhabits most of Europe and adjacent areas of Asia. Two different ecotypes can be distinguished: One in freshwater (FW) and another along the Baltic Sea coast in brackish water habitats (BW). Individuals of either ecotype use free amino acids and urea as organic osmolytes to adjust body fluid osmolality to the external medium; however, the BW ecotype is able to accumulate them in larger quantities. The use of urea as an organic osmolyte in aquatic gastropods such as T. fluviatilis has only recently been initially described and raised the question of how urea transport between body fluids and the environment is balanced. Upon examining transcriptome and preliminary genome sequence data of T. fluviatilis, we identified putative homologues of DUR3 genes, which code for urea transporters (UTs) in other organisms. In this study, we provide evidence for the presence of four different subtypes of DUR3-like UTs that belong to two distinct families. Two of the UT subtypes were subject to qRT-PCR analyses to investigate differences in mRNA expression during the acclimation of individuals of both ecotypes to different salinities. Our results indicate that only BW animals regulate DUR3 gene expression in the context of osmoregulation.

A draft genome of the neritid snail Theodoxus fluviatilis

“…The whole procedure of RNA extraction, cDNA library construction, and Mi-seq sequencing was already described in great detail in Knobloch et al (2023) . Briefly, a pool of foot muscle preparations of 24 individuals that were collected at freshwater and brackish water locations in Northern Germany and exposed to different salinities under laboratory conditions were used to extract total RNA.…”