Impact of environmental salinity on the MAPK-NFAT5 pathway in Trachemys scripta elegans and its role in osmoregulaton

Globally, sea level rise (SLR) leads to salinization of coastal freshwater, in where organisms might be affected and in turn promote osmoregulation and adaptation in response to freshwater salinization. Trachemys scripta elegans a freshwater turtle species, exhibits remarkable tolerance to varying salinity environments, yet the underlying regulatory mechanisms remain poorly understood. This study aimed to elucidate the molecular mechanisms of osmoregulation in this species based on previous RNA-seq data. Our findings revealed that when exposed to 5 PSU (5 ‰) and 15 PSU (15 ‰) salinities, the turtles exhibited increased concentrations of ions (Na⁺, K⁺) and urea in urine, along with elevated osmotic pressures in both plasma and urine. Additionally, the protein levels of aquaporins (AQPs) and transporters of ions and organic osmolytes in the kidney were upregulated in saline water. Notably, the transcriptional level of the hypertonic regulator NFAT5 was significantly elevated, accompanied by an increase in phosphorylated NFAT5 levels in the nucleus of renal tubular epithelial cells. Furthermore, we observed upregulated phosphorylated levels of MAPKs in saline water. The use of MAPK inhibitors effectively blocked the transcription of NFAT5 and osmoregulatory target genes. Collectively, these results suggest that T. scripta elegans activates the MAPK-NFAT5 signaling pathway to modulate osmotic pressure in adaptation to saline water environments. Our study provides valuable insights into the osmoregulatory responses of aquatic organisms to saline environments and aids in understanding the adaptability of organisms inhabiting coastal areas facing rising sea levels.