Integrated application of multi-omics approach and biochemical assays provides insights into physiological responses to saline-alkaline stress in the gills of crucian carp (Carassius auratus)

Abstract

Given the decline of freshwater resources in recent years, the accessible space for freshwater aquaculture is rapidly shrinking, and aquaculture in saline-alkaline water has become a critical approach to meet the rising demand. However, the molecular mechanism behind the adverse effects of saline-alkaline water on fish and the regulatory mechanism in fish tolerance remains unclear. Here, adult crucian carp (Carassius auratus) were exposed to 60 mmol/L NaHCO₃ for 30 days. It was observed that long-term carbonate alkalinity (CA) exposure not only caused gill oxidative stress but also changed the levels of several physiological parameters associated with ammonia transport, including blood ammonia, urea nitrogen (BUN), glutamine (Gln), and glutamine synthetase (GS). According to the metabolomics study, differential metabolites (DMs) engaged in various metabolic pathways, such as glycerophospholipid metabolism, sphingolipid metabolism, and arachidonic acid metabolism. In addition, transcriptomics data showed that differentially expressed genes (DEGs) were closely related to ammonia transport, apoptosis, and immunological response. In general, comprehensive multi-omics and biochemical analysis revealed that crucian carp might adopt Rh glycoprotein as a carrier to mediate ammonia transport and increase glutamine and urea synthesis under long-term high saline-alkaline stress to mitigate the adverse effects of blocked ammonia excretion. Simultaneously, saline-alkaline stress caused the destruction of the antioxidant system and the disorder of lipid metabolism in the crucian carp gills, which induced apoptosis and immunological response. To our knowledge, this is the first study to investigate fish's molecular and metabolic mechanisms under saline-alkaline stress using integrated metabolomics, transcriptomics, and biochemical assays. Overall, the results of this study provided new insights into the molecular mechanism behind the adverse effects of saline-alkaline water on fish and the regulatory mechanism in fish tolerance.

Keywords: Crucian carp; Gill; Metabolomics; Saline-alkaline stress; Transcriptomics.