Aquaculture Reports (Apr 2024)
Molecular and physiological responses in the ammonia transport pathways in clam Cyclina sinensis exposed to chronic ammonia nitrogen
Abstract
The clam Cyclina sinensis is characterized by good ammonia resistance. With the purpose of gaining knowledge about its ammonia resistance, we assessed the mRNA expression levels of Rhesus glucoprotein b (CsRhbg), the activities of key enzymes in the ammonia transport pathways (glutamine synthetase, GS; glutamic acid dehydrogenase, GDH) in the gill and hepatopancreas and the gill histopathological section (HE staining) of the clam exposed to total ammonia nitrogen (TAN) (6.58 mg/L) for 768 h (32d). Results revealed that the full-length cDNA sequence of CsRhbg was 1777 bp in length. The CsRhbg was highly conserved in the same genus, and the protein encoded by CsRhbg contains 10 transmembrane structures (TMHs) and one signal peptide. Exposed to ammonia nitrogen, the expression level of CsRhbg mRNA climbed sharply at 3 h post-exposure, and then it decreased and fluctuated from 192 h to 768 h. The activity of GSase in the clam exposed to ammonia nitrogen increased in the first 24 h, and reached the highest value at 96 h, and then fluctuated. The activity of GDH rose sharply and reached the highest value at 3 h, and then fluctuated. With the increase of exposure time, histological alterations in the gill became more and more clear in the disappearance of the epithelial cell boundaries. The results suggest that exposure to ammonia nitrogen can activate CsRhbg gene expression, and elevate the GSase and GDH activity in the gill of the clam as the ammonia detoxification strategy to reduce the harm of ammonia. However, prolonged exposure to ammonia nitrogen can result in the down-regulation of CsRhbg gene, dysfunction of key enzymes in the ammonia transport pathways, and damage to the structure of epithelial cells or the cellular structure in the gill of the clam. Therefore, this work will help to reveal the ammonia detoxification strategy in bivalves exposed to chronic ammonia nitrogen.