Disturbance in Na+ regulation in cells rich in mitochondria isolated from gills of the yellow clam Mesodesma mactroides exposed to copper under different osmotic conditions

Cells rich in mitochondria were isolated from gills of the seawater clam Mesodesma mactroides, incubated in isosmotic saline solution (840 mOsmol/kg H₂O), and exposed (3 h) to environmentally realistic Cu concentrations (nominally: 0, 5, 9 and 20 μg/L). In cells exposed to 20 μg Cu/L, Cu accumulation, Na⁺ content reduction and carbonic anhydrase (CA) activity inhibition were observed, without significant changes in cell viability and Na⁺,K⁺-ATPase (NKA) activity. In the absence of Cu, cell viability and Cu content were reduced in hyposmotic media respect with the control, without changes in Na⁺ content and enzyme (CA and NKA) activities. In the presence of 5 or 9 μg/L Cu, cell Cu content was increased, especially at 670 mOsmol/kg H₂O. Cell Na⁺ content and NKA activity were reduced after exposure to 20 μg/L Cu at 670 mOsmol/kg H₂O. In turn, CA activity was dependent on Cu concentration, being significantly reduced in cells exposed to 9 and 20 μg/L Cu in both hyposmotic conditions. These findings indicate that Cu also negatively affects Na⁺ regulation in gill cells of the seawater clam M. mactroides, with Cu toxicity increasing at hyposmotic conditions. Also, they indicate that physiology is more important than water chemistry in predicting Cu toxicity in environments of changing salinity, pointing out CA activity as a potential biomarker of Cu exposure.