Alanine, proline and urea are major organic osmolytes in the snailTheodoxus fluviatilisunder hyperosmotic stress

Hyperosmotic stress may result in osmotic volume loss from the body to the environment in animals that cannot control the water permeability of their integument. Euryhaline animals (which have a wide tolerance range of environmental salinities) have generally evolved the ability to counteract cell volume shrinkage by accumulating inorganic and organic osmolytes within their cells to balance internal and external osmolalities. Molluscs use very different combinations of amino acids and amino acid derivatives to achieve this goal. Theodoxus fluviatilis is a neritid gastropod that is distributed not only in limnic habitats in Europe but also in brackish waters (e.g. along the shoreline of the Baltic Sea). Animals from brackish sites survive better in high salinities than animals from freshwater locations. The results of the present study indicate that these differences in salinity tolerance cannot be explained by differences in the general ability to accumulate amino acids as organic osmolytes. Although there may be differences in the metabolic pathways involved in osmolyte accumulation in foot muscle tissue, the two groups of animals accumulate amino acid mixtures equally well when stepwise acclimated to their respective maximum tolerable salinity for extended periods. Among these amino acids, alanine and proline, as well as the osmolyte urea, hold a special importance for cell volume preservation in T. fluviatilis under hyperosmotic stress. It is possible that the accumulation of various amino acids during hyperosmotic stress occurs via hydrolysis of storage proteins, while alanine and proline are probably newly synthesised under conditions of hyperosmotic stress in the animals.