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Ammocoetes of the anadromous sea lamprey Petromyzon marinus L. spend many years in freshwater before metamorphosing and migrating to sea. Metamorphosis involves the radical transformation from a substrate-dwelling, filter feeder into a free-swimming, parasitic feeder. In the present work we examined osmoregulatory differences between ammocoetes and transformers (metamorphic juveniles), and the effects of salinity acclimation. We measured the expression of key ion-transporting proteins [Na(+)/K(+)-ATPase, vacuolar (V)-type H(+)-ATPase and carbonic anhydrase (CA)] as well as a number of relevant blood parameters (hematocrit, [Na(+)] and [Cl(-)]). In addition, immunofluorescence microscopy was used to identify and characterize the distributions of Na(+)/K(+)-ATPase, V-type H(+)-ATPase and CA immunoreactive cells in the gill. Ammocoetes did not survive in the experiments with salinities greater than 10 per thousand, whereas survival in high salinity (&gt; or =25-35 per thousand) increased with increased degree of metamorphosis in transformers. Plasma [Na(+)] and [Cl(-)] of ammocoetes in freshwater was lower than transformers and increased markedly at 10 per thousand. In transformers, plasma ions increased only at high salinity (&gt;25 per thousand). Branchial Na(+)/K(+)-ATPase levels were approximately tenfold higher in transformers compared to ammocoetes and salinity did not affect expression in either group. However, branchial H(+)-ATPase expression showed a negative correlation with salinity in both groups. Na(+)/K(+)-ATPase immunoreactivity was strongest in transformers and associated with clusters of cells in the interlamellar spaces. H(+)-ATPase (B subunit) immunoreactivity was localized to epithelial cells not expressing high Na(+)/K(+)-ATPase immunoreactivity and having a similar tissue distribution as carbonic anhydrase. The results indicate that branchial Na(+)/K(+)-ATPase and salinity tolerance increase in metamorphosing lampreys, and that branchial H(+)-ATPase is downregulated by salinity.

Ionoregulatory changes during metamorphosis and salinity exposure of juvenile sea lamprey (Petromyzon marinusL.)