Abstract
Sea lamprey (Petromyzon marinus) begin life as filter-feeding larvae (ammocoetes) before undergoing a complex metamorphosis into parasitic juveniles, which migrate to the sea where they feed on the blood of large-bodied fishes. The greater protein intake during this phase results in marked increases in the production of nitrogenous wastes (N-waste), which are excreted primarily via the gills. However, it is unknown how gill structure and function change during metamorphosis and how it is related to modes of ammonia excretion, nor do we have a good understanding of how the sea lamprey’s transition from fresh water (FW) to sea water (SW) affects patterns and mechanisms of N-waste excretion in relation to ionoregulation. Using immunohistochemistry, we related changes in the gill structure of larval, metamorphosing, and juvenile sea lampreys to their patterns of ammonia excretion (Jamm) and urea excretion (Jurea) in FW, and following FW to artificial seawater (ASW) transfer. Rates of Jamm and Jurea were low in larval sea lamprey and increased in feeding juvenile, parasitic sea lamprey. In freshwater-dwelling ammocoetes, immunohistochemical analysis revealed that Rhesus glycoprotein C-like protein (Rhcg-like) was diffusely distributed on the lamellar epithelium, but following metamorphosis, Rhcg-like protein was restricted to SW mitochondrion-rich cells (MRCs; ionocytes) between the gill lamellae. Notably, these interlamellar Rhcg-like proteins co-localized with Na+/K+-ATPase (NKA), which increased in expression and activity by almost tenfold during metamorphosis. The distribution of V-type H+-ATPase (V-ATPase) on the lamellae decreased following metamorphosis, indicating it may have a more important role in acid–base regulation and Na+ uptake in FW, compared to SW. We conclude that the re-organization of the sea lamprey gill during metamorphosis not only plays a critical role in allowing them to cope with greater salinity following the FW–SW transition, but that it simultaneously reflects fundamental changes in methods used to excrete ammonia.
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Acknowledgements
We wish to acknowledge Dr. S. Hirose who generously provided us with the Rhcg1 antibodies used for our immunohistochemistry and western blot analyses. We also thank Dr. Margaret Docker (Biological Sciences, University of Manitoba) who graciously provided her time and expertise to help us identify the stages of sea lamprey metamorphosis. We also thank Dr. Jeremiah Smith, who provided us with guidance and access to the unpublished lamprey transcriptome and genome data. Finally, we are grateful to Laura Tessier, Phillip Pham-Ho, Josh Sutherby, Heather Bauman and Vedya Ramsaran who assisted with the collection of larval sea lamprey, and tissue collections. This work was funded by a National Sciences and Engineering Research Council (NSERC) of Canada Discovery Grant awarded to MPW.
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MPW and JMW: conceived the study, developed the experimental protocols and provided significant editorial guidance for the drafting of the manuscript; JS: carried out the animal collection and care, coordinated and performed the experiments, statistical analysis and drafted the manuscript as part of her M.Sc. studies. All authors provided final approval for publication.
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Sunga, J., Wilson, J.M. & Wilkie, M.P. Functional re-organization of the gills of metamorphosing sea lamprey (Petromyzon marinus): preparation for a blood diet and the freshwater to seawater transition. J Comp Physiol B 190, 701–715 (2020). https://doi.org/10.1007/s00360-020-01305-1
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DOI: https://doi.org/10.1007/s00360-020-01305-1