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The (Na+,K+)-ATPase in osmoregulatory capacity in crab Ucides cordatus: biochemical and molecular view

Grant number: 13/24252-9
Support type:Scholarships in Brazil - Post-Doctorate
Effective date (Start): March 01, 2014
Effective date (End): February 28, 2017
Field of knowledge:Biological Sciences - Biochemistry
Principal Investigator:Francisco de Assis Leone
Grantee:Malson Neilson de Lucena
Home Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

This postdoctoral project is a continuation our line of research that has been supported by FAPESP in which we investigate the gill (Na+,K+)-ATPase in crustaceans as molecular marker of adaptation to environments of different salinities. Freshwater species face the challenge to develop physiological adaptations under hyposmotic conditions. In hyperosmoregulating animals, the gills act as a selective interface that actively absorbs Na+ and Cl- ions from the dilute external media to the hemolymph; the (Na+, K+)-ATPase and V-ATPase are key molecules in this process with other transporters being present in the basolateral and apical membranes. Ucides cordatus is naturally exposed to wide fluctuation in salinity, from 2 to 33 0, mainly resulting from tidal flow in mangroves. These salinity changes (either increase or decrease) can affect the expression/activities of the antioxidant enzyme system, the ion transporting enzymes, and oxygen consumption. In this project we will study the activities of the above enzymes, aiming to understand the mechanisms involved in the biochemical, physiological and molecular adaptations during the invasion of the freshwater environment by crustaceans, and in ammonium excretion, using the mangrove crab U. cordatus. To characterize osmoregulatory capacity in U. cordatus, crabs will be acclimated to 2 0, 26 0 and 34 0 salinity for ten days. For each acclimation condition the following steps will be performed: 1) evaluate the (Na+,K+)-ATPase, V-ATPase, Ca2+-ATPase and carbonic anhydrase activities; 2) examine the modulation of each activity by their respective(s) effector(s); 3) effect of specific inhibitors; 4) ammonium excretion; 5) phosphoenzyme formation; 6) effect of FXYD on (Na+,K+)-ATPase activity; 7) ionic concentration and hemolymph osmolality; 8) mRNA expression of ± and B subunits of (Na+,K+)-ATPase and V-ATPase; 9) oxygen consumption; 10) estimate activity of oxidative stress enzymes; 11) evaluate the effect of both K+ and NH4+ on (Na+,K+)-ATPase activity and phosphoenzyme formation; 12) evaluate (Na+,K+)-ATPase activity after acclimation to different ammonium sulfate concentrations; 13) immunohistochemical localization of (Na+,K+)-ATPase and V-ATPase.

Scientific publications (8)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
LUCENA, MALSON N.; GARCON, DANIELA P.; FONTES, CARLOS F. L.; FABRI, LEONARDO M.; MORAES, CINTYA M.; MCNAMARA, JOHN C.; LEONE, FRANCISCO A. Dopamine binding directly up-regulates (Na+, K+)-ATPase activity in the gills of the freshwater shrimp Macrobrachium amazonicum. COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY A-MOLECULAR & INTEGRATIVE PHYSIOLOGY, v. 233, p. 39-47, JUL 2019. Web of Science Citations: 0.
FALEIROS, ROGERIO O.; GARCON, DANIELA P.; LUCENA, MALSON N.; MCNAMARA, JOHN C.; LEONE, FRANCISCO A. Short- and long-term salinity challenge, osmoregulatory ability, and (Na+, K+)-ATPase kinetics and alpha-subunit mRNA expression in the gills of the thinstripe hermit crab Clibanarius symmetricus (Anomura, Diogenidae). COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY A-MOLECULAR & INTEGRATIVE PHYSIOLOGY, v. 225, p. 16-25, NOV 2018. Web of Science Citations: 3.
FARIAS, DANIEL L.; LUCENA, MALSON N.; GARCON, DANIELA P.; MANTELATTO, FERNANDO L.; MCNAMARA, JOHN C.; LEONE, FRANCISCO A. A Kinetic Characterization of the Gill (Na+, K+)-ATPase from the Semi-terrestrial Mangrove Crab Cardisoma guanhumi Latreille, 1825 (Decapoda, Brachyura). Journal of Membrane Biology, v. 250, n. 5, p. 517-534, OCT 2017. Web of Science Citations: 1.
ANTUNES, CLAUDIA DOI; LUCENA, MALSON NEILSON; GARCON, DANIELA PEREIRA; LEONE, FRANCISCO ASSIS; MCNAMARA, JOHN CAMPBELL. Low salinity-induced alterations in epithelial ultrastructure, Na+/K+-ATPase immunolocalization and enzyme kinetic characteristics in the gills of the thinstripe hermit crab, Clibanarius vittatus (Anomura, Diogenidae). JOURNAL OF EXPERIMENTAL ZOOLOGY PART A-ECOLOGICAL AND INTEGRATIVE PHYSIOLOGY, v. 327, n. 6, p. 380-397, JUL 2017. Web of Science Citations: 2.
LUCENA, MALSON NEILSON; GARCON, DANIELA PEREIRA; LEITE FONTES, CARLOS FREDERICO; MCNAMARA, JOHN CAMPBELL; LEONE, FRANCISCO ASSIS. Polyamines regulate phosphorylation-dephosphorylation kinetics in a crustacean gill (Na+, K+)-ATPase. Molecular and Cellular Biochemistry, v. 429, n. 1-2, p. 187-198, MAY 2017. Web of Science Citations: 4.
LUCENA, MALSON N.; MCNAMARA, JOHN C.; LEONE, FRANCISCO A. Gill (Na+, K+)-ATPase from the Amazon River shrimp, Macrobrachium amazonicum (Decapoda, Palaemonidae): effect of exogenous biogenic amines on enzyme activity in juveniles and adults. Hydrobiologia, v. 789, n. 1, p. 59-76, MAR 2017. Web of Science Citations: 5.
PINTO, MARCELO R.; LUCENA, MALSON N.; FALEIROS, ROGERIO OLIVEIRA; ALMEIDA, EDUARDO ALVES; MCNAMARA, JOHN C.; LEONE, FRANCISCO A. Effects of ammonia stress in the Amazon river shrimp Macrobrachium amazonicum (Decapoda, Palaemonidae). AQUATIC TOXICOLOGY, v. 170, p. 13-23, JAN 2016. Web of Science Citations: 20.
LEONE, FRANCISCO A.; GARCON, DANIELA P.; LUCENA, MALSON N.; FALEIROS, ROGERIO O.; AZEVEDO, SERGIO V.; PINTO, MARCELO R.; MCNAMARA, JOHN C. Gill-specific (Na+, K+)-ATPase activity and alpha-subunit mRNA expression during low-salinity acclimation of the ornate blue crab Callinectes ornatus (Decapoda, Brachyura). COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, v. 186, p. 59-67, AUG 2015. Web of Science Citations: 13.

Please report errors in scientific publications list by writing to: cdi@fapesp.br.