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Role of (Na,K)-ATPase in osmoregulation and excretion of ammonium ions in euryhaline crustaceans: identification and influence of FXYD2 on the regulation of gill (Na,K)-ATPase of the crab Callinectes danae

Grant number: 10/17534-0
Support type:Regular Research Grants
Duration: January 01, 2011 - June 30, 2013
Field of knowledge:Biological Sciences - Biochemistry
Principal Investigator:Francisco de Assis Leone
Grantee:Francisco de Assis Leone
Home Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Assoc. researchers:Arthur Henrique Cavalcante de Oliveira ; Daniela Pereira Garçon ; Fernando Luis Medina Mantelatto ; John Campbell McNamara ; Klaus Hartmann Hartfelder ; Wagner Cotroni Valenti

Abstract

This project is a continuation our line of a long studying program research that has been supported by FAPESP with the aim of the systematic study of the structural properties and kinetics of Na,K-ATPase from gill tissue of different species of crabs and shrimps in order to establish a model for the adaptation of crustaceans to environments of different salinities. The maintained osmolarity of the hemolymph of crustaceans is attributed to the Na,K-ATPase and the mechanism of adaptation suggests that the responses of crustaceans must involve a rapid activation of the transport of ions that must be primary to any other physiological responses of these animals. The current project aims to investigate the cellular signaling linked to the regulation of gill Na,K-ATPase and its direct consequences on the uptake of NaCl during acclimation to hyposaline environments and marine environments. The following studies will be undertaken: 1) correlate the process of acclimation of Callinectes danae with differential expression of FXYD2 (Na,K-ATPase g subunit); 2) study the effect of FXYD2 phosphorylation of Na,K-ATPase of C. danae by protein kinase A; 3) cloning and sequencing of FXYD2 of Na,K-ATPase from C. danae gills; 4) study the effect of polyamines in different saline environments, related to the protein kinases A and C that are activated during osmotic stress, using cultured gill cells from C. danae; 5) elaboration of a protocol for purification of Na,K-ATPase of C. danae in order to obtain an enriched fraction of the enzyme for the identification during proteolytic activation, and subsequently linked to the conformation site binding NH+4; 6) identify the role that other endogenous regulators (dopamine, octopamine) in Na,K-ATPase regulation in cultured gill cells; 7) establish the sequence of aminoacids of Na,K-ATPase in order to identify aminoacid residues involved in cation binding; 8) correlate the FXYD2 with protein kinases stimulated by polyamines and its final action on the activity of Na,K-ATPase in different saline media, using cultured gill cells of C. danae. (AU)

Scientific publications (10)
(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)
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: 4.
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.
LEONE, FRANCISCO ASSIS; LUCENA, MALSON NEILSON; REZENDE, LUCIANA AUGUSTO; GARCON, DANIELA PEREIRA; PINTO, MARCELO RODRIGUES; MANTELATTO, FERNANDO LUIS; MCNAMARA, JOHN CAMPBELL. A Kinetic Characterization of (Na+, K+)-ATPase Activity in the Gills of the Pelagic Seabob Shrimp Xiphopenaeus kroyeri (Decapoda, Penaeidae). Journal of Membrane Biology, v. 248, n. 2, p. 257-272, APR 2015. Web of Science Citations: 6.
LUCENA, MALSON N.; PINTO, MARCELO R.; GARCON, DANIELA P.; MCNAMARA, JOHN C.; LEONE, FRANCISCO A. A kinetic characterization of the gill V(H+)-ATPase in juvenile and adult Macrobrachium amazonicum, a diadromous palaemonid shrimp. COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, v. 181, p. 15-25, MAR 2015. Web of Science Citations: 10.
LEONE, FRANCISCO A.; BEZERRA, THAIS M. S.; GARCON, DANIELA P.; LUCENA, MALSON N.; PINTO, MARCELO R.; FONTES, CARLOS F. L.; MCNAMARA, JOHN C. Modulation By K+ Plus NH4+ of Microsomal (Na+, K+)ATPase Activity in Selected Ontogenetic Stages of the Diadromous River Shrimp Macrobrachium amazonicum (Decapoda, Palaemonidae). PLoS One, v. 9, n. 2 FEB 21 2014. Web of Science Citations: 12.

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