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  • Characterization of cadmium plasma membrane transport in gills of a mangrove crab Ucides cordatus.

Characterization of cadmium plasma membrane transport in gills of a mangrove crab Ucides cordatus.

Aquatic toxicology (Amsterdam, Netherlands) (2014-12-03)
P Ortega, M R Custódio, F P Zanotto
ABSTRACT

Membrane pathway for intracellular cadmium (Cd(2+)) accumulation is not fully elucidated in many organisms and has not been studied in crab gill cells. To characterize membrane Cd(2+) transport of anterior and posterior gill cells of Ucides cordatus, a hypo-hyper-regulating crab, a change in intracellular Cd(2+) concentration under various experimental conditions was examined by using FluoZin, a fluorescent probe. The membrane Cd(2+) transport was estimated by the augmentation of FluoZin fluorescence induced by extracellular application of CdCl2 and different inhibitors. Addition of extracellular calcium (Ca(2+)) to the cells affected little the fluorescence of FluoZin, confirming that Cd(2+) was the main ion increasing intracellular fluorescence. Ca(2+) channels blockers (nimodipine and verapamil) decreased Cd(2+) influx as well as vanadate, a Ca(2+)-ATPase blocker. Chelating intracellular Ca(2+) (BAPTA) decreased Cd(2+) influx in gill cells, while increasing intracellular Ca(2+) (caffeine) augmented Cd influx. Cd(2+) and ATP added at different temporal conditions were not effective at increasing intracellular Cd(2+) accumulation. Ouabain (Na(+)/K(+)-ATPase inhibitor) increased Cd(2+) influx probably through a change in intracellular Na and/or a change in cell membrane potential. Routes of Cd(2+) influx, a non-essential metal, through the gill cell plasma membrane of crabs are suggested.

MATERIALS
Product Number
Brand
Product Description

Supelco
Melting point standard 235-237°C, analytical standard
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Verapamil hydrochloride solution, 1.0 mg/mL in methanol (as free base), ampule of 1 mL, certified reference material, Cerilliant®
Caffeine for system suitability, European Pharmacopoeia (EP) Reference Standard
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Caffeine solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
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(±)-Verapamil hydrochloride, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, BioUltra, for molecular biology, ≥99.0% (T)
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Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, ≥97.0%
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Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, for molecular biology, ≥97.0%
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Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, BioXtra, ≥97 .0%
Supelco
Caffeine solution, analytical standard, 1.0 mg/mL in methanol
Sigma-Aldrich
(±)-Verapamil hydrochloride, ≥99% (titration), powder
Caffeine, European Pharmacopoeia (EP) Reference Standard
Cadmium, wire reel, 0.1m, diameter 0.70mm, as drawn, 99.99+%
Cadmium, wire reel, 0.1m, diameter 1.2mm, as drawn, 99.99+%
Cadmium, wire reel, 0.1m, diameter 2.0mm, hard, 99.99+%
Nimodipine, European Pharmacopoeia (EP) Reference Standard
Cadmium, rod, 150mm, diameter 10.0mm, as drawn, 99.99+%
Cadmium, foil, light tested, 100x100mm, thickness 0.05mm, as rolled, 99.99+%
Cadmium, rod, 200mm, diameter 10.0mm, 99.999%
Cadmium, tube, 500mm, outside diameter 2.29mm, inside diameter 1.27mm, wall thickness 0.51mm, 99.95+%
Cadmium, foil, not light tested, 50x50mm, thickness 0.002mm, 99.7+%
Cadmium, foil, light tested, 50x50mm, thickness 0.05mm, as rolled, 99.99+%
Cadmium, tube, 200mm, outside diameter 2.29mm, inside diameter 1.27mm, wall thickness 0.51mm, 99.95+%
Cadmium, foil, not light tested, 100x100mm, thickness 0.015mm, 99.7+%
Cadmium, wire reel, 0.1m, diameter 0.25mm, hard, 99.99+%
Cadmium, foil, light tested, 25x25mm, thickness 0.015mm, 99.7+%
Cadmium, foil, not light tested, 100x100mm, thickness 0.01mm, 99.7+%
Cadmium, foil, not light tested, 100x100mm, thickness 0.005mm, 99.7+%
Cadmium, rod, 25mm, diameter 40mm, 99.9%
Cadmium, foil, not light tested, 25x25mm, thickness 0.015mm, 99.7+%