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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

Sigma-Aldrich
Caffeine, anhydrous, 99%, FCC, FG
Supelco
Caffeine, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Caffeine melting point standard, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Caffeine, anhydrous, tested according to Ph. Eur.
Sigma-Aldrich
Caffeine, powder, ReagentPlus®
Sigma-Aldrich
Caffeine, Sigma Reference Standard, vial of 250 mg
USP
Verapamil hydrochloride, United States Pharmacopeia (USP) Reference Standard
Supelco
Caffeine, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Caffeine, European Pharmacopoeia (EP) Reference Standard
Nimodipine, European Pharmacopoeia (EP) Reference Standard
Verapamil hydrochloride, European Pharmacopoeia (EP) Reference Standard
USP
Caffeine, United States Pharmacopeia (USP) Reference Standard
Supelco
(±)-Verapamil hydrochloride, Pharmaceutical Secondary Standard; Certified Reference Material
Caffeine for system suitability, European Pharmacopoeia (EP) Reference Standard