Instituto de Biociências, Departamento de Fisiologia, Universidade de São Paulo, Rua do Matão, Travessa 14, #101, São Paulo 05508-900, SP, Brazil.
Instituto de Biociências, Departamento de Fisiologia, Universidade de São Paulo, Rua do Matão, Travessa 14, #101, São Paulo 05508-900, SP, Brazil; Departamento de Biofísica, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Três de Maio 100, São Paulo 04044-020, Brazil.
Aquat Toxicol. 2014 Dec;157:21-9. doi: 10.1016/j.aquatox.2014.09.006. Epub 2014 Oct 2.
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.
在许多生物中,细胞内镉(Cd(2+))积累的膜途径尚未完全阐明,在蟹鳃细胞中也尚未进行研究。为了描述 Ucides cordatus(一种低-高调节蟹)前、后鳃细胞的膜 Cd(2+)转运,我们使用荧光探针 FluoZin 检测了各种实验条件下细胞内 Cd(2+)浓度的变化。通过测量细胞外应用 CdCl2 和不同抑制剂引起的 FluoZin 荧光增强来估算膜 Cd(2+)转运。向细胞中添加细胞外钙(Ca(2+))对 FluoZin 的荧光影响很小,证实 Cd(2+)是增加细胞内荧光的主要离子。Ca(2+)通道阻滞剂(尼莫地平、维拉帕米)和钒酸盐(一种 Ca(2+)-ATP 酶抑制剂)均降低了 Cd(2+)内流。螯合细胞内 Ca(2+)(BAPTA)降低了鳃细胞中的 Cd(2+)内流,而增加细胞内 Ca(2+)(咖啡因)则增强了 Cd 内流。在不同的时间条件下添加 Cd(2+)和 ATP 对细胞内 Cd(2+)积累没有效果。哇巴因(Na(+)/K(+)-ATP 酶抑制剂)增加了 Cd(2+)内流,可能是通过改变细胞内 Na 和/或细胞膜电位。这些结果提示了非必需金属 Cd(2+)通过蟹鳃细胞膜的内流途径。