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Some characteristics of membrane Cd2+ transport in rat thymocytes: an analysis using Fluo-3

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Abstract

Although cadmium-induced apoptosis of lymphocytes is one of common features in the immunotoxicity of cadmium, the membrane pathway for intracellular cadmium accumulation is not fully elucidated. To characterize membrane Cd2+ transport of rat thymocytes, the change in intracellular Cd2+ concentration under various conditions was examined by the use of Fluo-3, a fluorescent probe for monitoring the change in intracellular concentration of divalent metal cations. The membrane Cd2+ transport was estimated by the augmentation of Fluo-3 fluorescence induced by bath application of CdCl2. Lowering temperature strongly suppressed the augmentation of Fluo-3 fluorescence by CdCl2, suggesting that the metabolic process can be involved in membrane Cd2+ transport. External acidification (decreasing pH) and membrane depolarization by adding KCl attenuated the augmentation, indicating the requirement of electrochemical driving force for membrane Cd2+ transport into the cells. Bath application of CaCl2 and ZnCl2 equally decreased the augmentation, suggesting their competition with Cd2+ at the membrane transport. The augmentation by CdCl2 was lesser in the cells treated with N-ethylmaleimide inducing chemical depletion of cellular thiols. The result suggests the contribution of sulfhydryl groups to membrane Cd2+ transport. Taken together, it is suggested that the cells possess a temperature-sensitive membrane Cd2+ pathway, driven by electrochemical gradient of Cd2+ and transmembrane potential, with competitive binding site. Based on the characteristics described above, it is unlikely that the membrane Cd2+ transport in rat thymocytes is attributed to a single transport system although it has characteristics that are similar to those of divalent cation transporter 1.

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Acknowledgment

We thank the reviewer for the comments concerning some additional experiments.

Conflict of interest

We have no conflicts of interest on this study. This study was carried out by the institutional expenditure (University of Tokushima).

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Authors and Affiliations

  1. Laboratories of Cellular Signaling, Environmental Biology, and Organometallic Chemistry, Faculty of Integrated Arts and Sciences, The University of Tokushima, 1-1 Minami-Jyosanjima, Tokushima, 770-8502, Japan

    Takuya Kawanai, Masahiro Fujinaga, Kazuki Koizumi, Isao Kurotani, Erika Hashimoto, Masaya Satoh, Shoji Imai, Norikazu Miyoshi & Yasuo Oyama

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  1. Takuya Kawanai
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  2. Masahiro Fujinaga
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  3. Kazuki Koizumi
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  4. Isao Kurotani
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  5. Erika Hashimoto
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  7. Shoji Imai
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  8. Norikazu Miyoshi
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  9. Yasuo Oyama
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Correspondence to Yasuo Oyama.

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Kawanai, T., Fujinaga, M., Koizumi, K. et al. Some characteristics of membrane Cd2+ transport in rat thymocytes: an analysis using Fluo-3. Biometals 24, 903–914 (2011). https://doi.org/10.1007/s10534-011-9444-3

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  • DOI: https://doi.org/10.1007/s10534-011-9444-3

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