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Methyl-β-cyclodextrin reversibly alters the gating of lipid rafts-associated Kv1.3 channels in Jurkat T lymphocytes

  • Cell and Molecular Physiology
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Abstract

The voltage-dependent Kv1.3 potassium channels mediate a variety of physiological functions in human T lymphocytes. These channels, along with their multiple regulatory components, are localized in cholesterol-enriched microdomains of plasma membrane (lipid rafts). In this study, patch-clamp technique was applied to explore the impact of the lipid-raft integrity on the Kv1.3 channel functional characteristics. T lymphoma Jurkat cells were treated for 1 h with cholesterol-binding oligosaccharide methyl-β-cyclodextrin (MβCD) in 1 or 2 mM concentration, resulting in depletion of cholesterol by 63 ± 5 or 75 ± 4%, respectively. Treatment with 2 mM MβCD did not affect the cells viability but retarded the cell proliferation. The latter treatment caused a depolarizing shift of the Kv1.3 channel activation and inactivation by 11 and 6 mV, respectively, and more than twofold decrease in the steady-state activity at depolarizing potentials. Altogether, these changes underlie the depolarization of membrane potential, recorded in a current-clamp mode. The effects of MβCD were concentration- and time-dependent and reversible. Both development and recovery of the MβCD effects were completed within 1–2 h. Therefore, cholesterol depletion causes significant alterations in the Kv1.3 channel function, whereas T cells possess a potential to reverse these changes.

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Acknowledgements

The authors gratefully acknowledge the technical assistance of P. Oakford (University of Tasmania). This study was supported by grant CONACyT-SEP-2004-CO1-46731 to OD. The costs of academic stay in the University of Tasmania for OD were covered by the General Direction of Academic Exchange (University of Colima).

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Authors and Affiliations

  1. Centro Universitario de Investigaciones Biomédicas (CUIB), Universidad de Colima, Av. 25 de Julio 965, Villa San Sebastian, 28045, Colima, Colima, Mexico

    Igor I. Pottosin, Georgina Valencia-Cruz, Edgar Bonales-Alatorre & Oxana R. Dobrovinskaya

  2. School of Agricultural Science, University of Tasmania, Hobart, Australia

    Sergey N. Shabala

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  1. Igor I. Pottosin
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  2. Georgina Valencia-Cruz
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  3. Edgar Bonales-Alatorre
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  4. Sergey N. Shabala
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  5. Oxana R. Dobrovinskaya
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Correspondence to Oxana R. Dobrovinskaya.

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Pottosin, I.I., Valencia-Cruz, G., Bonales-Alatorre, E. et al. Methyl-β-cyclodextrin reversibly alters the gating of lipid rafts-associated Kv1.3 channels in Jurkat T lymphocytes. Pflugers Arch - Eur J Physiol 454, 235–244 (2007). https://doi.org/10.1007/s00424-007-0208-4

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  • DOI: https://doi.org/10.1007/s00424-007-0208-4

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