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Heterologous expression of the human potassium channel Kv2.1 in clonal mammalian cells by direct cytoplasmic microinjection of cRNA

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  • Molecular and Cellular Physiology
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Summary

The cloned human delayed rectifying K+ channel Kv2.1 (drk1) was expressed in clonal mouse fibroblasts (L-cells) and rat basophilic leukemia cells (RBL-1) by direct cytoplasmic microinjection of complementary RNA (cRNA). Within six hours, cells microinjected with Kv2.1 cRNA expressed a large sustained outward current as determined from whole-cell patch-clamp recordings. Nearly 100% of cells injected with cRNA expressed outward current. Current density was 30–70 pA/pF when measured at a potential of +50 mV. Steady-state activation and inactivation parameters for Kv2.1 were similar when expressed in either L-cells or RBL-1 cells. These results are the first to demonstrate that functional ion channel proteins can be expressed in mammalian clonal cell lines by direct cytoplasmic microinjection of cRNA.

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References

  • Frech GC, VanDongen AMJ, Shuster G, Brown AM, Joho RH (1989) A novel potassium channel with delayed rectifier properties isolated from rat brain by expression cloning. Nature 340: 642–645.

    Google Scholar 

  • Hosoi S, Slayman, CL (1991) Membrane voltage, resistance, and channel switching in isolated mouse fibroblasts (L-cells): A patch-electrode analysis. J Physiol 367: 267–290.

    Google Scholar 

  • Joho RH, Moorman, JR, VanDongen AMJ, Kirsch GE, Silberberg H, Schuster G and Brown AM (1990) Toxin and kinetic profile of rat brain type III sodium channel expressed in Xenopus oocytes. Mol Brain Research 7:105–113.

    Google Scholar 

  • Lindau M, Fernandez JM (1986) Patch-clamp study of histamine secreting cells. J Gen Physiol 88: 349–368.

    Google Scholar 

  • Sumikawa K, Houghton M, Emtage JS, Richards BM, Barnard EA (1981) Active multi-unit ACh receptor assembled by translation of heterologous mRNA in Xenopus oocytes. Nature 292: 862–864.

    Google Scholar 

  • Taglialatela M, VanDongen, AMJ, Drewe, JA, Joho, RH, Brown, AM, Kirsch, GE (1992) Patterns of internal and external letraethylammonium block in four homologous K+ channels. Mol Pharmacol 40: 229–307.

    Google Scholar 

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Author notes

  1. Fernando Soler

    Present address: Departamento de Bioquímica y Biología Molecular A, Universidad de Murcia, Murcia, Spain

  2. Roger D. Zühlke & Rolf H. Joho

    Present address: Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center at Dallas, 75235, Dallas, Texas, USA

Authors and Affiliations

  1. Department of Pharmacology and Toxicology, Medical College of Georgia, 30912, Augusta, GA, USA

    Stephen R. Ikeda & Deborah L. Lewis

  2. Department of Molecular Physiology and Biophysics, Baylor College of Medicine, 77030, Houston, TX, USA

    Fernando Soler, Roger D. Zühlke & Rolf H. Joho

Authors
  1. Stephen R. Ikeda
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  2. Fernando Soler
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  3. Roger D. Zühlke
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  4. Rolf H. Joho
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  5. Deborah L. Lewis
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Ikeda, S.R., Soler, F., Zühlke, R.D. et al. Heterologous expression of the human potassium channel Kv2.1 in clonal mammalian cells by direct cytoplasmic microinjection of cRNA. Pflügers Arch 422, 201–203 (1992). https://doi.org/10.1007/BF00370422

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  • DOI: https://doi.org/10.1007/BF00370422

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