Abstract
The use of recombinant DNA technology to clone, sequence, and express ion channels and transporters has powered an enormous acceleration in the understanding of structure-function relationships in these important proteins. Given that most ion channels reside in tissues that are largely inaccessible to direct recording techniques and the general paucity of continuous cell lines expressing defined populations of functional molecules, studying native channels is often difficult and impractical. However, the ability to introduce a recombinant complementary DNA (cDNA) selectively into cells normally devoid of highly expressed ion channels or transporters greatly facilitates the ability of scientists to study the function, subunit associations, regulation, and trafficking of these proteins. This approach has also enabled studies designed to investigate the role of ion channel mutations in inherited diseases.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Shih, T. M., Smith, R. D., Toro, L., and Goldin, A. L. (1998) High-level expression and detection of ion channels in Xenopus oocytes. Methods Enzymol. 293, 529–556.
Goldin, A. L. (1992) Maintenance of Xenopus laevis and oocyte injection. Methods Enzymol. 207, 266–279.
Goldin, A. L. and Sumikawa, K. (1992) Preparation of RNA for injection into Xenopus oocytes. Methods Enzymol. 207 279–297.
Stuhmer, W. (1998) Electrophysiologic recordings from Xenopus oocytes. Methods Enzymol. 293, 280–300.
Sherman-Gold, R. (ed.), (1993) The Axon Guide. Axon Instruments, Inc., Foster City, CA, USA.
Barish, M. E. (1983) A transient calcium-dependent chloride current in the immature Xenopus oocyte. J. Physiol. (London) 342, 309–325.
Kowdley, G. C., Ackerman, S. J., John, E. J., Jones, L. R., and Moorman, J. R. (1994) Hyperpolarization-activated chloride currents in Xenopus oocytes. J. Gen. Physiol. 103, 217–230.
Parker, I., and Miledi R. (1988) A calcium-independent chloride current activated by hyperpolarization in Xenopus oocytes. Proc. R. Soc. Lond. B. 233, 191–199.
Landau, E. M., and Blitzer, R. D. (1994) Chloride current assay for phospholipase C in Xenopus oocytes. Methods Enzymol. 238, 140–154.
Tokimasa, T. and North, R. A. (1996) Effects of barium, lanthanum, and gadolinium on endogenous chloride and potassium currents in Xenopus oocytes. J. Physiol. (London) 496, 677–686.
Elsner, H. A., Honck, H. H., Willmann, F., Kreienkamp, H. J., and Iglauer, F. (2000) Poor quality of oocytes from Xenopus laevis used in laboratory experiments: prevention by use of antiseptic surgical technique and antibiotic supplementation. Comp. Med. 50, 206–211.
Eppig, J. J. and Steckman, M. L. (1976) Comparison of exogenous energy sources for in vitro maintenance of follicle cell-free Xenopus laevis oocytes. In Vitro 12, 173–179.
Dumont, J. N. (1972) Oogenesis in Xenopus laevis (Daudin). I. Stages of oocyte development in laboratory maintained animals. J. Morphol. 136, 153–179
Margolskee, R. F., McHendry-Rinde, B., and Horn, R. (1993) Panning transfected cells for electrophysiological studies. BioTechniques 15, 906–911.
Jurman, M. E., Boland, L. M., Liu, Y., and Yellen, G. (1994) Visual identification of individual transfected cells for electrophysiology using antibody-coated beads. BioTechniques 17, 876–881.
Tsien, R. Y. (1998) The green fluorescent protein. Annu. Rev. Biochem. 67, 509–544.
Zhang, G., Gurtu, V., and Kain, S. R. (1996) An enhanced green fluorescent protein allows sensitive detection of gene transfer in mammalian cells. Biochem. Biophys. Res. Commun. 227, 707–711.
Krieg, P. A. and Melton, D. A. (1987) In vitro RNA synthesis with SP6 RNA polymerase. Method Enzymol. 155, 397–415.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc.
About this protocol
Cite this protocol
Tapper, A.R., George, A.L. (2003). Heterologous Expression of Ion Channels. In: Potter, N.T. (eds) Neurogenetics. Methods in Molecular Biology™, vol 217. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-330-5:285
Download citation
DOI: https://doi.org/10.1385/1-59259-330-5:285
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-990-2
Online ISBN: 978-1-59259-330-9
eBook Packages: Springer Protocols