Abstract
The Xenopus laevis oocyte offers one of the most convenient expression systems for assaying the actions of candidate ligands on cloned ionotropic neurotransmitter receptors (also known as ligand-gated ion channels [LGICs]). Their large size makes injection of complementary ribonucleic acid or complementary deoxyribonucleic acid and electro-physiological recording very easy. Furthermore, Xenopus oocytes translate messages very efficiently, resulting in the detection of large-amplitude ligand-induced currents from expressed, recombinant LGICs. Compared to other electrophysiological techniques, recording from oocytes is not difficult and requires only a basic electrophysiological recording setup. Oocytes can be used for two-electrode voltage clamp, as well as cell-attached patch and inside- or outside-out patch clamp recordings. A variety of protocols allows the experimenter to determine the actions of ligands on cloned receptors and parameters, such as their affinity, efficacy, rates of association and desensitization, and reversibility, to be estimated. Here, we present protocols for using Xenopus oocytes in assaying candidate ligands acting against cloned targets of drugs and pesticides.
Key Words
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Methfessel, C, Witzemann, V., Takahashi, T., Mishina, M., Numa, S., and Sakmann, B. (1986) Patch clamp measurements on Xenopus laevis oocytes: currents through endogenous channels and implanted acetylcholine receptor and sodium channels. Pflugers Arch. 407, 577–588.
Dempster, J. (1993) Computer Analysis of Electrophysiological Signals, Academic Press, London.
Arena, J. P., Liu, K. K., Paress, P. S., Schaeffer, J. M., and Cully, D. F. (1992) Expression of a glutamate-activated chloride current in Xenopus oocytes injected with Caenorhabditis elegans RNA: evidence for modulation by avermectin. Brain Res. Mol. Brain Res. 15, 339–348.
Njue, A. I., Hayashi, J., Kinne, L., Feng, X. P., and Prichard, R. K. (2004) Mutations in the extracellular domains of glutamate-gated chloride channel α3 and β subunits from ivermectin-resistant Cooperia oncophora affect agonist sensitivity. J. Neurochem. 89, 1137–1147.
Hosie, A. M., Baylis, H. A., Buckingham, S. D., and Sattelle, D. B. (1995) Actions of the insecticide fipronil, on dieldrin-sensitive and-resistant GABA receptors of Drosophila melanogaster. Br. J. Pharmacol. 115, 909–912.
Buckingham, S. D., Hosie, A. M., Roush, R. L., and Sattelle, D. B. (1994) Actions of agonists and convulsant antagonists on a Drosophila melanogaster GABA receptor (RDL) homo-oligomer expressed in Xenopus oocytes. Neurosci. Lett. 181, 137–140.
Bali, M. and Akabas, M. H. (2004) Defining the propofol binding site location on the GABAA receptor. Mol. Pharmacol. 65, 68–76.
Murasaki, O., Kaibara, M., Nagase, Y., Mitarai, S., Doi, Y., Sumikawa, K., and Taniyama, K. (2003) Site of action of the general anesthetic propofol in muscarinic Ml receptor-mediated signal transduction. J. Pharmacol. Exp. Ther. 307, 995–1000.
Do, S. H., Ham, B. M., and Zuo, Z. (2003) Effects of propofol on the activity of rat glutamate transporter type 3 expressed in Xenopus oocytes: the role of protein kinase C. Neurosci. Lett. 343. 113–116.
Davies, M., Newell. J. G.. Derry, M., Martin, I. L., and Dunn, S. M. Characterization of the interaction of zopiclone with γ-aminobutyric acid type A receptors. Mol. Pharmacol. 58, 756–762.
Okamoto, T., Minami, K., Uezono, Y., et al. (2003) The inhibitory effects of ketamine and pentobarbital on substance P receptors expressed in Xenopus oocytes. Anesth. Analg. 97, 104–110.
Levandoski, M. M., Piket, B., and Chang, J. (2003) The anthelmintic levamisole is an allosteric modulator of human neuronal nicotinic acetylcholine receptors. Eur. J. Pharmacol. 471, 9–20.
Raymond Delpech, V., Ihara, M., Coddou, C, Matsuda, K., and Sattelle, D. B. (2003) Action of nereistoxin on recombinant neuronal nicotinic acetylcholine receptors expressed in Xenopus laevis oocytes. Invert. Neurosci. 5. 29–35.
Nishiwaki, H., Nakagawa, Y., Kuwamura, M., et al. (2003) Correlations of the electro-physiological activity of neonicotinoids with their binding and insecticidal activities. Pest. Manag. Sci. 59, 1023–1030.
Ihara, M., Matsuda, K., Otake, M., et al. (2003) Diverse actions of neonicotinoids on chicken alpha7, alpha4beta2 and Drosophila-chicken SADbeta2 and ALSbeta2 hybrid nicotinic acetylcholine receptors expressed in Xenopus laevis oocytes. Neuropharmacology 45, 133–144.
Ogata, J., Minami, K., Uezono, Y., et al. (2004) The inhibitory effects of tramadol on 5-hydroxytryptamine type 2C receptors expressed in Xenopus oocytes. Anesth. Analg. 98, 1401–1406.
Wagner, L. E., Gingrich, K. J., Kulli, J. C, and Yang, J. (2001) Ketamine blockade of voltage-gated sodium channels: evidence for a shared receptor site with local anesthetics. Anesthesiology 95, 1406–1413.
Cheffings, C. M. and Colquhoun, D. (2000) Single channel analysis of a novel NMDA channel from Xenopus oocytes expressing recombinant NR1 A, NR2A and NR2D subunits. J. Physiol. 526Pt. 3, 481–491.
Hosie, A. M., Buckingham, S. D., Presnail, J. K., and Sattelle, D. B. (2001) Alternative splicing of a Drosophila GABA receptor subunit gene identifies determinants of agonist potency. Neuroscience 102, 709–714.
Shirai, Y., Hosie, A. M., Buckingham, S. D., Holyoke, C. W., Jr., Baylis, H. A., and Sattelle, D. B. (1995) Actions of picrotoxinin analogs on an expressed, homo-oligomeric GABA receptor of Drosophila melanogaster. Neurosci. Lett. 189, 1–4.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Buckingham, S.D., Pym, L., Sattelle, D.B. (2006). Oocytes as an Expression System for Studying Receptor/Channel Targets of Drugs and Pesticides. In: Liu, X.J. (eds) Xenopus Protocols. Methods in Molecular Biology™, vol 322. Humana Press. https://doi.org/10.1007/978-1-59745-000-3_23
Download citation
DOI: https://doi.org/10.1007/978-1-59745-000-3_23
Publisher Name: Humana Press
Print ISBN: 978-1-58829-362-6
Online ISBN: 978-1-59745-000-3
eBook Packages: Springer Protocols