Expression and study of ligand‐gated ion channels in Xenopus laevis oocytes
Kapur, A.; Derry, J.M.C.; Hansen, R.S. (2007). Expression and study of ligand‐gated ion channels in Xenopus laevis oocytes, in: Lajtha, A. et al. Handbook of neurochemistry and molecular neurobiology: Practical neurochemistry methods. pp. 323-340. https://dx.doi.org/10.1007/978-0-387-30401-4_14
In: Lajtha, A. et al. (Ed.) (2007). Handbook of neurochemistry and molecular neurobiology: Practical neurochemistry methods. Springer Science and Business Media: Boston. ISBN 9780387303598. , more
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| Authors | | Top |
- Kapur, A.
- Derry, J.M.C.
- Hansen, R.S.
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| Abstract |
The study of neurotransmitter ion channels has been greatly facilitated by use of the Xenopus laevis oocyte expression system. The reliable expression of exogenous receptors in the Xenopus oocyte, including those belonging to the ligand‐gated ion channel family, has provided a powerful means for investigating structure‐function relationships in these receptors using a variety of methods such as two‐electrode voltage clamp electrophysiology, patch‐clamp studies, and radioligand binding assays. The following chapter outlines morphology of Xenopus laevis frogs and oocytes, and reviews the advantages and limitations of using Xenopus oocytes to study exogenously expressed proteins. Techniques for oocyte isolation and preparation, in vitro preparation and injection of RNA transcripts, and subsequent functional analysis of expressed ligand‐gated ion channel receptors which include the nAChR, the GABAA receptor, the 5‐HT3 and glycine receptors, are discussed in detail. The use of the two‐electrode voltage clamp setup to characterize whole‐cell receptor currents, and the types of concentration‐effect experiments that can be performed using this arrangement, are reviewed. Additional uses of Xenopus oocytes in characterizing the structure‐activity relationship of these receptors are also explored. |
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