Voltage Clamp Fluorometry: Illuminating the Dynamics of Ion Channels.
Electrophysiology
Fluorescence
Ion channel
RNA microinjection
Two-electrode voltage clamp
Voltage clamp fluorometry
Xenopus laevis
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2024
2024
Historique:
medline:
10
6
2024
pubmed:
10
6
2024
entrez:
10
6
2024
Statut:
ppublish
Résumé
Ion channels comprise one of the largest targets for drug development and treatment and have been a subject of enduring fascination since first discovered in the 1950s. Over the past decades, thousands of publications have explored the cellular biology and molecular physiology of these proteins, and many channel structures have been determined since the late 1990s. Trying to connect the dots between ion channel function and structure, voltage clamp fluorometry (VCF) emerges as a powerful tool because it allows monitoring of the conformational rearrangements underlying the different functional states of the channel. This technique represents an elegant harmonization of molecular biology, electrophysiology, and fluorescence. In the following chapter, we will provide a concise guide to performing VCF on Xenopus laevis oocytes using the two-electrode voltage clamp (TEVC) modality. This is the most widely used configuration on Xenopus oocytes for its relative simplicity and demonstrated success in a number of different ion channels utilizing a variety of attached labels.
Identifiants
pubmed: 38856899
doi: 10.1007/978-1-0716-3818-7_8
doi:
Substances chimiques
Ion Channels
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
119-138Informations de copyright
© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Références
Alexander SPH, Mathie A, Peters JA et al (2021) THE concise guide to pharmacology 2021/22: ion channels. Br J Pharmacol 178
Hodgkin AL, Huxley AF, Katz B (1952) Measurement of current-voltage relations in the membrane of the giant axon of Loligo. J Physiol 116:424–448
doi: 10.1113/jphysiol.1952.sp004716
pubmed: 14946712
pmcid: 1392219
Guan B, Chen X, Zhang H (2013) Two-electrode voltage clamp. In: Gamper N (ed) Ion channels: methods and protocols, Methods in molecular biology, vol 998. Springer, Berlin, pp 79–89
doi: 10.1007/978-1-62703-351-0_6
De Robertis EM, Gurdon JB (2021) A brief history of Xenopus in biology. Cold Spring Harb Protoc 2021:pdb.top107615
Sumikawa K, Houghton M, Emtage JS et al (1981) Active multi-subunit ACh receptor assembled by translation of heterologous mRNA in Xenopus oocytes. Nature 292:862–864
doi: 10.1038/292862a0
pubmed: 7196502
Smart TG, Krishek BJ (1995) Xenopus oocyte microinjection and ion-channel expression. In: Boulton AA, Baker GB, Walz W (eds) Patch-clamp applications and protocols, Neuromethods, vol 26. Humana Press, New Jersey, pp 259–306
doi: 10.1385/0-89603-311-2:259
Mannuzzu LM, Moronne MM, Isacoff EY (1996) Direct physical measure of conformational rearrangement underlying potassium channel gating. Science 271:213–216
doi: 10.1126/science.271.5246.213
pubmed: 8539623
Cha A, Bezanilla F (1997) Characterizing voltage-dependent conformational changes in the K channel with fluorescence. Neuron 19:1127–1140
doi: 10.1016/S0896-6273(00)80403-1
pubmed: 9390525
Sutter Instrument (2020) P-97 flaming/brown™ micropipette puller operation manual, Novato
Nikon (1997) TE-FM Epi-fluorescence attachment instructions. Minato, Tokyo
Warner Instruments OC-725C Oocyte Clamp Amplifier—User’s Manual, Hamden
Molecular Devices (2016) pCLAMP™ 10 data acquisition and analysis software user guide, San José
Westhoff M, Eldstrom J, Murray CI et al (2019) I
doi: 10.1073/pnas.1811623116
pubmed: 30918124
pmcid: 6475427
Sauer M, Hofkens J, Enderlein J (2011) Basic principles of fluorescence spectroscopy. In: Handbook of fluorescence spectroscopy and imaging: from single molecules to ensembles. Wiley, Weinheim, pp 1–30
doi: 10.1002/9783527633500
Lakowicz JR (2006) Principles of fluorescence spectroscopy. Springer, Boston
doi: 10.1007/978-0-387-46312-4
Molecular Devices (2021) Axon guide—electrophysiology and biophysics laboratory techniques, San José
Gandhi CS, Olcese R (2008) The voltage-clamp fluorometry technique. In: Lippiat JD (ed) Potassium channels. Methods in molecular biology, vol 491. Humana Press, Totowa, pp 213–231
doi: 10.1007/978-1-59745-526-8_17
Sanguinetti MC, Curran ME, Zou A et al (1996) Coassembly of KVLQT1 and minK (IsK) proteins to form cardiac IKS potassium channel. Nature 384:80–83
doi: 10.1038/384080a0
pubmed: 8900283
Tammaro P, Shimomura K, Proks P (2008) Xenopus oocytes as a heterologous expression system for studying ion channels with the patch-clamp technique. In: Lippiat JD (ed) Potassium channels. Methods in molecular biology, vol 491. Humana Press, a part of Springer Science + Business Media, Totowa, pp 127–139
doi: 10.1007/978-1-59745-526-8_10
Kalstrup T, Blunck R (2013) Dynamics of internal pore opening in K
doi: 10.1073/pnas.1220398110
pubmed: 23630265
pmcid: 3657800
Warner Instruments Chloriding Ag/AgCl Electrodes Disk, Pellet, or Wire, Hamden