Medizinische Universität Graz - Research portal
Selected Publication:
Schreibmayer, W; Lester, HA; Dascal, N.
Voltage clamping of Xenopus laevis oocytes utilizing agarose-cushion electrodes.
Pflugers Arch. 1994; 426(5):453-458
Doi: 10.1007/BF00388310
Web of Science
PubMed
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- Leading authors Med Uni Graz
- Schreibmayer Wolfgang
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- Abstract:
- Two-electrode voltage clamping of expressed ion channels in intact oocytes of the South African clawed frog Xenopus laevis has been refined to allow stable, low-resistance electrical access to the cytosol (50-800 k omega). Glass microelectrodes were filled with a cushion of 1% agarose at their tips to prevent KCl leakage (agarose-cushion electrodes). Insertion of these electrodes into X. laevis oocytes yielded stable preparations for periods of more than 1 h with a stable input resistance of 1-4 M omega. Furthermore, a simple modification of the voltage-clamp circuit (charging compensator) is described that increases the flexibility of arrangements for differential recording of the membrane potential in order to subtract voltage drops across a series resistance. The result is a considerable increase in the practically attainable speed of the voltage clamp with the conventional two-electrode arrangement. The performance of the charging compensator was tested on an equivalent circuit that simulates the oocyte and electrodes. In addition, the combination of agarose-cushion electrodes and the charging compensator was tested on oocytes expressing Shaker H4 currents. The fidelity of the voltage-clamp circuit was also verified by measuring the membrane potential with additional independent microelectrodes connected to a differential amplifier, independent of the two-electrode voltage clamp system. The system described here will be useful for ion channel studies in X. laevis oocytes requiring long-term recordings and/or measurements of large, fast ion currents.
- Find related publications in this database (using NLM MeSH Indexing)
- Animals -
- Cytosol - physiology
- Electric Stimulation - physiology
- Female - physiology
- Ion Channels - physiology
- Microelectrodes - physiology
- Models, Biological - physiology
- Oocytes - physiology
- Sepharose - physiology
- Xenopus laevis - physiology
- Find related publications in this database (Keywords)
- Agarose-Cushion Electrodes
- Charging Compensator
- 2-Electrode Voltage Clamp
- Xenopus Laevis Oocytes
- Ion Channel Modulation
- Heterologous Expression