Novel voltage clamp to record small, fast currents from ion channels expressed in Xenopus oocytes.
The present report describes a novel technique for voltage-clamping amphibian oocytes in which part of the membrane is isolated by a vaseline gap and the cytoplasmic fluid is exchanged by cutting or permeabilizing the remaining membrane. The main features of this open-oocyte, vaseline-gap voltage clamp are: (a) low current noise (1 nA at 3 kHz), (b) control of the ionic composition of both the internal and external media, (c) fast time resolution (20-100 microseconds time constant of decay of the capacity transient) and (d) stable recordings for several hours. These features allow reliable measurements of tail or gating currents and the new method is especially suitable when either of these currents must be measured to test the effects of mutations introduced into the cDNAs of cloned ion channels.
Taglialatela
M
M
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX 77030.
Toro
L
L
Stefani
E
E
D000818:Animals; D001921:Brain / Q000502:physiology; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods; D015723:Gene Library; D066298:In Vitro Techniques; D015640:Ion Channel Gating; D008564:Membrane Potentials; D009865:Oocytes / Q000502:physiology*; D015221:Potassium Channels / Q000235:genetics / Q000502:physiology*; D051381:Rats; D015222:Sodium Channels / Q000235:genetics / Q000502:physiology*; D014981:Xenopus
D015221:Potassium Channels; D015222:Sodium Channels