Switched single-electrode voltage-clamp amplifiers allow precise measurement of gap junction conductance.
Measurement of gap junction conductance (gj) with patch-clamp amplifiers can, due to series resistance problems, be subject to considerable errors when large currents are measured. Formulas developed to correct for these errors unfortunately depend on exact estimates of series resistance, which are not always easy to obtain. Discontinuous single-electrode voltage-clamp amplifiers (DSEVCs) were shown to overcome series resistance problems in single whole cell recording. With the use of two synchronized DSEVCs, the simulated gj in a model circuit can be measured with a maximum error of <5% in all recording situations investigated (series resistance, 5-47 MOmega; membrane resistance, 20-1,000 MOmega; gj, 1-100 nS). At a very low gj of 100 pS, the error sometimes exceeded 5% (maximum of 15%), but the error was always <5% when membrane resistance was >100 MOmega. The precision of the measurements is independent of series resistance, membrane resistance, and gj. Consequently, it is possible to calculate gj directly from Ohm's law, i.e., without using correction formulas. Our results suggest that DSEVCs should be used to measure gj if large currents must be recorded, i.e., if cells are well coupled or if membrane resistance is low.
Müller
A
A
Institute of Pharmacology, University of Cologne, D-50931 Cologne, Germany.
Lauven
M
M
Berkels
R
R
Dhein
S
S
Polder
H R
HR
Klaus
W
W
D000818:Animals; D004553:Electric Conductivity; D004867:Equipment Design; D017629:Gap Junctions / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008564:Membrane Potentials; D008839:Microelectrodes; D008954:Models, Biological; D018408:Patch-Clamp Techniques / Q000295:instrumentation*; D015203:Reproducibility of Results