The Resting Potential of Mouse Leydig Cells: Role of an Electrogenic Na+/K+ Pump

The Resting Potential of Mouse Leydig Cells: Role of an Electrogenic Na+/K+ Pump Resting potentials (Vm) were measured in mouse Leydig cells, using the whole-cell patch-clamp technique. In contrast to conventional microelectrode measurements, where a biphasic potential was observed, we recorded a stable Vm around -32.2 ± 1.2 mV (mean ± SEM, n = 159), at 25°C, and an input resistance larger than 2.7 × 109 W. Although Vm is sensitive to changes in the extracellular concentrations of potassium and chloride, the relationship between Vm and these ions' concentrations cannot be described by either the Goldman-Hodgkin-Katz or the Nernst equation. Perifusing cells with potassium-free solution or 10?3 M ouabain induced a marked depolarization averaging 20.1 ± 3.2 mV (n = 9) and 23.1 ± 2.8 mV, (n = 7), respectively. Removal of potassium or addition of ouabain with the cell voltage-clamped at its Vm, resulted in an inwardly directed current, due to inhibition of the Na+K+ATPase. The pump current increased with temperature with a Q10 coefficient of 2.3 and had an average value of -6.5 ± 0.4 pA (n = 21) at 25°C. Vm also varied strongly with temperature, reaching values as low as -9.2 ± 1.2 mV (n = 22) at 15°C. Taking the pump current at 25°C and a minimum estimate for the membrane input resistance, we can see that the Na+K+ATPase could directly contribute with 17.7 mV to the Vm of Leydig cells, which is a major fraction of the ?32.2 ± 1.2 mV (n = 159) observed. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Journal of Membrane Biology Springer Journals

The Resting Potential of Mouse Leydig Cells: Role of an Electrogenic Na+/K+ Pump

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Publisher
Springer-Verlag
Copyright
Copyright © 2003 by Springer-Verlag New York Inc.
Subject
Life Sciences; Biochemistry, general; Human Physiology
ISSN
0022-2631
eISSN
1432-1424
D.O.I.
10.1007/s00232-002-1048-y
Publisher site
See Article on Publisher Site

Abstract

Resting potentials (Vm) were measured in mouse Leydig cells, using the whole-cell patch-clamp technique. In contrast to conventional microelectrode measurements, where a biphasic potential was observed, we recorded a stable Vm around -32.2 ± 1.2 mV (mean ± SEM, n = 159), at 25°C, and an input resistance larger than 2.7 × 109 W. Although Vm is sensitive to changes in the extracellular concentrations of potassium and chloride, the relationship between Vm and these ions' concentrations cannot be described by either the Goldman-Hodgkin-Katz or the Nernst equation. Perifusing cells with potassium-free solution or 10?3 M ouabain induced a marked depolarization averaging 20.1 ± 3.2 mV (n = 9) and 23.1 ± 2.8 mV, (n = 7), respectively. Removal of potassium or addition of ouabain with the cell voltage-clamped at its Vm, resulted in an inwardly directed current, due to inhibition of the Na+K+ATPase. The pump current increased with temperature with a Q10 coefficient of 2.3 and had an average value of -6.5 ± 0.4 pA (n = 21) at 25°C. Vm also varied strongly with temperature, reaching values as low as -9.2 ± 1.2 mV (n = 22) at 15°C. Taking the pump current at 25°C and a minimum estimate for the membrane input resistance, we can see that the Na+K+ATPase could directly contribute with 17.7 mV to the Vm of Leydig cells, which is a major fraction of the ?32.2 ± 1.2 mV (n = 159) observed.

Journal

The Journal of Membrane BiologySpringer Journals

Published: Jan 1, 2003

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