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J Neurophysiol (January 9, 2008). doi:10.1152/jn.01232.2007
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Submitted on November 8, 2007
Accepted on January 8, 2008

Space clamp problems when voltage clamping neurons expressing voltage-gated conductances

Dan Bar-Yehuda1 and Alon Korngreen1*

1 Faculty of Life Sciences, Bar-Ilan Univ., Israel, Ramat-gan, Israel

* To whom correspondence should be addressed. E-mail: korngra{at}mail.biu.ac.il.

The voltage-clamp technique is applicable only to spherical cells. In non-spherical cells, such as neurons, the membrane potential is not clamped distal to the voltage-clamp electrode. This means that the current recorded by the voltage-clamp electrode is the sum of the local current and of axial currents from locations experiencing different membrane potentials. Furthermore, voltage-gated currents recorded from a non-spherical cell are, by definition, severely distorted due to the lack of space-clamp. Justifications for voltage-clamping in non-spherical cells are, firstly, that the lack of space-clamp is not severe in neurons with short dendrites. Secondly, passive cable theory may be invoked to justify application of voltage-clamp to branching neurons, suggesting that the potential decay is sufficiently shallow to allow spatial clamping of the neuron. Here, using numerical simulations, we show that the distortions of voltage-gated K+ and Ca2+ currents are substantial even in neurons with short dendrites. The simulations also demonstrate that passive cable theory cannot be used to justify voltage-clamping of neurons, due to significant shunting to the reversal potential of the voltage-gated conductance during channel activation. Some of the predictions made by the simulations were verified using somatic and dendritic voltage-clamp experiments in rat somatosensory cortex. Our results demonstrate that voltage-gated K+ and Ca2+ currents recorded from branching neurons are almost always severely distorted.







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