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Effect of guanine nucleotides on the dark voltage of single frog rods

Published online by Cambridge University Press:  02 June 2009

Karl-Friedrich Schmidt ,
Gottfried N. Nöll  and
Christian Baumann
Karl-Friedrich Schmidt
Affiliation:
Physiologisches Institut, Justus-Liebig-Universität Giessen, Federal Republic of Germany
Gottfried N. Nöll
Affiliation:
Physiologisches Institut, Justus-Liebig-Universität Giessen, Federal Republic of Germany
Christian Baumann
Affiliation:
Physiologisches Institut, Justus-Liebig-Universität Giessen, Federal Republic of Germany
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Abstract

Single frog rods consisting of the outer segment and the ellipsoid were investigated by the whole-cell patch-clamp technique. When the recording pipette was filled with a simple intracellular medium containing potassium as the principal cation, a slow increase in dark voltage (hyperpolarization) associated with a decay of the photoresponses was observed. The hyperpolarization started at a dark voltage of −27 ± 8 mV, followed an exponential course, and leveled out at −52 ± 6 mV. The time constant was proportional to the access resistance of the preparations. With a pipette medium containing a 0.5 or 1.0 μM cGMP, the initial dark voltage was shifted to more positive values and the tendency of hyperpolarization was clearly attenuated. Similar results were obtained with 1 mM GTP. The effects of GDP and of ATP were less significant. In experiments with 1 mM GTP plus 1 mM ATP, the dark voltage behaved as in experiments with only GTP. The stabilizing action of GTP was amplified by EGTA so that with 1 mM GTP plus 1 mM free EGTA the dark voltage was stable at a level of −15 mV. It is concluded that the preparations lose intracellular components such as cGMP and GTP by diffusion into the recording pipette and that the losses are prevented or reduced when the pipette medium contains these nucleotides in nearly physiological concentrations. For the internal transmitter cGMP, the results suggest that its free concentration does not exceed 1 μM.

Keywords

FrogPhotoreceptorRetinal rodGuanine nucleotidesPatch clamp
Type
Research Articles
Information
Visual Neuroscience , Volume 2 , Issue 2 , February 1989 , pp. 101 - 108
Copyright
Copyright © Cambridge University Press 1989

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