Validation of the use of the lipophilic thiocyanate anion for the determination of membrane potential in Ehrlich ascites tumor cells |
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Authors: | Thomas C Smith Susan C Robinson |
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Institution: | (1) Department of Physiology, University of Texas Health Science Center, 78284 San Antonio, Texas |
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Abstract: | Summary The utility of the lipophilic anion thiocyanate (SCN+) as a probe for the indirect estimation of the cell membrane potential (V
m
) in Ehrlich ascites tumor cells has been evaluated by comparison to direct electrophysiological measurements. SCN accumulation is consisten with first-order uptake into a single kinetically-identifiable cellular compartement, achieving steadystate distribution in 20–30 min at 22°C. The steady-state distribution ratio (SCN–]
c
/SCN–]
e
) in physiological saline is 0.44±0.02. Treatment of the cells with proparanolol (0.13 mM), an activator of Ca2+ dependent K+ channels, reduces the steady-state distribution ratio to 0.19±0.02. Conversely, treatmetn with BACl2 (10 mM), an antagonist of the pathway, increases the SCN– distribution ratio to 0.62±0.01. The equilibrium potentials (V
SCN
) calculated under these conditions are virtually identical to direct electrophysiological measurements of theV
m
made under the same conditions. The effect of varing extracellular K+](K+]
e
) in the presence of constant Na+]
e
=100 mM has also been tested. In control cells, elevation of K+]
e
from 6 to 60 mM reducesV
SCN
from –20.6±1.0 to –13.2±1.2 mV. Again, microelectrode measurements give excellent quantitative agreement. Propranolol increases the sensitivity of the cells to varying K+]
e
, so that a 10-fold elevation reducesV
SCN
by approximately 31 mV. BaCl2 greatly reduces this reponse: a 10-fold elevation in K+]
e
yielding only a 4-mV rediction inV
SCN
. It is concluded that the membrane potential of Ehrlich cells can be estimated accurately from SCN– distribution measurements. |
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Keywords: | Ehrlich ascites tumor cell membrane potential thiocyanate K+ channels barium propranlol lipophilic anions |
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