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R M Spanswick

Publications and source records attributed to R M Spanswick.

43 records · Page 3Linked to original sources

Correlation of Adenosine Triphosphate Levels in Chara corallina with the Activity of the Electrogenic Pump.

The effect of a number of inhibitors on the ATP level in single cells of Chara corallina has been measured using the luciferin-luciferase assay. The uncouplers of phosphorylation, carbonyl cyanide m-chlorophenyl hydrazone and 2,4-dinitrophenol, and the ATPase inhibitors, dicyclohexyl-carbodimide and diethylstilbestrol, all caused a marked reduction of the ATP level. These inhibitors also produced a large increase in the membrane resistance and a depolarization of the membrane potential to the diffusion potential. This is consistent with the plasmalemma containing an ATP-dependent electrogenic pump that provides the primary conductance through the membrane.Ethyl-3-(3-dimethylaminopropyl)carbodiimide, which depolarizes the membrane potential, has no effect on the ATP level and does not increase the membrane resistance. This inhibitor apparently does not enter the cell but may act by affecting the permeability of the membrane. Neither darkness nor 3-(3,4-dichlorophenyl)-1,1-dimethylurea lowers the ATP level and, while neither has much effect on the membrane potential, both cause a similar increase in resistance in comparison with the control in the light. The weak acid, 5,5-dimethyloxazolidine-2,4-dione, and the weak base, NH(3), do not affect the ATP level significantly but have effects on the electrogenic pump that are consistent with their postulated effects on the cytoplasmic pH, if H(+) is the substrate for the pump.

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Activity of the Electrogenic Pump in Chara corallina as Inferred from Measurements of the Membrane Potential, Conductance, and Potassium Permeability.

The effects of various inhibitors on the membrane potential, resistance, and K(+) permeability of Chara corallina were measured, providing evidence that there is an electrogenic pump in the membrane. It was found that: (a) 5.0 mum carbonyl cyanide m-chlorophenyl hydrazone depolarizes the membrane potential and increases the membrane resistance. This inhibition is faster in the dark than in the light but the extent of inhibition is the same in both cases. (b) Fifty mum dicyclohexylcarbodiimide increases the resistance and the K(+) permeability and depolarizes the membrane to a diffusion potential mainly controlled by K(+). (c) Forty mum diethylstilbestrol and 0.1 mm 2,4-dinitrophenol increase the resistance and depolarize the potential to a value given by the Goldman diffusion equation. (d) Both 3-(3,4-dichlorophenyl)-1,1-dimethylurea and darkness (at pH 6) cause the membrane resistance to increase but neither has a large effect on the potential. 3-(3,4-dichlorophenyl)-1,1-Dimethylurea increases K(+) permeability while darkness decreases it.In all cases, the increase in resistance is interpreted as an inhibition of conductance through the electrogenic pump. As a consequence of this inhibition, the electrogenic component of the membrane potential is reduced, depolarizing the membrane. The electrogenic pump may be an H(+)-ATPase in the plasmalemma.5,5-Dimethyloxazolidine-2,4-dione at 5.0 mm decreases the membrane resistance, by lowering the internal pH providing more substrate for the pump. La(3+) decreased cation permeability and depolarized the membrane but, since it had little effect on the membrane resistance, it probably does not affect the electrogenic pump.

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Measurement of the Cytoplasmic pH in Nitella translucens: Comparison of Values Obtained by Microelectrode and Weak Acid Methods.

A comparison has been made between the use of two types of pH microelectrode and the weak acid method for determining the cytoplasmic pH of Nitella translucens at an external pH of 6. There was good agreement between the value obtained with glass pH microelectrodes (7.54 +/- 0.15 se) and that obtained using the weak acid 5,5-dimethyloxazolidine-2,4-dione (7.42 +/- 0.07 se). Plastic-insulated antimony microelectrodes gave a significantly lower value (6.74 +/- 0.15 se) possibly due to disruption of the insulation by the cell wall. The addition of 1 mM NaN(3) rapidly reduced the pH recorded by the glass pH microelectrodes to about 5.3. A smaller change was observed using the weak acid method. The relevance of this observation to recent work on indoleacetic acid transport is discussed.

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