A Furosemide-Sensitive Cotransport of Sodium plus Potassium in the Human Red Cell

Wiley, James S.; Cooper, Richard A.

doi:10.1172/jci107613

Cited by 236 publications

(135 citation statements)

References 27 publications

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“…5). These results argue against a K+/K' exchange system described earlier for BALB/c 3T3 cells (21) and fit most easily a Na+K+/Cl-cotransport model as described in other cell types (16)(17)(18)(19)(20).…”

Section: Resultssupporting

confidence: 49%

“…One explanation for the Na' dependence of 86Rb+ uptake (Fig. 3) is a Na+K+/Cl-cotransport system similar to that described in erythrocytes (18,19), L cells (16), Ehrlich ascites cells (17), and epithelial cells (20,22).…”

Section: Resultsmentioning

confidence: 62%

“…The diuretic furosemide is known to inhibit ouabain-insensitive K+ fluxes in several cell types including BALB/c 3T3 cells (16)(17)(18)(19)(20)(21). Fig.…”

Section: Resultsmentioning

confidence: 99%

“…There are at least two possibilities for the nature of the K+ transport system sensitive to PMA and furosemide in these cells: a Na+K+/Cl-cotransport system (16)(17)(18)(19)(20) or a K+/K+ exchange diffusion system (21). To distinguish between these possibilities, four types of experiments were done.…”

Section: Resultsmentioning

confidence: 99%

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Phorbol ester inhibits furosemide-sensitive potassium transport in BALB/c 3T3 preadipose cells.

O’Brien¹,

Krzeminski²

1983

Proc. Natl. Acad. Sci. U.S.A.

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The tumor promoter phorbol 12-myristate 13-acetate (PMA) rapidly decreased the rate of "Rb+ uptake into BALB/c 3T3 preadipose cells. The component of total 86Rb+ influx affected by PMA is insensitive to ouabain but sensitive to the diuretic furosemide. Experiments designed to investigate the characteristics of the K+ transport system sensitive to PMA revealed that: (i) 86Rb+ uptake is highly dependent on external Na+, (ii) "Rb+ uptake is highly dependent on external ClP, (iii) 22Na+ uptake is dependent on external K+, and (iv) a major component of 86Rb+ efflux that is sensitive to PMA and furosemide is not dependent on extracellular KV. These features strongly implicate a Na+K+/Cl-cotransport system as the target of PMA and furosemide in these experiments. PMA caused a net intracellular accumulation of K+ within 15 min in these cells, presumably via its inhibitory effect on furosemide-sensitive K+ transport. Within 30 min after PMA treatment, the mean cell volume was significantly reduced in treated compared to control cells, with a maximum decrease of 21% attained at 4 hr after PMA. The significance of these findings for biologic changes induced by PMA is discussed.In addition to their role in multistage carcinogenesis, the phorbol ester tumor promoters, exemplified by phorbol 12-myristate 13-acetate (PMA), cause a number of biologic and functional changes in diverse cell types in culture (for reviews, see refs. 1 and 2). To date, however, no experimental results to explain any of the actions of tumor promoters in (11), and inhibition of cell differentiation (12). Of particular importance to this study is the observation that PMA is mitogenic in quiescent cultures of this cell line (12), as it is for other 3T3 cell lines (8, 9). In contrast to the earlier work of others (8,9) showing an increased rate of 'Rb+ uptake caused by PMA, we found in these cells that inhibition of 86Rb+ uptake was an early event after PMA treatment. The transport system inhibited by PMA in these cells is insensitive to ouabain and sensitive to the diuretic furosemide, and it apparently catalyzes the cotransport of Na+, K+, and Cl-.MATERIALS AND METHODS Cells. BALB/c 3T3 cells (clone A3IT) were obtained from Leila Diamond (Wistar Institute) and cultured as described (12). The medium used was Eagle Auto-Pow minimal essential medium (Flow Laboratories) supplemented with 10% fetal bovine serum. For flux and ion content experiments, cells were plated from stocks into 35-or 60-mm plastic Petri dishes and used when growing logarithmically or just after reaching confluence. Routinely, cultures were refed with fresh medium every 2-3 days and always on the day prior to experiments.Flux Experiments. The uptake of 'Rb+ was measured in cells maintained in complete culture medium at 37°C in a humidified atmosphere of 95% air/5% CO2. Cells in 35-mm dishes were washed with warm phosphate-buffered saline (unless otherwise stated) and uptake was started by the addition of 2 ml of fresh medium containing 10% fetal bovine serum (K+, 5.4 mM)...

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Section: Resultssupporting

confidence: 49%

Section: Resultsmentioning

confidence: 62%

“…The diuretic furosemide is known to inhibit ouabain-insensitive K+ fluxes in several cell types including BALB/c 3T3 cells (16)(17)(18)(19)(20)(21). Fig.…”

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

Phorbol ester inhibits furosemide-sensitive potassium transport in BALB/c 3T3 preadipose cells.

O’Brien¹,

Krzeminski²

1983

Proc. Natl. Acad. Sci. U.S.A.

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“…1). The ouabain-sensitive component was inhibited by 20 % while the loop diuretic-sensitive and residual fractions (Wiley & Cooper, 1974) [Mg2l]. : mM Fig.…”

Section: Francementioning

confidence: 99%

Communications

1980

The Journal of Physiology

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Chromatolysis in axotomized sympathetic post-ganglionic neurones is accompanied by detachment of their synaptic inputs and loss of post-synaptic membrane densities (Matthews & Nelson, 1975). This suggests a change in neuronal receptor characteristics which might lead to an altered tolerance of the post-synaptic membrane for 'foreign' connexions.To test this hypothesis, the left cervical sympathetic trunk of young male Wistar rats was divided approximately 5 mm caudal to the superior cervical ganglion, under intraperitoneal choral hydrate anaesthesia. The proximal end of the trunk was sutured into the sternomastoid muscle and its distal end was sutured to the proximal end of the divided left hypoglossal nerve. At the same operation, in some animals, the left internal and external carotid nerves were crushed.Between 2 and 14 months later the rats were re-anaesthetized; the hypoglossal nerve and the undisturbed right cervical sympathetic trunk were exposed for stimulation in continuity via platinum electrodes with square wave pulses (10 V, 0-2 msec duration, 5 Hz for 60 sec). The transganglionic effect of stimulation was monitored by the widening of the palpebral fissure, observed in a mirror with a millimetre scale placed beneath the animal's head. Both superior cervical ganglia were then removed and prepared for electron microscopy and counting of synapses.Stimulation of the transposed hypoglossal nerve in eight rats after post-ganglionic nerve crush produced a significantly greater palpebral fissure response than in five rats with intact post-ganglionic nerves (mean widening, 1 6 and 0'36 mm respectively; P < 0 009, Mann-Whitney U-test). The hypoglossal-mediated response was blockable at low dosage by intravenous hexamethonium, by intravenous phentolamine and by nictotine applied directly to the ganglion, but not by intravenous atropine. There was no significant difference between the two groups in palpebral fissure responses to stimulation of the right cervical sympathetic trunk (means, 3-2 and 3*3 mm respectively).The incidence of intraganglionic synapses after hypoglossal nerve transposition was significantly greater (P = 0-003, Mann-Whitney U-test) in seven ganglia after post-ganglionic nerve crush than in seven ganglia with intact post-ganglionic nerves (mean incidences, 1-42 and 0 7 per nucleated neuronal profile respectively; 15P

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A further characterization of the selective K movements observed in human red blood cells following acetylphenylhydrazine exposure

Orringer

1984

American J Hematol

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Following brief exposure to acetylphenylhydrazine, the potassium permeability of the human erythrocyte membrane is selectively augmented. While a similar increase in potassium permeability results from the intracellular accumulation of calcium (the Gardos phenomenon), we have found a number of features that allow these two pathways to be distinguished from one another. The acetylphenylhydrazine pathway does not require calcium for its activation, and can be seen even in the presence of a molar excess of the calcium chelator EGTA. The transmembrane potassium movement via this channel has a specific requirement for the anion chloride, and it can be inhibited by furosemide. The potassium that moves through the Gardos pathway, on the other hand, can be accompanied by any permeant anion, and is inhibitable by quinidine or cetiedil. Thus, acetylphenylhydrazine exposure seems to promote K + Cl cotransport, whereas the Gardos pathway represents a potassium conductive channel. While full demonstration of both these pathways requires harsh in vitro manipulation, the large electrochemical potassium gradient favoring the movement of this cation out from the erythrocyte suggests that even a partial activation of either pathway could cause intracellular dehydration and thus contribute importantly to the pathophysiology of in vivo red cell destruction.

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A Furosemide-Sensitive Cotransport of Sodium plus Potassium in the Human Red Cell

Cited by 236 publications

References 27 publications

Phorbol ester inhibits furosemide-sensitive potassium transport in BALB/c 3T3 preadipose cells.

Phorbol ester inhibits furosemide-sensitive potassium transport in BALB/c 3T3 preadipose cells.

Communications

A further characterization of the selective K movements observed in human red blood cells following acetylphenylhydrazine exposure

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