Electrically silent cotransport of Na+, K+ and Cl− in ehrlich cells

Geck, Peter; Pietrzyk, C.; Burckhardt, Birgitta-Christina; Pfeifferl, B.; Heinz, Erich

doi:10.1016/0005-2736(80)90446-0

Cited by 476 publications

(211 citation statements)

References 21 publications

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“…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: 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%

See 1 more Smart Citation

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 ionic requirements for cell shrinkage by 8-Br-cGMP match those for Na+/K+/2CI -cotransport (Geck et al, 1980;O'Grady et al, 1987). Replacing Na + or C1-with ions that are not transported should preclude further inhibition of transport by 8-Br-cGMP.…”

Section: Does Cgmp Reduce Cell Volume By Inhibiting Na+/k+/2cl -Cotramentioning

confidence: 89%

“…Na+/K+/2CI-cotransport requires extracellular Na + and CI-, and removal of these ions from the bathing solution blocks the cotransport process (Geck, Pietrzyk, Burckhardt, Pfeiffer, and Heinz, 1980;O'Grady et al, 1987). If 8-Br-cGMP shrinks cells by blocking Na+/K+/2CI -cotransport, it should have no effect on cells that are bathed in either Na +-or C1--free media since Na+/K+/2CI-cotransport would have already been inhibited.…”

Section: Cgmp Blocks Na+ /K+ /2cl -Cotransportmentioning

confidence: 99%

Modulation of rabbit ventricular cell volume and Na+/K+/2Cl- cotransport by cGMP and atrial natriuretic factor.

Clemo

Feher

Baumgarten

1992

The Journal of General Physiology

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Previously we showed that atrial natriuretic factor (ANF) decreases cardiac cell volume by inhibiting ion uptake by Na+/K+/2CI-cotransport. Digital video microscopy was used to study the role of guanosine 3',5'-monophosphate (cGMP) in this process in rabbit ventricular myocytes. Each cell served as its own control, and relative cell volumes (volumetest/volumeco,trol) were determined. Exposure to 10 IxM 8-bromo-cGMP (8-Br-cGMP) reversibly decreased cell volume to 0.892 -0.007; the EDs0 was 0.77 +-0.33 tzM. Activating guanylate cyclase with 100 IxM sodium nitroprusside also decreased cell volume to 0.889 ---0.009. In contrast, 8-bromo-adenosine 3',5'-monophosphate (8-Br-AMP; 0.01-100 tzM) neither altered cell volume directly nor modified the response to 8-Br-cGMP. The idea that cGMP decreases cell volume by inhibiting Na+/K+/2C1-cotransport was tested by blocking the cotransporter with 10 I~M bumetanide (BUM) and removing the transported ions. After BUM treatment, 10 IxM 8-Br-cGMP failed to decrease cell volume. Replacement of Na ÷ with N-methyl-D-glucamine or C1-with methanesulfonate also prevented 8-Br-cGMP from shrinking cells. The data suggest that 8-Br-cGMP, like ANF, decreases ventricular cell volume by inhibiting Na+/K+/2C1-cotransport. Evidence that ANF modulates cell volume via cGMP was also obtained. Pretreatment with 10 IzM 8-Br-cGMP prevented the effect of 1 IzM ANF on cell volume, and ANF suppressed 8-Br-cGMP-induced cell shrinkage. Inhibiting guanylate cyclase with the quinolinedione LY83583 (10 tzM) diminished ANF-induced cell shrinkage, and inhibiting cGMP-specific phosphodiesterase with M&B22948 (Zaprihast; 100 IzM) amplified the volume decrease caused by a low dose of ANF (0.01 I~M) approximately fivefold. In contrast, neither 100 I~M 8-Br-cAMP nor 50 tzM forskolin affected the response to ANF. The effects of ANF, LY83583, and M&B29948 on cGMP levels in isolated ventricular myocytes were confirmed by 125I-cGMP radioimmunoassay. These data argue that ANF shrinks cardiac cells by increasing intracellular cGMP, thereby inhibiting Na+/K+/2C1-cotransport. Basal cGMP levels also appear to modulate cell volume.

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Potassium homeostasis and glial energy metabolism

Amédée

Robert²,

Coles

1997

Glia

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Since capillaries appear not to contribute significantly to rapid removal of K+ from brain tissue, the K+ released into extracellular clefts by neurons at the onset of electrical activity is presumably removed either by redistribution in the clefts or by uptake into cells. What appear to be the three major processes require no energy from the glial cells. These are diffusion through the extracellular clefts, spatial buffering by glial cells, and net uptake of K+ into glial cells through glial K+ channels associated with uptake of Cl−through an independent Cl‐ conductance. There is a relatively slow uptake by the Na+/K+‐ATPase, which directly consumes ATP. In addition, some glial cells take up K+ on the Na+/K+/2Cl‐ cotransporter, which leads indirectly to energy consumption when the Na+ is subsequently pumped out. Currently available data suggest that the glial energy metabolism devoted to K+ homeostasis is less than a tenth of the total tissue energy metabolism, even under conditions of pathologically high extracellular [K+]. Hence, in situ, it is possible that glial cells could function with much less ATP than neurons do. All the various routes of muffling of changes in extracellular [K+] can be modulated, directly or indirectly, by transmitters liberated by neurons. A consequence of this could be regulation of the entry of Na+ into glial cells such that the Na+/K+‐ATPase is activated. The degree of activation might be adjusted so that the resulting activation of the glial glycolytic pathway is appropriate to the provision of the quantity of metabolic substrates required by the neurons. GLIA 21:46–55, 1997. © 1997 Wiley‐Liss, Inc.

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Electrically silent cotransport of Na+, K+ and Cl− in ehrlich cells

Cited by 476 publications

References 21 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.

Modulation of rabbit ventricular cell volume and Na+/K+/2Cl- cotransport by cGMP and atrial natriuretic factor.

Potassium homeostasis and glial energy metabolism

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