Evidence for the genetic control of the sodium pump density in HeLa cells

Boardman, L. J.; Huett, Marian; Lamb, J. F.; Newton, James P.; J, Polson

doi:10.1113/jphysiol.1974.sp010684

Cited by 105 publications

(61 citation statements)

References 31 publications

(26 reference statements)

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“…These findings are qualitatively similar to those in HeLa cells (Boardman et al, 1974(Boardman et al, , 1975Pollack et al, 1981). However, the concentrations of lithium which produce this effect are lower in the lymphocyte (1 mmol 1-1) than in the HeLa cell (20-50 mmol 1-1) (Boardman et al, 1975;Aiton, 1976).…”

Section: Discussionsupporting

confidence: 80%

The effects of serum, lithium, ethacrynic acid, and a low external concentration of potassium on specific [3H]‐ouabain binding to human lymphocytes after incubation for 3 days.

Rapeport¹,

Aronson²,

Grahame‐Smith³

et al. 1986

Brit J Clinical Pharma

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We have quantified specific [3H]‐ouabain binding sites in normal human lymphocytes, and have measured the changes in the numbers of those sites which occur in response to various stimuli. We have confirmed previous findings that incubation for 72 h in the presence of fetal calf serum causes an increase in [3H]‐ouabain binding, and that this does not occur if the cells are incubated in fetal calf serum which has first been dialysed. During incubation of the lymphocytes for 3 days in the presence of dialysed fetal calf serum each of the following stimuli caused an increase in specific [3H]‐ouabain binding: addition of ethacrynic acid (1 mumol l‐1), addition of lithium (1 mmol l‐1), and reduction of the external potassium concentration (to 0.75 mmol l‐1). By analogy with the similar results in HeLa cells reported by others, we suggest that the increase in [3H]‐ouabain binding may, in the case of ethacrynic acid and the reduction of the external potassium concentration, be initiated by an increase in the intracellular sodium concentration. The mechanisms whereby fetal calf serum and lithium cause an increase in [3H]‐ouabain binding are not clear.

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Section: Discussionsupporting

confidence: 80%

The effects of serum, lithium, ethacrynic acid, and a low external concentration of potassium on specific [3H]‐ouabain binding to human lymphocytes after incubation for 3 days.

Rapeport¹,

Aronson²,

Grahame‐Smith³

et al. 1986

Brit J Clinical Pharma

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“…There is evidence in a number of cell systems that the level of expression (i.e., number per unit area) of Na pumps in cell membranes is under the control of Na i , such that an increase in Na i leads to an increase in their expression (34,35). This finding could represent an adaptative response to an increase in Na i by creating the circumstance to lower Na i by increased Na export.…”

Section: Discussionmentioning

confidence: 87%

Na pump isoforms in human erythroid progenitor cells and mature erythrocytes

Hoffman

Wickrema

Potapova

et al. 2002

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

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This study is aimed at identifying the Na pump isoform composition of human erythroid precursor cells and mature human erythrocytes. We used purified and synchronously growing human erythroid progenitor cells cultured for 7-14 days. RNA was extracted from the progenitor cells on different days and analyzed by RT-PCR. The results showed that only the ␣1, ␣3, ␤2, and ␤3 subunit isoforms and the ␥ modulator were present. Northern analysis of the erythroid progenitor cells again showed that ␤2 but not ␤1 or ␣2 isoforms were present. The erythroid cells display a unique ␤ subunit expression profile (called ␤-profiling) in that they contain the message for the ␤2 isoform but not ␤1, whereas leukocytes and platelets are known to have the message for the ␤1 but not for the ␤2 isoform. This finding is taken to indicate that our preparations are essentially purely erythroid and free from white cell contamination. Western analysis of these cultured progenitor cells confirmed the presence of ␣1, ␣3, (no ␣2), ␤2, ␤3, and ␥ together now with clear evidence that ␤1 protein was also present at all stages. Western analysis of the Na pump from mature human erythrocyte ghosts, purified by ouabain column chromatography, has also shown that ␣1, ␣3, ␤1, ␤2, ␤3, and ␥ are present. Thus, the Na pump isoform composition of human erythroid precursor cells and mature erythrocytes contains the ␣1 and ␣3 isoforms of the ␣ subunit, the ␤1, ␤2, and ␤3 isoforms of the ␤ subunit, and the ␥ modulator.

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“…This concentration of cycloheximide is sufficient to inhibit protein synthesis by > 95% (Boardman et al, 1974). Because the half time of dissociation of ouabain from the pump site is very slow (approximately 20 h, Boardman et al, 1972) and independent of protein synthesis, the cycloheximide-sensitive component of recovery indicates the appearance of newly synthesized sodium pump sites.…”

Section: Recoverymentioning

confidence: 99%

“…The fraction of the 86Rb influx catalysed by the sodium pump (the ouabain sensitive 86Rb influx) was taken as the difference between the 86Rb influxes measured in the presence and absence of 10-3M ouabain. The techniques used in this study have been described in detail elsewhere Boardman, et al, 1972;Boardman, Huett, Lamb, Newton & Polson, 1974;Aiton & Lamb, 1980) and are summarised below. All experiments were carried out at 37°C.…”

Section: Cell Culturementioning

confidence: 99%

DOWN‐REGULATION OF THE SODIUM PUMP FOLLOWING CHRONIC EXPOSURE OF HeLa CELLS AND CHICK EMBRYO HEART CELLS TO OUABAIN

Aiton

Lamb

Ogden

1981

British J Pharmacology

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HeLa cells and primary cultures of embryonic chick heart cells were grown in medium containing low concentrations of ouabain for 24 h. Compared with normal cells, cells grown in ouabain have fewer free sodium pump sites, an increased intracellular sodium concentration and a decreased intracellular potassium concentration. The cells are able to maintain their intracellular ion contents because the remaining pump sites have an increased turnover rate. When cells that have been chronically exposed to ouabain are returned to normal growth medium, the sodium pump site numbers increase; the recovery process begins within 6 to 8 h and is complete within 24 h. Recovery of pump site numbers is primarily dependent upon de novo protein synthesis since the protein synthesis inhibitor, cycloheximide, prevents recovery.

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Evidence for the genetic control of the sodium pump density in HeLa cells

Abstract: SUMMARY1. HeLa cells were grown in normal and altered growth solutions; the ion contents, volumes, K sensitive ouabain binding, the Na-K-ATPase and the Na and K transport measured.2.

Cited by 105 publications

References 31 publications

The effects of serum, lithium, ethacrynic acid, and a low external concentration of potassium on specific [3H]‐ouabain binding to human lymphocytes after incubation for 3 days.

The effects of serum, lithium, ethacrynic acid, and a low external concentration of potassium on specific [3H]‐ouabain binding to human lymphocytes after incubation for 3 days.

Na pump isoforms in human erythroid progenitor cells and mature erythrocytes

DOWN‐REGULATION OF THE SODIUM PUMP FOLLOWING CHRONIC EXPOSURE OF HeLa CELLS AND CHICK EMBRYO HEART CELLS TO OUABAIN

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