Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1981 Jan 1;88(1):51–56. doi: 10.1083/jcb.88.1.51

Intracellular univalent cations and the regulation of the BALB/c-3T3 cell cycle

PMCID: PMC2111729  PMID: 7204489

Abstract

Addition of serum to density-arrested BALB/c-3T3 cells causes a rapid increase in uptake of Na+ and K+, followed 12 h later by the onset of DNA synthesis. We explored the role of intracellular univalent cation concentrations in the regulation of BALB/c-3T3 cell growth by serum growth factors. As cells grew to confluence, intracellular Na+ and K+ concentrations ([Na+]i and [K+]i) fell from 40 and 180 to 15 and 90 mmol/liter, respectively. Stimulation of growth of density-inhibited cells by the addition of serum growth factors increased [Na]i by 30% and [K+]i by 13-25% in early G0/G1, resulting in an increase in total univalent cation concentration. Addition of ouabain to stimulated cells resulted in a concentration-dependent steady decrease in [K+]i and increase in [Na+]i. Ouabain (100 microM) decreased [K+]i to approximately 60 mmol/liter by 12 h, and also prevented the serum- stimulated increase in 86Rb+ uptake. However, 100 microM ouabain did not inhibit DNA synthesis. A time-course experiment was done to determine the effect of 100 microM ouabain on [K+]i throughout G0/G1 and S phase. The addition of serum growth factors to density-inhibited cells stimulated equal rates of entry into the S phase in the presence or absence of 100 microM ouabain. However, in the presence of ouabain, there was a decrease in [K+]i. Therefore, an increase in [K+]i is not required for entry into S phase; serum growth factors do not regulate cell growth by altering [K+]i. The significance of increased total univalent cation concentration is discussed.

Full Text

The Full Text of this article is available as a PDF (637.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allen W. R., Nilsen-Hamilton M., Hamilton R. T., Gospodarowicz D. Serum-dependent regulation of alpha-aminoisobutyric acid uptake in bovine granulosa cells. J Cell Physiol. 1979 Mar;98(3):491–502. doi: 10.1002/jcp.1040980308. [DOI] [PubMed] [Google Scholar]
  2. Carrasco L., Smith A. E. Sodium ions and the shut-off of host cell protein synthesis by picornaviruses. Nature. 1976 Dec 23;264(5588):807–809. doi: 10.1038/264807a0. [DOI] [PubMed] [Google Scholar]
  3. Cook J. S., Vaughan G. L., Proctor W. R., Brake E. T. Interaction of two mechanisms regulating alkali cations in HeLa cells. J Cell Physiol. 1975 Aug;86(1):59–70. doi: 10.1002/jcp.1040860108. [DOI] [PubMed] [Google Scholar]
  4. Frantz C. N., Stiles C. D., Pledger W. J., Scher C. D. Effect of ouabain on growth regulation by serum components in Balb/c-3T3 cells: inhibition of entry into S phase by decreased protein synthesis. J Cell Physiol. 1980 Dec;105(3):439–448. doi: 10.1002/jcp.1041050308. [DOI] [PubMed] [Google Scholar]
  5. Hershko A., Mamont P., Shields R., Tomkins G. M. "Pleiotypic response". Nat New Biol. 1971 Aug;232(33):206–211. [PubMed] [Google Scholar]
  6. Holian A., Deutsch C. J., Holian S. K., Daniele R. P., Wilson D. F. Lymphocyte response to phytohemagglutinin: intracellular volume and intracellular [K+]. J Cell Physiol. 1979 Jan;98(1):137–144. doi: 10.1002/jcp.1040980115. [DOI] [PubMed] [Google Scholar]
  7. Holley R. W. Control of growth of mammalian cells in cell culture. Nature. 1975 Dec 11;258(5535):487–490. doi: 10.1038/258487a0. [DOI] [PubMed] [Google Scholar]
  8. Kletzien R. F., Pariza M. W., Becker J. E., Potter V. R. A method using 3-O-methyl-D-glucose and phloretin for the determination of intracellular water space of cells in monolayer culture. Anal Biochem. 1975 Oct;68(2):537–544. doi: 10.1016/0003-2697(75)90649-1. [DOI] [PubMed] [Google Scholar]
  9. Koch K. S., Leffert H. L. Increased sodium ion influx is necessary to initiate rat hepatocyte proliferation. Cell. 1979 Sep;18(1):153–163. doi: 10.1016/0092-8674(79)90364-7. [DOI] [PubMed] [Google Scholar]
  10. Ledbetter M. L., Lubin M. Control of protein synthesis in human fibroblasts by intracellular potassium. Exp Cell Res. 1977 Mar 15;105(2):223–236. doi: 10.1016/0014-4827(77)90120-3. [DOI] [PubMed] [Google Scholar]
  11. McKeehan W. L., McKeehan K. A. Serum factors modify the cellular requirement for Ca2+, K+, Mg2+, phosphate ions, and 2-oxocarboxylic acids for multiplication of normal human fibroblasts. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3417–3421. doi: 10.1073/pnas.77.6.3417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mendoza S. A., Wigglesworth N. M., Pohjanpelto P., Rozengurt E. Na entry and Na-K pump activity in murine, hamster, and human cells--effect of monensin, serum, platelet extract, and viral transformation. J Cell Physiol. 1980 Apr;103(1):17–27. doi: 10.1002/jcp.1041030104. [DOI] [PubMed] [Google Scholar]
  13. Moscatelli D., Sanui H., Rubin A. H. Effects of depletion of K+, Na+, or Ca2+ on DNA synthesis and cell cation content in chick embryo fibroblasts. J Cell Physiol. 1979 Oct;101(1):117–128. doi: 10.1002/jcp.1041010114. [DOI] [PubMed] [Google Scholar]
  14. Pledger W. J., Stiles C. D., Antoniades H. N., Scher C. D. Induction of DNA synthesis in BALB/c 3T3 cells by serum components: reevaluation of the commitment process. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4481–4485. doi: 10.1073/pnas.74.10.4481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rozengurt E., Heppel L. A. Serum rapidly stimulates ouabain-sensitive 86-RB+ influx in quiescent 3T3 cells. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4492–4495. doi: 10.1073/pnas.72.11.4492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Segel G. B., Simon W., Lichtman M. A. Regulation of sodium and potassium transport in phytohemagglutinin-stimulated human blood lymphocytes. J Clin Invest. 1979 Sep;64(3):834–841. doi: 10.1172/JCI109531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Smith J. A., Martin L. Do cells cycle? Proc Natl Acad Sci U S A. 1973 Apr;70(4):1263–1267. doi: 10.1073/pnas.70.4.1263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith J. B., Rozengurt E. Serum stimulates the Na+,K+ pump in quiescent fibroblasts by increasing Na+ entry. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5560–5564. doi: 10.1073/pnas.75.11.5560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Spaggiare S., Wallach M. J., Tupper J. T. Potassium transport in normal and transformed mouse 3T3 cells. J Cell Physiol. 1976 Nov;89(3):403–416. doi: 10.1002/jcp.1040890306. [DOI] [PubMed] [Google Scholar]
  20. Tupper J. T., Zografos L. Effect of imposed serum deprivation on growth of the mouse 3T3 cell. Dissociation from changes in potassium ion transport as measured from [86Rb)rubidium ion uptake. Biochem J. 1978 Sep 15;174(3):1063–1065. doi: 10.1042/bj1741063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tupper J. T., Zorgniotti F., Mills B. Potassium transport and content during G1 and S phase following serum stimulation of 3T3 cells. J Cell Physiol. 1977 Jun;91(3):429–440. doi: 10.1002/jcp.1040910313. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES