Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Hypotonically induced changes in the plasma membrane of cultured mammalian cells

Summary

Cells from three cell lines were electrorotated in media of osmotic strengths from 330 mOsm to 60 mOsm. From the field-frequency dependence of the rotation speed, the passive electrical properties of the surfaces were deduced. In all cases, the area-specific membrane capacitance (C m) decreased with osmolality. At 280 mOsm (iso-osmotic), SP2 (mouse myeloma) and G8 (hybridoma) cells had C mvalues of 1.01 ± 0.04 μF/cm2 and 1.09 ± 0.03 μF/cm2, respectively, whereas dispase-treated L-cells (sarcoma fibroblasts) exhibited C m=2.18±0.10/μF/cm2. As the osmolality was reduced, the C mreached a well-defined minimum at 150 mOsm (SP2) or 180 mOsm (G8). Further reduction in osmolality gave a 7% increase in C m, after which a plateau close to 0.80μF/cm22was reached. However, the whole-cell capacities increased about twofold from 200 mOsm to 60 mOsm. L-cells showed very little change in C mbetween 280 mOsm and 150 mOsm, but below 150 mOsm the C mdecreased rapidly. The changes in C mcorrelate well with the swelling of the cells assessed by means of van't Hoff plots. The apparent membrane conductance (including the effect of surface conductance) decreased with C m, but then increased again instead of exhibiting a plateau. The rotation speed of the cells increased as the osmolality was lowered, and eventually attained almost the theoretical value. All measurements indicate that hypo-osmotically stressed cells obtain the necessary membrane area by using material from microvilli. However, below about 200 mOsm the whole-cell capacities indicate the progressive incorporation of “extra” membrane into the cell surface.

This is a preview of subscription content, log in to check access.

References

  1. 1.

    Ahkong, Q.F., Lucy, J.A. 1986. Osmotic forces in artificially induced cell fusion. Biochim. Biophys. Acta 858:206–216

  2. 2.

    Arnold, W.M. 1988. Analysis of optimum electro-rotation technique. Ferroelectrics 86:225–244

  3. 3.

    Arnold, W.M., Gessner, A.G., Zimmermann, U. 1993. Dielectric measurements on electromanipulation media. Biochim. Biophys. Acta 1157: (in press)

  4. 4.

    Arnold, W.M., Klarmann, B.G., Sukhorukov, V.L., Zimmermann, U. 1992. Membrane accommodation in hypo-osmotically-treated, and giant electrofused cells. Biochem. Soc. (Lond.) Transactions 20:120S

  5. 5.

    Arnold, W.M., Schmutzler, R.K., Al-Hasani, S., Krebs, D., Zimmermann, U. 1989. Differences in membrane properties between unfertilised and fertilised single rabbit oocytes demonstrated by electro-rotation. Comparison with cells from early embryos. Biochim. Biophys. Acta 979:142–146

  6. 6.

    Arnold, W.M., Schwan, H.P., Zimmermann, U. 1987. Surface conductance and other properties of latex particles measured by electrorotation. J. Phys. Chem. 91:5093–5098

  7. 7.

    Arnold, W.M., Zimmermann, U. 1982. Rotation of an isolated cell in a rotating electric field. Naturwissenschaften 69:297

  8. 8.

    Arnold, W.M., Zimmermann, U. 1983. Patent application, official designation P3325 843.0, received at the Patent Office, FRG, July 18, 1983

  9. 9.

    Arnold, W.M., Zimmermann, U. 1988. Electro-rotation: development of a technique for dielectric measurements on individual cells and particles. J. Electrostatics 21:151–191

  10. 10.

    Arnold, W.M., Zimmermann, U. 1989. Measurements of dielectric properties of single cells or other particles using direct observation of electro-rotation. In: Proceedings of The First International Conference on Low Cost Experiments in Biophysics, Cairo, 18–20 December, 1989. pp. 1–13

  11. 11.

    Asami, K., Takahashi, Y., Takashima, S. 1989. Dielectric properties of mouse lymphocytes and erythrocytes. Biochim. Biophys. Acta 1010:49–55

  12. 12.

    Asami, K., Takahashi, Y., Takashima, S. 1990. Frequency domain analysis of membrane capacitance of cultured cells (HeLa and myeloma) using the micropipette technique. Biophys. J. 58:143–148

  13. 13.

    Chizmadzhev, Yu.A., Kuzmin, P.I., Pastushenko, V.Ph. 1985. Theory of the dielectrophoresis of vesicles and cells. Biologicheskie Membrany 2:1147–1161 (in Russian)

  14. 14.

    CRC 1988. CRC Handbook of Chemistry and Physics, R.C. Weast, editor. CRC Press, Boca Raton, FL

  15. 15.

    Earle, W.R. 1943. Production of malignancy in vitro. IV. The mouse fibroblast cultures and changes seen in the living cells. J. Nat. Cancer Inst. 4:165–212

  16. 16.

    Foung, S., Perkins, S., Kafadar, K., Gessner, P., Zimmermann, U. 1990. Development of microfusion techniques to generate human hybridomas. J. Immunol. Meth. 134:35–42

  17. 17.

    Freitag, R., Schügerl, K., Arnold, W.M., Zimmermann, U. 1989. The effect of osmotic and mechanical stresses and enzymatic digestion on the electro-rotation of insect cells (Spodoptera frugiperda). J. Biotechnol. 11:325–336

  18. 18.

    Fuhr, G., Arnold, W.M., Hagedorn, R., Müller, T., Benecke, W., Wagner, B., Zimmermann, U. 1992. Levitation, holding, and rotation of cells within traps made by high frequency fields. Biochim. Biophys. Acta 1108:215–223

  19. 19.

    Fuhr, G., Glaser, R., Hagedorn, R. 1986. Rotation of dielectrics in a rotating electric high-frequency field. Biophys. J. 69:395–402

  20. 20.

    Fuhr, G., Hagedorn, R. 1988. Grundlagen der Elektrorotation. In: Colloquia Pflanzenphysiologie. H. Göring und P. Hoffmann, editors, Nr. 11. Humboldt-Universität zu Berlin, Berlin

  21. 21.

    Fuhr, G., Kuzmin, P.I. 1986. Behavior of cells in rotating electric fields with account to surface charges and cell structures. Biophys. J. 50:789–795

  22. 22.

    Glaser, R., Fuhr, G., Gimsa, J. 1983. Rotation of erythrocytes, plant cells, and protoplasts in an outside rotating electric field. Studia Biophysica 96:11–20

  23. 23.

    Harris, C.M., Kell, D.B. 1985. On the dielectrically observable consequences of the diffusional motions of lipids and proteins in membranes. 2. Experiments with microbial cells, protoplasts and membrane vesicles. Eur. Biophys. J. 13:11–24

  24. 24.

    Hu, X., Arnold, W.M., Zimmermann, U. 1990. Alterations in the electrical properties of T and B lymphocyte membranes induced by mitogenic stimulation. Activation monitored by electro-rotation of single cells. Biochim. Biophys. Acta 1021:191–200

  25. 25.

    Kaler, K.V.I.S., Jones, T.B. 1990. Dielectrophoretic spectra of single cells determined by feedback controlled levitation. Biophys. J. 57:173–182

  26. 26.

    Kell, D.B., Harris, C.M. 1985. On the dielectrically observable consequences of the diffusional motions of lipids and proteins in membranes. 1. Theory and overview. Eur. Biophys. J. 12:181–197

  27. 27.

    Klöck, G., Zimmermann, U. 1990. Facilitated electrofusion of vacuolated x evacuolated oat mesophyll protoplasts in hypo-osmotic media after alignment with an alternating field of modulated strength. Biochim. Biophys. Acta 1025:87–93

  28. 28.

    Knutton, S., Jackson, D., Graham, J.M., Micklem, K.J., Pasternak, C.A. 1976. Microvilli and cell swelling. Nature 262:52–54

  29. 29.

    Lamb, H. 1906. Hydrodynamics, 3rd edn., Article 322. Cambridge University, Cambridge, UK

  30. 30.

    Mela, M., Eskelinen, S. 1984. Normal and homogeneous red blood cell populations over a wide range of hyper-isohypotonic media. III. Corrected volumes in Coulter Counter measurements. Acta Physiol. Scand. 122:515–525

  31. 31.

    Pethig, R. 1979. Dielectric and Electronic Properties of Biological Materials. John Wiley and Sons, Chichester

  32. 32.

    Pethig, R. 1991. Application of A.C. electrical fields to the manipulation and characterisation of cells. In: Automation in Biotechnology. I. Karube, editor. pp. 159–185. Elsevier Science Publishers B.V.

  33. 33.

    Sauer, F.A., Schlögl, R.W. 1985. Torques exerted on cylinders and spheres by external electromagnetic fields. A contribution to the theory of induced cell rotation, In: Interaction Between Electromagnetic Fields and Cells. A. Chiabrera, C. Nicolini and H.P. Schwan, editors. pp. 203–251. Plenum, New York

  34. 34.

    Schmitt, J.J., Zimmermann, U. 1989. Enhanced hybridoma production by electrofusion in strongly hypo-osmolar solutions. Biochim. Biophys. Acta 983:42–50

  35. 35.

    Schwan, H.P. 1985. Dielectric properties of the cell surface and electric field effects on cells. Studia Biophysica 110:13–18

  36. 36.

    Schwan, H.P. 1988. Dielectric spectroscopy and electro-rotation of biological cells. Ferroelectrics 86:205–223

  37. 37.

    Schwan, H.P. 1989. Dielectrophoresis and rotation of cells. In: Electroporation and Electrofusion in Cell Biology. E. Neuman, A.E. Sowers and C.A. Jordan, editors. pp. 3–21. Plenum, New York

  38. 38.

    Shulman, M., Wilde, C.D., Köhler, G. 1978. A better cell line for marking hybridomas secreting specific antibodies. Nature 276: 269–270

  39. 39.

    Steponkus, P.L., Lynch, D.V. 1989. Freeze/thaw-induced destabilization of the plasma membrane and the effects of cold acclimation. J. Bioenerg. Biomembr. 21:21–41

  40. 40.

    Zimmermann, U., Arnold, W.M. 1983. The interpretation and use of the rotation of biological cells. In: Coherent Excitations in Biological Systems. H. Fröhlich and F. Kremer, editors. pp. 211–221. Springer-Verlag, Berlin

  41. 41.

    Zimmermann, U., Gessner, P., Schnettler, R., Perkins, S., Foung, S.K.H. 1990. Efficient hybridization of mouse-human cell lines by means of hypo-osmolar electrofusion. J. Immunol. Meth. 134:43–50

Download references

Author information

Additional information

We thank Mr. B.G. Klarmann for his help with the measurements. This work was supported by grants of the DFG (SFB 176 B5 to U.Z. and W.M.A.) and of the BMFT (DARA 50 WB 9212 to U.Z.). We also thank the Umweltbundesamt, Berlin, for support enabling the construction of some of the rotation generators used in this work.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Sukhorukov, V.L., Arnold, W.M. & Zimmermann, U. Hypotonically induced changes in the plasma membrane of cultured mammalian cells. J. Membarin Biol. 132, 27–40 (1993). https://doi.org/10.1007/BF00233049

Download citation

Key Words

  • membrane stress
  • osmotic pressure
  • membrane
  • conductivity
  • membrane capacity
  • electrorotation