Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 1999 Nov;77(5):2602–2611. doi: 10.1016/S0006-3495(99)77094-3

Dynamics of oscillating erythrocyte doublets after electrofusion

M Baumann 1
PMCID: PMC1300534  PMID: 10545360

Abstract

Erythrocytes were electrofused with multiple rectangular voltage pulses to show an oscillatory movement, divided into swell phases and pump events. During each swell phase, which lasted from 0.5 s to more than 180 s, the fused cells' (doublets') volume increased by colloid osmotic swelling, and the membrane area was expanded until rupture. Membrane rupture initiated the pump event, where the doublets' volume and membrane area decreased with an almost exponential time course and time constants between 2 ms and 8 ms. Simultaneously, a portion of cytosolic hemoglobin solution was ejected into extracellular space ("jet"). Pump event time constants and swell phase durations decreased with rising chamber temperature, indicating that both parts of the oscillatory movements were determined by physical properties of membrane and liquids. Relative volume change developments express a gradual loss of membrane elasticity during the oscillation, decreasing the elastic forces stored in the membrane. Evidence is given that the first rupture causes a weakening of the membrane at the rupture site. Heat treatment up to 45 degrees C had a negligible effect on swell times, pump time constants, and relative volume changes. A heat treatment of 50 degrees C prevented oscillatory movements. The rupture location accorded with theories of potential induced membrane electropermeabilization.

Full Text

The Full Text of this article is available as a PDF (885.2 KB).

Selected References

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

  1. Akkaş N. Biomechanics of virus-to-cell and cell-to-cell fusion. J Biomed Eng. 1984 Oct;6(4):257–264. doi: 10.1016/0141-5425(84)90071-2. [DOI] [PubMed] [Google Scholar]
  2. Baumann M., Grebe R. Characteristics of the osmotically induced membrane rupture. Mol Membr Biol. 1998 Oct-Dec;15(4):193–201. doi: 10.3109/09687689709044321. [DOI] [PubMed] [Google Scholar]
  3. Baumann M., Sowers A. E. A mechanical cycling phenomenon in electrofused eryhrocytes. Mol Membr Biol. 1996 Apr-Jun;13(2):113–119. doi: 10.3109/09687689609160585. [DOI] [PubMed] [Google Scholar]
  4. Baumann M., Sowers A. E. Membrane skeleton involvement in cell fusion kinetics: a parameter that correlates with erythrocyte osmotic fragility. Biophys J. 1996 Jul;71(1):336–340. doi: 10.1016/S0006-3495(96)79229-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boal D. H. Computer simulation of a model network for the erythrocyte cytoskeleton. Biophys J. 1994 Aug;67(2):521–529. doi: 10.1016/S0006-3495(94)80511-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brandts J. F., Erickson L., Lysko K., Schwartz A. T., Taverna R. D. Calorimetric studies of the structural transitions of the human erythrocyte membrane. The involvement of spectrin in the A transition. Biochemistry. 1977 Jul 26;16(15):3450–3454. doi: 10.1021/bi00634a024. [DOI] [PubMed] [Google Scholar]
  7. Chernomordik L. V., Sowers A. E. Evidence that the spectrin network and a nonosmotic force control the fusion product morphology in electrofused erythrocyte ghosts. Biophys J. 1991 Nov;60(5):1026–1037. doi: 10.1016/S0006-3495(91)82140-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Evans E. A., Waugh R., Melnik L. Elastic area compressibility modulus of red cell membrane. Biophys J. 1976 Jun;16(6):585–595. doi: 10.1016/S0006-3495(76)85713-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Henszen M. M., Weske M., Schwarz S., Haest C. W., Deuticke B. Electric field pulses induce reversible shape transformation of human erythrocytes. Mol Membr Biol. 1997 Oct-Dec;14(4):195–204. doi: 10.3109/09687689709048182. [DOI] [PubMed] [Google Scholar]
  10. Hernandez L. D., Hoffman L. R., Wolfsberg T. G., White J. M. Virus-cell and cell-cell fusion. Annu Rev Cell Dev Biol. 1996;12:627–661. doi: 10.1146/annurev.cellbio.12.1.627. [DOI] [PubMed] [Google Scholar]
  11. Jay A. W., Canham P. B. Viscoelastic properties of the human red blood cell membrane. II. Area and volume of individual red cells entering a micropipette. Biophys J. 1977 Feb;17(2):169–178. doi: 10.1016/S0006-3495(77)85634-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lentz B. R. Polymer-induced membrane fusion: potential mechanism and relation to cell fusion events. Chem Phys Lipids. 1994 Sep 6;73(1-2):91–106. doi: 10.1016/0009-3084(94)90176-7. [DOI] [PubMed] [Google Scholar]
  13. Marsden N. V., Zade-Oppen A. M., Davies H. G. Jet expulsion of cellular contents from rad cells during photodynamic hemolysis. Ups J Med Sci. 1981;86(1):1–8. doi: 10.3109/03009738109179204. [DOI] [PubMed] [Google Scholar]
  14. Mehrle W., Hampp R., Zimmermann U. Electric pulse induced membrane permeabilization. Spatial orientation and kinetics of solute efflux in freely suspended and dielectrophoretically aligned plant mesophyll protoplasts. Biochim Biophys Acta. 1989 Jan 30;978(2):267–275. doi: 10.1016/0005-2736(89)90124-7. [DOI] [PubMed] [Google Scholar]
  15. Montané M. H., Dupille E., Alibert G., Teissié J. Induction of a long-lived fusogenic state in viable plant protoplasts permeabilized by electric fields. Biochim Biophys Acta. 1990 May 9;1024(1):203–207. doi: 10.1016/0005-2736(90)90227-f. [DOI] [PubMed] [Google Scholar]
  16. Morris S. J., Sarkar D. P., White J. M., Blumenthal R. Kinetics of pH-dependent fusion between 3T3 fibroblasts expressing influenza hemagglutinin and red blood cells. Measurement by dequenching of fluorescence. J Biol Chem. 1989 Mar 5;264(7):3972–3978. [PubMed] [Google Scholar]
  17. Müller G. H., Schmid-Schönbein H., Meiselman H. J. Development of viscoelasticity in heated hemoglobin solutions. Biorheology. 1992 Mar-Jun;29(2-3):203–216. doi: 10.3233/bir-1992-292-302. [DOI] [PubMed] [Google Scholar]
  18. Potter H. Electroporation in biology: methods, applications, and instrumentation. Anal Biochem. 1988 Nov 1;174(2):361–373. doi: 10.1016/0003-2697(88)90035-8. [DOI] [PubMed] [Google Scholar]
  19. Song L. Y., Ahkong Q. F., Georgescauld D., Lucy J. A. Membrane fusion without cytoplasmic fusion (hemi-fusion) in erythrocytes that are subjected to electrical breakdown. Biochim Biophys Acta. 1991 May 31;1065(1):54–62. doi: 10.1016/0005-2736(91)90010-6. [DOI] [PubMed] [Google Scholar]
  20. Sowers A. E. Low concentrations of macromolecular solutes significantly affect electrofusion yield in erythrocyte ghosts. Biochim Biophys Acta. 1990 Jun 27;1025(2):247–251. doi: 10.1016/0005-2736(90)90104-v. [DOI] [PubMed] [Google Scholar]
  21. Sowers A. E. Membrane skeleton restraint of surface shape change during fusion of erythrocyte membranes: evidence from use of osmotic and dielectrophoretic microforces as probes. Biophys J. 1995 Dec;69(6):2507–2516. doi: 10.1016/S0006-3495(95)80121-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sowers A. E. The long-lived fusogenic state induced in erythrocyte ghosts by electric pulses is not laterally mobile. Biophys J. 1987 Dec;52(6):1015–1020. doi: 10.1016/S0006-3495(87)83294-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Steubing R. W., Cheng S., Wright W. H., Numajiri Y., Berns M. W. Laser induced cell fusion in combination with optical tweezers: the laser cell fusion trap. Cytometry. 1991;12(6):505–510. doi: 10.1002/cyto.990120607. [DOI] [PubMed] [Google Scholar]
  24. Teissié J., Rols M. P. An experimental evaluation of the critical potential difference inducing cell membrane electropermeabilization. Biophys J. 1993 Jul;65(1):409–413. doi: 10.1016/S0006-3495(93)81052-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wu Y., Montes J. G., Sjodin R. A. Determination of electric field threshold for electrofusion of erythrocyte ghosts. Comparison of pulse-first and contact-first protocols. Biophys J. 1992 Mar;61(3):810–815. doi: 10.1016/S0006-3495(92)81885-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wu Y., Rosenberg J. D., Sowers A. E. Surface shape change during fusion of erythrocyte membranes is sensitive to membrane skeleton agents. Biophys J. 1994 Nov;67(5):1896–1905. doi: 10.1016/S0006-3495(94)80672-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wu Y., Sjodin R. A., Sowers A. E. Distinct mechanical relaxation components in pairs of erythrocyte ghosts undergoing fusion. Biophys J. 1994 Jan;66(1):114–119. doi: 10.1016/S0006-3495(94)80762-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Zade-Oppen A. M. Repetitive cell 'jumps' during hypotonic lysis of erythrocytes observed with a simple flow chamber. J Microsc. 1998 Oct;192(Pt 1):54–62. doi: 10.1046/j.1365-2818.1998.00402.x. [DOI] [PubMed] [Google Scholar]
  29. Zimmermann U. Electric field-mediated fusion and related electrical phenomena. Biochim Biophys Acta. 1982 Nov 30;694(3):227–277. doi: 10.1016/0304-4157(82)90007-7. [DOI] [PubMed] [Google Scholar]
  30. Zimmermann U., Vienken J. Electric field-induced cell-to-cell fusion. J Membr Biol. 1982;67(3):165–182. doi: 10.1007/BF01868659. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES