TMA-DPH as Specific Plasma Membrane Fluidity Probe for Intact Cells and its Limitation

  • M. Kubina
  • J. G. Kuhry
  • G. Duportail
  • G. Coupin
  • C. Bronner
  • P. Poindron
  • G. Laustriat
Part of the NATO ASI Series book series (NSSA, volume 116)

Abstract

Fluorescence anisotropy measurements to determine membrane fluidity have been generally performed with diphenylhexatriene (DPH) on purified membrane samples. This probe could not be used satisfactorily with intact cells, because it did not present any specificity for a particular type of membrane and labelled all the hydrophobic regions of the cell. We have recently focussed our attention on the cationic derivative of DPH, trimethylamino-diphenylhexatriene (TMA-DPH). This molecule displays excellent photophysical properties in hydrophobic media: lifetime 6 ns (DPH 8 ns), quantum yield 0.55 (DPH 0.75), and is not fluorescent in water.12 In comparison with DPH and the other fluorescent probes previously used in membrane fluidity determinations it was given evidence3,4 by fluorescence micrography and by fluorescence intensity measurements, to have two particularly favorable features :
  1. (i)

    it is incorporated in membranes very rapidly

     
  2. (ii)

    when it is interacted with intact cells, it is retained specifically in the plasma membranes for periods of typically 30 min. before being internalized.

     

Keywords

Anisotropy Phenyl Interferon Histamine Noradrenalin 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    F.G. Prendergast, R.P. Hayland and P.J. Callahan, 1-(4-(trimethylaminophenyl)-6-phenylhexa-1,3,5-triene: synthesis, fluorescence properties and use as a fluorescence probe of lipid bilayers, Biochemistry 20: 7333–7337 (1981).PubMedCrossRefGoogle Scholar
  2. 2.
    M.N. Cranney, R.B. Cundall, G.R. Jones, J.T. Richards, and E.W. Thomas, Fluorescence lifetime and quenching studies on some interesting diphenylhexatriene membrane probes, Biochim. Biophys. Acta 731: 387–396 (1983).CrossRefGoogle Scholar
  3. 3.
    J.G. Kuhry, P. Fonteneau, G. Duportail, C. Maechling, and G. Laustriat, TMA-DPH: A suitable fluorescence polarization probe for specific plasma membrane fluidity studies in intact living cells, Cell Biophys. 5: 129–140 (1983).PubMedGoogle Scholar
  4. 4.
    J.G. Kuhry, G. Duportail, C. Bronner, and G. Laustriat, Plasma membrane fluidity measurements on whole living cells by fluorescence anisotropy of trimethylammoniumdiphenylhexatriene, Biochim. Biophys. Acta 845: 60–67 (1985).PubMedCrossRefGoogle Scholar
  5. 5.
    J.G. Kuhry, C. Bronner, M. Amelia, and Y. Landry, Plasma membrane fluidity studies by fluorescence polarization in rat mast cells stimulated by compound 48/80, Agents Actions 16: 109–112 (1985).PubMedCrossRefGoogle Scholar
  6. 6.
    A. Deguercy, J. Schrevel, G. Duportail, G. Laustriat, and J.G. Kuhry, Membrane fluidity changes in P. Berghei infected erythrocytes, Investigated with a specific plasma membrane fluidity probe, Biochem. Int., In press.Google Scholar
  7. 7.
    C. Mutet, G. Duportail, G. Cremel, and A. Waksman, Increase of the fluidity of the lipid bilayer of the inner mitochondrial membrane by succinate and phenylsuccinate: a study by EPR and fluorescence, Biochim. Biophys. Res. Comm. 119: 854–859 (1984).CrossRefGoogle Scholar
  8. 8.
    C. Bronner, J.G. Kuhry, and Y. Landry, A fluorescent hydrophobic probe allows to monitor mast cell exocytosis, Agents Actions, In press (1986).Google Scholar
  9. 9.
    M.G. Burgess, F. Giraud, J. Poggioli, and M.2Claret, a-Adrenergically mediated changes in membrane fluidity and Ca binding in isolated rat liver plasma membranes, Biochim. Biophys. Acta 731: 387–396 (1983).PubMedCrossRefGoogle Scholar
  10. 10.
    J.G. Kuhry, P. Poindron, and G. Laustriat, Evidence for early fluidity changes in the plasma membranes of interferon treated L cells, from fluorescence anisotropy data, Biochem. Biophys. Res. Comm. 110: 88–95 (1983).PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1986

Authors and Affiliations

  • M. Kubina
    • 1
  • J. G. Kuhry
    • 1
  • G. Duportail
    • 1
  • G. Coupin
    • 1
    • 2
  • C. Bronner
    • 1
    • 3
  • P. Poindron
    • 1
    • 2
  • G. Laustriat
    • 1
  1. 1.Laboratoire de Biophysique, U.A. 491 du CNRSUniversité Louis Pasteur, Faculté de PharmacieStrasbourg CédexFrance
  2. 2.Laboratoire de VirologieUniversité Louis Pasteur, Faculté de PharmacieStrasbourg CédexFrance
  3. 3.Laboratoire d’AllergopharmacologieUniversité Louis Pasteur, Faculté de PharmacieStrasbourg CédexFrance

Personalised recommendations