Laurdan fluorescence senses mechanical strain in the lipid bilayer membrane.

The precise molecular mechanisms by which cells transduce a mechanical stimulus into an intracellular biochemical response have not yet been established. Here, we show for the first time that the fluorescence emission of an environment-sensitive membrane probe Laurdan is modulated by mechanical strain of the lipid bilayer membrane. We have measured fluorescence emission of Laurdan in phospholipid vesicles of 30, 50, and 100 nm diameter to show that osmotically induced membrane tension leads to an increase in polarity (hydration depth) of the phospholipid bilayer interior. Our data indicate that the general polarization of Laurdan emission is linearly dependent on membrane tension. We also show that higher membrane curvature leads to higher hydration levels. We anticipate that the proposed method will facilitate future studies of mechanically induced changes in physical properties of lipid bilayer environment both in vitro and in vivo.

[1]  E Gratton,et al.  Quantitation of lipid phases in phospholipid vesicles by the generalized polarization of Laurdan fluorescence. , 1991, Biophysical journal.

[2]  F. W. Schneider,et al.  Influence of vesicle curvature on fluorescence relaxation kinetics of fluorophores. , 1994, Biophysical chemistry.

[3]  E. Gratton,et al.  Evidence for an increase in water concentration in bilayers after oxidative damage of phospholipids induced by ionizing radiation. , 1994, International journal of radiation biology.

[4]  E. Evans,et al.  Effect of chain length and unsaturation on elasticity of lipid bilayers. , 2000, Biophysical journal.

[5]  S. Strittmatter,et al.  P2Y1 purinergic receptors in sensory neurons: contribution to touch-induced impulse generation. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[6]  E. Evans,et al.  Water permeability and mechanical strength of polyunsaturated lipid bilayers. , 2000, Biophysical journal.

[7]  G. L. Jendrasiak The hydration of phospholipids and its biological significance , 1996 .

[8]  M. Bloom,et al.  Models of lipid-protein interactions in membranes. , 1993, Annual review of biophysics and biomolecular structure.

[9]  E. Gratton,et al.  Cholesterol modifies water concentration and dynamics in phospholipid bilayers: a fluorescence study using Laurdan probe. , 1994, Biophysical journal.

[10]  M. Paternostre,et al.  Laurdan solvatochromism: solvent dielectric relaxation and intramolecular excited-state reaction. , 1997, Biophysical journal.

[11]  Evans,et al.  Entropy-driven tension and bending elasticity in condensed-fluid membranes. , 1990, Physical review letters.

[12]  E. Evans,et al.  Osmotic properties of large unilamellar vesicles prepared by extrusion. , 1993, Biophysical journal.

[13]  J A Frangos,et al.  Modulation of GTPase activity of G proteins by fluid shear stress and phospholipid composition. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  P. Kinnunen,et al.  Phospholipase A2 as a mechanosensor. , 1995, Biophysical journal.

[15]  P. Chong,et al.  Interactions of Laurdan with phosphatidylcholine liposomes: a high pressure FTIR study. , 1993, Biochimica et biophysica acta.

[16]  J. Sadoshima,et al.  Autocrine release of angiotensin II mediates stretch-induced hypertrophy of cardiac myocytes in vitro , 1993, Cell.

[17]  J. Gier,et al.  The effect of chain length and lipid phase transitions on the selective permeability properties of liposomes. , 1975, Biochimica et biophysica acta.

[18]  Anthony G Lee,et al.  How lipids affect the activities of integral membrane proteins. , 2004, Biochimica et biophysica acta.

[19]  D. Discher,et al.  Bending Contributions to Hydration of Phospholipid and Block Copolymer Membranes: Unifying Correlations between Probe Fluorescence and Vesicle Thermoelasticity , 2001 .

[20]  Klaus Schulten,et al.  Lipid bilayer pressure profiles and mechanosensitive channel gating. , 2004, Biophysical journal.

[21]  Enrico Gratton,et al.  Laurdan and Prodan as Polarity-Sensitive Fluorescent Membrane Probes , 1998, Journal of Fluorescence.

[22]  F. Hallett,et al.  Optical changes in unilamellar vesicles experiencing osmotic stress. , 1996, Biophysical journal.

[23]  Boris Martinac,et al.  Physical principles underlying the transduction of bilayer deformation forces during mechanosensitive channel gating , 2002, Nature Structural Biology.

[24]  V A Parsegian,et al.  Osmotic stress, crowding, preferential hydration, and binding: A comparison of perspectives. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[25]  P. Kinnunen,et al.  Comparison of the effects of surface tension and osmotic pressure on the interfacial hydration of a fluid phospholipid bilayer. , 2003, Biophysical journal.

[26]  G. Klose Biomembranes, Physical Aspects , 1996 .

[27]  E Gratton,et al.  Phase fluctuation in phospholipid membranes revealed by Laurdan fluorescence. , 1990, Biophysical journal.