Abstract
Optical tweezers were used to characterize the mechanical properties of the outer hair cell (OHC) plasma membrane by pulling tethers with 4.5-microm polystyrene beads. Tether formation force and tether force were measured in static and dynamic conditions. A greater force was required for tether formations from OHC lateral wall (499 +/- 152 pN) than from OHC basal end (142 +/- 49 pN). The difference in the force required to pull tethers is consistent with an extensive cytoskeletal framework associated with the lateral wall known as the cortical lattice. The apparent plasma membrane stiffness, estimated under the static conditions by measuring tether force at different tether length, was 3.71 pN/microm for OHC lateral wall and 4.57 pN/microm for OHC basal end. The effective membrane viscosity was measured by pulling tethers at different rates while continuously recording the tether force, and estimated in the range of 2.39 to 5.25 pN x s/microm. The viscous force most likely results from the viscous interactions between plasma membrane lipids and the OHC cortical lattice and/or integral membrane proteins. The information these studies provide on the mechanical properties of the OHC lateral wall is important for understanding the mechanism of OHC electromotility.
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- Arima T., Kuraoka A., Toriya R., Shibata Y., Uemura T. Quick-freeze, deep-etch visualization of the 'cytoskeletal spring' of cochlear outer hair cells. Cell Tissue Res. 1991 Jan;263(1):91–97. doi: 10.1007/BF00318403. [DOI] [PubMed] [Google Scholar]
- Ashkin A., Dziedzic J. M., Yamane T. Optical trapping and manipulation of single cells using infrared laser beams. Nature. 1987 Dec 24;330(6150):769–771. doi: 10.1038/330769a0. [DOI] [PubMed] [Google Scholar]
- Ashkin A. Forces of a single-beam gradient laser trap on a dielectric sphere in the ray optics regime. Biophys J. 1992 Feb;61(2):569–582. doi: 10.1016/S0006-3495(92)81860-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ashkin A., Schütze K., Dziedzic J. M., Euteneuer U., Schliwa M. Force generation of organelle transport measured in vivo by an infrared laser trap. Nature. 1990 Nov 22;348(6299):346–348. doi: 10.1038/348346a0. [DOI] [PubMed] [Google Scholar]
- Ashmore J. F. A fast motile response in guinea-pig outer hair cells: the cellular basis of the cochlear amplifier. J Physiol. 1987 Jul;388:323–347. doi: 10.1113/jphysiol.1987.sp016617. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Block S. M., Goldstein L. S., Schnapp B. J. Bead movement by single kinesin molecules studied with optical tweezers. Nature. 1990 Nov 22;348(6299):348–352. doi: 10.1038/348348a0. [DOI] [PubMed] [Google Scholar]
- Bo L., Waugh R. E. Determination of bilayer membrane bending stiffness by tether formation from giant, thin-walled vesicles. Biophys J. 1989 Mar;55(3):509–517. doi: 10.1016/S0006-3495(89)82844-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brownell W. E., Bader C. R., Bertrand D., de Ribaupierre Y. Evoked mechanical responses of isolated cochlear outer hair cells. Science. 1985 Jan 11;227(4683):194–196. doi: 10.1126/science.3966153. [DOI] [PubMed] [Google Scholar]
- Brownell W. E. Outer hair cell electromotility and otoacoustic emissions. Ear Hear. 1990 Apr;11(2):82–92. doi: 10.1097/00003446-199004000-00003. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brownell W. E., Spector A. A., Raphael R. M., Popel A. S. Micro- and nanomechanics of the cochlear outer hair cell. Annu Rev Biomed Eng. 2001;3:169–194. doi: 10.1146/annurev.bioeng.3.1.169. [DOI] [PubMed] [Google Scholar]
- Dai J., Sheetz M. P. Cell membrane mechanics. Methods Cell Biol. 1998;55:157–171. [PubMed] [Google Scholar]
- Dai J., Sheetz M. P. Mechanical properties of neuronal growth cone membranes studied by tether formation with laser optical tweezers. Biophys J. 1995 Mar;68(3):988–996. doi: 10.1016/S0006-3495(95)80274-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dai J., Sheetz M. P. Membrane tether formation from blebbing cells. Biophys J. 1999 Dec;77(6):3363–3370. doi: 10.1016/S0006-3495(99)77168-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dallos P., Corey M. E. The role of outer hair cell motility in cochlear tuning. Curr Opin Neurobiol. 1991 Aug;1(2):215–220. doi: 10.1016/0959-4388(91)90081-h. [DOI] [PubMed] [Google Scholar]
- Dieler R., Shehata-Dieler W. E., Brownell W. E. Concomitant salicylate-induced alterations of outer hair cell subsurface cisternae and electromotility. J Neurocytol. 1991 Aug;20(8):637–653. doi: 10.1007/BF01187066. [DOI] [PubMed] [Google Scholar]
- Evans E. A., Hochmuth R. M. Membrane viscoelasticity. Biophys J. 1976 Jan;16(1):1–11. doi: 10.1016/S0006-3495(76)85658-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Evans E. A., Hochmuth R. M. Membrane viscoplastic flow. Biophys J. 1976 Jan;16(1):13–26. doi: 10.1016/S0006-3495(76)85659-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Finer J. T., Simmons R. M., Spudich J. A. Single myosin molecule mechanics: piconewton forces and nanometre steps. Nature. 1994 Mar 10;368(6467):113–119. doi: 10.1038/368113a0. [DOI] [PubMed] [Google Scholar]
- Fluhler E., Burnham V. G., Loew L. M. Spectra, membrane binding, and potentiometric responses of new charge shift probes. Biochemistry. 1985 Oct 8;24(21):5749–5755. doi: 10.1021/bi00342a010. [DOI] [PubMed] [Google Scholar]
- Hochmuth F. M., Shao J. Y., Dai J., Sheetz M. P. Deformation and flow of membrane into tethers extracted from neuronal growth cones. Biophys J. 1996 Jan;70(1):358–369. doi: 10.1016/S0006-3495(96)79577-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hochmuth R. M., Mohandas N., Blackshear P. L., Jr Measurement of the elastic modulus for red cell membrane using a fluid mechanical technique. Biophys J. 1973 Aug;13(8):747–762. doi: 10.1016/S0006-3495(73)86021-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holley M. C., Kalinec F., Kachar B. Structure of the cortical cytoskeleton in mammalian outer hair cells. J Cell Sci. 1992 Jul;102(Pt 3):569–580. doi: 10.1242/jcs.102.3.569. [DOI] [PubMed] [Google Scholar]
- Hwang W. C., Waugh R. E. Energy of dissociation of lipid bilayer from the membrane skeleton of red blood cells. Biophys J. 1997 Jun;72(6):2669–2678. doi: 10.1016/S0006-3495(97)78910-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Iwasa K. H. A membrane motor model for the fast motility of the outer hair cell. J Acoust Soc Am. 1994 Oct;96(4):2216–2224. doi: 10.1121/1.410094. [DOI] [PubMed] [Google Scholar]
- Iwasa K. H. Effect of stress on the membrane capacitance of the auditory outer hair cell. Biophys J. 1993 Jul;65(1):492–498. doi: 10.1016/S0006-3495(93)81053-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuo S. C., Sheetz M. P. Force of single kinesin molecules measured with optical tweezers. Science. 1993 Apr 9;260(5105):232–234. doi: 10.1126/science.8469975. [DOI] [PubMed] [Google Scholar]
- Liang H., Vu K. T., Krishnan P., Trang T. C., Shin D., Kimel S., Berns M. W. Wavelength dependence of cell cloning efficiency after optical trapping. Biophys J. 1996 Mar;70(3):1529–1533. doi: 10.1016/S0006-3495(96)79716-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neuman K. C., Chadd E. H., Liou G. F., Bergman K., Block S. M. Characterization of photodamage to Escherichia coli in optical traps. Biophys J. 1999 Nov;77(5):2856–2863. doi: 10.1016/S0006-3495(99)77117-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nguyen T. V., Brownell W. E. Contribution of membrane cholesterol to outer hair cell lateral wall stiffness. Otolaryngol Head Neck Surg. 1998 Jul;119(1):14–20. doi: 10.1016/S0194-5998(98)70167-6. [DOI] [PubMed] [Google Scholar]
- Oghalai J. S., Patel A. A., Nakagawa T., Brownell W. E. Fluorescence-imaged microdeformation of the outer hair cell lateral wall. J Neurosci. 1998 Jan 1;18(1):48–58. doi: 10.1523/JNEUROSCI.18-01-00048.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pollice P. A., Brownell W. E. Characterization of the outer hair cell's lateral wall membranes. Hear Res. 1993 Nov;70(2):187–196. doi: 10.1016/0378-5955(93)90157-v. [DOI] [PubMed] [Google Scholar]
- Raphael R. M., Popel A. S., Brownell W. E. A membrane bending model of outer hair cell electromotility. Biophys J. 2000 Jun;78(6):2844–2862. doi: 10.1016/S0006-3495(00)76827-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Raucher D., Sheetz M. P. Characteristics of a membrane reservoir buffering membrane tension. Biophys J. 1999 Oct;77(4):1992–2002. doi: 10.1016/S0006-3495(99)77040-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Santos-Sacchi J. Harmonics of outer hair cell motility. Biophys J. 1993 Nov;65(5):2217–2227. doi: 10.1016/S0006-3495(93)81247-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shao J. Y., Hochmuth R. M. Micropipette suction for measuring piconewton forces of adhesion and tether formation from neutrophil membranes. Biophys J. 1996 Nov;71(5):2892–2901. doi: 10.1016/S0006-3495(96)79486-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sheetz M. P., Painter R. G., Singer S. J. Biological membranes as bilayer couples. III. Compensatory shape changes induced in membranes. J Cell Biol. 1976 Jul;70(1):193–203. doi: 10.1083/jcb.70.1.193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sheetz M. P., Singer S. J. Biological membranes as bilayer couples. A molecular mechanism of drug-erythrocyte interactions. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4457–4461. doi: 10.1073/pnas.71.11.4457. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sit P. S., Spector A. A., Lue A. J., Popel A. S., Brownell W. E. Micropipette aspiration on the outer hair cell lateral wall. Biophys J. 1997 Jun;72(6):2812–2819. doi: 10.1016/S0006-3495(97)78923-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith S. B., Cui Y., Bustamante C. Overstretching B-DNA: the elastic response of individual double-stranded and single-stranded DNA molecules. Science. 1996 Feb 9;271(5250):795–799. doi: 10.1126/science.271.5250.795. [DOI] [PubMed] [Google Scholar]
- Spector A. A., Brownell W. E., Popel A. S. Analysis of the micropipet experiment with the anisotropic outer hair cell wall. J Acoust Soc Am. 1998 Feb;103(2):1001–1006. doi: 10.1121/1.421216. [DOI] [PubMed] [Google Scholar]
- Spector A. A., Brownell W. E., Popel A. S. Estimation of elastic moduli and bending stiffness of the anisotropic outer hair cell wall. J Acoust Soc Am. 1998 Feb;103(2):1007–1011. doi: 10.1121/1.421217. [DOI] [PubMed] [Google Scholar]
- Waugh R. E., Bauserman R. G. Physical measurements of bilayer-skeletal separation forces. Ann Biomed Eng. 1995 May-Jun;23(3):308–321. doi: 10.1007/BF02584431. [DOI] [PubMed] [Google Scholar]
- Waugh R. E., Hochmuth R. M. Mechanical equilibrium of thick, hollow, liquid membrane cylinders. Biophys J. 1987 Sep;52(3):391–400. doi: 10.1016/S0006-3495(87)83227-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Waugh R. E., Song J., Svetina S., Zeks B. Local and nonlocal curvature elasticity in bilayer membranes by tether formation from lecithin vesicles. Biophys J. 1992 Apr;61(4):974–982. doi: 10.1016/S0006-3495(92)81904-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Waugh R. E. Surface viscosity measurements from large bilayer vesicle tether formation. I. Analysis. Biophys J. 1982 Apr;38(1):19–27. doi: 10.1016/S0006-3495(82)84526-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Waugh R. E. Surface viscosity measurements from large bilayer vesicle tether formation. II. Experiments. Biophys J. 1982 Apr;38(1):29–37. doi: 10.1016/S0006-3495(82)84527-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zahn M., Seeger S. Optical tweezers in pharmacology. Cell Mol Biol (Noisy-le-grand) 1998 Jul;44(5):747–761. [PubMed] [Google Scholar]