Skip to main content
Log in

Alterations in membrane fluidity during keratinocyte differentiation measured by fluorescence polarization

  • Short Communications
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Summary

The epidermis shows a distinctive pattern of differentiation wherein keratinocytes proliferate in the basal cell layer and mature into spinous and granular cells. Using a discontinuous density-gradient centrifugation method, guinea-pig keratinocytes were separated into high (HDF), intermediate (IDF), and low (LDF) density fractions. Morphological and flow cytometrical observations demonstrated that HDF, IDF, and LDF were basal, spinous, and granular cell-rich fractions, respectively. Membrane fluidity of the fractionated keratinocytes was measured by diphenylhexatriene fluorescence polarization. Polarization (p)-value of keratinocytes was negatively correlated with temperature. At each temperature, HDF cells showed a lower p-value than IDF or HDF cells except at 40° C. Since a low p-value indicates a high degree of Brownian motion, membrane fluidity is higher in basal cells and lower in spinous and granular cells. Our results indicate that membrane fluidity of guinea-pig keratinocytes decreases during their maturation.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

References

  • Abe J, Nomura H, Hachisuka H, Mori O, Sasai Y (1985) Electron microscopic observation of epidermal cells separated by colloidal silica density gradient centrifugation. Jpn J Dermatol 95:807–809

    Google Scholar 

  • Barton SP, Marks R (1981) Changes in suspensions of human keratinocytes due to trypsin. Arch Dermatol Res 271:245–257

    Google Scholar 

  • Cercek L, Cercek B, Ockey CH (1973) Structuredness of the cytoplasmic matrix and Michaelis-Menten constants for the hydrolysis of FDA during the cell cycle in Chinese hamster ovary cells. Biophysik 10:187–194

    Google Scholar 

  • Goldenhersch MA, Good RA, Sarker NH, Safai B (1982) Separation of epidermal cells by density gradient centrifugation on a continuous colloidal silica (Percoll) gradient. Anal Biochem 119:246–252

    Google Scholar 

  • Gray GM, King IA, Yardley HJ (1980) The plasma membrane of Malpighian cells from pig epidermis: isolation and lipid and protein composition. Br J Dermatol 103:505–515

    Google Scholar 

  • Hachisuka H, Nomura H, Sasai Y, Shiotsuki K, Yokoyama MM (1986) Change of intracellular fluidity during keratinocyte differentiation measured by fluorescence polarization. Cell Tissue Res 246:557–560

    Google Scholar 

  • Jahnig F (1979) Structural order of lipids and proteins in membranes: Evaluation of fluorescence anisotropy data. Proc Natl Acad Sci USA 76:6361–6365

    Google Scholar 

  • Kaplan J, Canonico PG, Caspary WJ (1973) Electron spin resonance studies of spin-labeled mammalian cells by detection of surface-membrane signals. Proc Natl Acad Sci USA 70:66–70

    Google Scholar 

  • Lampe MA, Williams ML, Elias PM (1983) Human epidermal lipids: characterization and modulations during differentiation. J Lipid Res 24:131–140

    Google Scholar 

  • Lentz BR, Barenholz Y, Thompson TE (1976) Fluorescence depolarization studies of phase transitions and fluidity in phospholipid bilayers. 2. Two-component phosphatidylcholine liposomes. Biochemistry 15:4529–4537

    Google Scholar 

  • Long VJW (1970) Variations in lipid composition at different dephts in the cow snout epidermis. J Invest Dermatol 55:269–273

    Google Scholar 

  • Nomura H, Hachisuka H, Mori O, Sakamoto F, Sasai Y (1985) Cytofluorometric study of thiol and disulphide groups in guinea pig epidermis. Acta Histochem 76:213–218

    Google Scholar 

  • Parasassi T, Conti F, Glaser M, Gratton E (1984) Detection of phospholipid phase separation: A multifrequency phase fluorimetry study of 1,6-diphenyl-1,3,5-hexatriene fluorescence. J Biol Chem 259:14011–14017

    Google Scholar 

  • Rotman B, Papermaster BW (1966) Membrane properties of living mammalian cells as studies by enzymatic hydrolysis of fluorogenic esters. Proc Natl Acad Sci USA 55:134–141

    Google Scholar 

  • Rowden G (1975) Ultrastructural studies of keratinized epithelia of the mouse. III. Determination of the volumes of nuclei and cytoplasm of cells in murine epidermis. J Invest Dermatol 64:1–3

    Google Scholar 

  • Salzman GC, Mullaney PF, Price BJ (1979) Light-scattering approaches to cell characterization. In: Melamed MR, Mullaney PF, Mendelsohn HL (eds) Flow cytometry and sorting. Wiley Medical Publication, New York, pp 105–124

    Google Scholar 

  • Sasai Y, Hachisuka H, Mori O, Nomura H (1984) Separation of keratinocytes by density gradient centrifugation for DNA cytofluorometry. Histochemistry 80:133–136

    Google Scholar 

  • Shinitzky M, Inbar M (1974) Difference in microviscosity induced by different cholesterol levels in the surface membrane lipid layer of normal lymphocytes and malignant lymphoma cells. J Mol Biol 85:603–615

    Google Scholar 

  • Singer SJ, Nicolson GL (1972) The fluid mosaic model of the structure of cell membranes: Cell membranes are viewed as two-dimensional solutions of oriented globular proteins and lipids. Science 175:720–731

    Google Scholar 

  • Sun T-T, Green H (1976) Differentiation of the epidermal keratinocyte in culture: Formation of the cornified envelope. Cell 9:511–521

    Google Scholar 

  • Tanaka T, Hidaka T, Ogura R, Sugiyama M (1988) Change of membrane fluidity of Na+, K+-ATPase activity during cellular differentiation in the guinea pig epidermis. Arch Dermatol Res 280:29–32

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hachisuka, H., Nomura, H., Mori, O. et al. Alterations in membrane fluidity during keratinocyte differentiation measured by fluorescence polarization. Cell Tissue Res 260, 207–210 (1990). https://doi.org/10.1007/BF00297507

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00297507

Key words

Navigation