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Biophys J. Nov 1993; 65(5): 2021–2040.
PMCID: PMC1225938

Confined lateral diffusion of membrane receptors as studied by single particle tracking (nanovid microscopy). Effects of calcium-induced differentiation in cultured epithelial cells.


The movements of E-cadherin, epidermal growth factor receptor, and transferrin receptor in the plasma membrane of a cultured mouse keratinocyte cell line were studied using both single particle tracking (SPT; nanovid microscopy) and fluorescence photobleaching recovery (FPR). In the SPT technique, the receptor molecules are labeled with 40 nm-phi colloidal gold particles, and their movements are followed by video-enhanced differential interference contrast microscopy at a temporal resolution of 33 ms and at a nanometer-level spatial precision. The trajectories of the receptor molecules obtained by SPT were analyzed by developing a method that is based on the plot of the mean-square displacement against time. Four characteristic types of motion were observed: (a) stationary mode, in which the microscopic diffusion coefficient is less than 4.6 x 10(-12) cm2/s; (b) simple Brownian diffusion mode; (c) directed diffusion mode, in which unidirectional movements are superimposed on random motion; and (d) confined diffusion mode, in which particles undergoing Brownian diffusion (microscopic diffusion coefficient between 4.6 x 10(-12) and 1 x 10(-9) cm2/s) are confined within a limited area, probably by the membrane-associated cytoskeleton network. Comparison of these data obtained by SPT with those obtained by FPR suggests that the plasma membrane is compartmentalized into many small domains 300-600 nm in diameter (0.04-0.24 microns2 in area), in which receptor molecules are confined in the time scale of 3-30 s, and that the long-range diffusion observed by FPR can occur by successive movements of the receptors to adjacent compartments. Calcium-induced differentiation decreases the sum of the percentages of molecules in the directed diffusion and the stationary modes outside of the cell-cell contact regions on the cell surface (which is proposed to be the percentage of E-cadherin bound to the cytoskeleton/membrane-skeleton), from approximately 60% to 8% (low- and high-calcium mediums, respectively).

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  • Abney JR, Scalettar BA, Owicki JC. Self diffusion of interacting membrane proteins. Biophys J. 1989 May;55(5):817–833. [PMC free article] [PubMed]
  • Anderson CM, Georgiou GN, Morrison IE, Stevenson GV, Cherry RJ. Tracking of cell surface receptors by fluorescence digital imaging microscopy using a charge-coupled device camera. Low-density lipoprotein and influenza virus receptor mobility at 4 degrees C. J Cell Sci. 1992 Feb;101(Pt 2):415–425. [PubMed]
  • Austin RH, Chan SS, Jovin TM. Rotational diffusion of cell surface components by time-resolved phosphorescence anisotropy. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5650–5654. [PMC free article] [PubMed]
  • Axelrod D, Koppel DE, Schlessinger J, Elson E, Webb WW. Mobility measurement by analysis of fluorescence photobleaching recovery kinetics. Biophys J. 1976 Sep;16(9):1055–1069. [PMC free article] [PubMed]
  • Bennett V. Spectrin-based membrane skeleton: a multipotential adaptor between plasma membrane and cytoplasm. Physiol Rev. 1990 Oct;70(4):1029–1065. [PubMed]
  • Chang CH, Takeuchi H, Ito T, Machida K, Ohnishi S. Lateral mobility of erythrocyte membrane proteins studied by the fluorescence photobleaching recovery technique. J Biochem. 1981 Oct;90(4):997–1004. [PubMed]
  • Cherry RJ, Bürkli A, Busslinger M, Schneider G, Parish GR. Rotational diffusion of band 3 proteins in the human erythrocyte membrane. Nature. 1976 Sep 30;263(5576):389–393. [PubMed]
  • Cherry RJ. Rotational and lateral diffusion of membrane proteins. Biochim Biophys Acta. 1979 Dec 20;559(4):289–327. [PubMed]
  • Cherry RJ. Keeping track of cell surface receptor. Trends Cell Biol. 1992 Aug;2(8):242–244. [PubMed]
  • De Brabander M, Geuens G, Nuydens R, Moeremans M, De Mey J. Probing microtubule-dependent intracellular motility with nanometre particle video ultramicroscopy (nanovid ultramicroscopy). Cytobios. 1985;43(174S):273–283. [PubMed]
  • De Brabander M, Nuydens R, Geuens G, Moeremans M, De Mey J. The use of submicroscopic gold particles combined with video contrast enhancement as a simple molecular probe for the living cell. Cell Motil Cytoskeleton. 1986;6(2):105–113. [PubMed]
  • De Brabander M, Nuydens R, Geerts H, Hopkins CR. Dynamic behavior of the transferrin receptor followed in living epidermoid carcinoma (A431) cells with nanovid microscopy. Cell Motil Cytoskeleton. 1988;9(1):30–47. [PubMed]
  • de Brabander M, Nuydens R, Ishihara A, Holifield B, Jacobson K, Geerts H. Lateral diffusion and retrograde movements of individual cell surface components on single motile cells observed with Nanovid microscopy. J Cell Biol. 1991 Jan;112(1):111–124. [PMC free article] [PubMed]
  • den Hartigh JC, van Bergen en Henegouwen PM, Verkleij AJ, Boonstra J. The EGF receptor is an actin-binding protein. J Cell Biol. 1992 Oct;119(2):349–355. [PMC free article] [PubMed]
  • Dubinsky JM, Loftus DJ, Fischbach GD, Elson EL. Formation of acetylcholine receptor clusters in chick myotubes: migration or new insertion? J Cell Biol. 1989 Oct;109(4 Pt 1):1733–1743. [PMC free article] [PubMed]
  • Edidin M, Stroynowski I. Differences between the lateral organization of conventional and inositol phospholipid-anchored membrane proteins. A further definition of micrometer scale membrane domains. J Cell Biol. 1991 Mar;112(6):1143–1150. [PMC free article] [PubMed]
  • Edidin M, Kuo SC, Sheetz MP. Lateral movements of membrane glycoproteins restricted by dynamic cytoplasmic barriers. Science. 1991 Nov 29;254(5036):1379–1382. [PubMed]
  • Geerts H, De Brabander M, Nuydens R, Geuens S, Moeremans M, De Mey J, Hollenbeck P. Nanovid tracking: a new automatic method for the study of mobility in living cells based on colloidal gold and video microscopy. Biophys J. 1987 Nov;52(5):775–782. [PMC free article] [PubMed]
  • Geerts H, de Brabander M, Nuydens R. Nanovid microscopy. Nature. 1991 Jun 27;351(6329):765–766. [PubMed]
  • Gelles J, Schnapp BJ, Sheetz MP. Tracking kinesin-driven movements with nanometre-scale precision. Nature. 1988 Feb 4;331(6155):450–453. [PubMed]
  • Goldstein JL, Brown MS, Anderson RG, Russell DW, Schneider WJ. Receptor-mediated endocytosis: concepts emerging from the LDL receptor system. Annu Rev Cell Biol. 1985;1:1–39. [PubMed]
  • Hennings H, Michael D, Cheng C, Steinert P, Holbrook K, Yuspa SH. Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell. 1980 Jan;19(1):245–254. [PubMed]
  • Hennings H, Kruszewski FH, Yuspa SH, Tucker RW. Intracellular calcium alterations in response to increased external calcium in normal and neoplastic keratinocytes. Carcinogenesis. 1989 Apr;10(4):777–780. [PubMed]
  • Hillman GM, Schlessinger J. Lateral diffusion of epidermal growth factor complexed to its surface receptors does not account for the thermal sensitivity of patch formation and endocytosis. Biochemistry. 1982 Mar 30;21(7):1667–1672. [PubMed]
  • Hirano S, Nose A, Hatta K, Kawakami A, Takeichi M. Calcium-dependent cell-cell adhesion molecules (cadherins): subclass specificities and possible involvement of actin bundles. J Cell Biol. 1987 Dec;105(6 Pt 1):2501–2510. [PMC free article] [PubMed]
  • Iacopetta BJ, Rothenberger S, Kühn LC. A role for the cytoplasmic domain in transferrin receptor sorting and coated pit formation during endocytosis. Cell. 1988 Aug 12;54(4):485–489. [PubMed]
  • Jacobson K, Ishihara A, Inman R. Lateral diffusion of proteins in membranes. Annu Rev Physiol. 1987;49:163–175. [PubMed]
  • Johnson P, Garland PB. Depolarization of fluorescence depletion. A microscopic method for measuring rotational diffusion of membrane proteins on the surface of a single cell. FEBS Lett. 1981 Sep 28;132(2):252–256. [PubMed]
  • Kemler R, Ozawa M. Uvomorulin-catenin complex: cytoplasmic anchorage of a Ca2+-dependent cell adhesion molecule. Bioessays. 1989 Oct;11(4):88–91. [PubMed]
  • Kucik DF, Elson EL, Sheetz MP. Forward transport of glycoproteins on leading lamellipodia in locomoting cells. Nature. 1989 Jul 27;340(6231):315–317. [PubMed]
  • Kulesz-Martin M, Kilkenny AE, Holbrook KA, Digernes V, Yuspa SH. Properties of carcinogen altered mouse epidermal cells resistant to calcium-induced terminal differentiation. Carcinogenesis. 1983 Nov;4(11):1367–1377. [PubMed]
  • Kusumi A, Sakaki T, Yoshizawa T, Ohnishi S. Protein-lipid interaction in rhodopsin recombinant membranes as studied by protein rotational mobility and lipid alkyl chain flexibility measurements. J Biochem. 1980 Oct;88(4):1103–1111. [PubMed]
  • Kusumi A, Hyde JS. Spin-label saturation-transfer electron spin resonance detection of transient association of rhodopsin in reconstituted membranes. Biochemistry. 1982 Nov 9;21(23):5978–5983. [PubMed]
  • Kusumi A, Subczynski WK, Pasenkiewicz-Gierula M, Hyde JS, Merkle H. Spin-label studies on phosphatidylcholine-cholesterol membranes: effects of alkyl chain length and unsaturation in the fluid phase. Biochim Biophys Acta. 1986 Jan 29;854(2):307–317. [PubMed]
  • Landreth GE, Williams LK, Rieser GD. Association of the epidermal growth factor receptor kinase with the detergent-insoluble cytoskeleton of A431 cells. J Cell Biol. 1985 Oct;101(4):1341–1350. [PMC free article] [PubMed]
  • Lee GM, Ishihara A, Jacobson KA. Direct observation of brownian motion of lipids in a membrane. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6274–6278. [PMC free article] [PubMed]
  • Livneh E, Benveniste M, Prywes R, Felder S, Kam Z, Schlessinger J. Large deletions in the cytoplasmic kinase domain of the epidermal growth factor receptor do not affect its laternal mobility. J Cell Biol. 1986 Aug;103(2):327–331. [PMC free article] [PubMed]
  • McCrea PD, Gumbiner BM. Purification of a 92-kDa cytoplasmic protein tightly associated with the cell-cell adhesion molecule E-cadherin (uvomorulin). Characterization and extractability of the protein complex from the cell cytostructure. J Biol Chem. 1991 Mar 5;266(7):4514–4520. [PubMed]
  • McNeill H, Ozawa M, Kemler R, Nelson WJ. Novel function of the cell adhesion molecule uvomorulin as an inducer of cell surface polarity. Cell. 1990 Jul 27;62(2):309–316. [PubMed]
  • Mecham RP, Whitehouse L, Hay M, Hinek A, Sheetz MP. Ligand affinity of the 67-kD elastin/laminin binding protein is modulated by the protein's lectin domain: visualization of elastin/laminin-receptor complexes with gold-tagged ligands. J Cell Biol. 1991 Apr;113(1):187–194. [PMC free article] [PubMed]
  • Metzger H. Transmembrane signaling: the joy of aggregation. J Immunol. 1992 Sep 1;149(5):1477–1487. [PubMed]
  • Miller K, Shipman M, Trowbridge IS, Hopkins CR. Transferrin receptors promote the formation of clathrin lattices. Cell. 1991 May 17;65(4):621–632. [PubMed]
  • Moore C, Boxer D, Garland P. Phosphorescence depolarization and the measurement of rotational motion of proteins in membranes. FEBS Lett. 1979 Dec 1;108(1):161–166. [PubMed]
  • Nagafuchi A, Takeichi M. Cell binding function of E-cadherin is regulated by the cytoplasmic domain. EMBO J. 1988 Dec 1;7(12):3679–3684. [PMC free article] [PubMed]
  • Nagafuchi A, Shirayoshi Y, Okazaki K, Yasuda K, Takeichi M. Transformation of cell adhesion properties by exogenously introduced E-cadherin cDNA. Nature. 1987 Sep 24;329(6137):341–343. [PubMed]
  • Ozawa M, Baribault H, Kemler R. The cytoplasmic domain of the cell adhesion molecule uvomorulin associates with three independent proteins structurally related in different species. EMBO J. 1989 Jun;8(6):1711–1717. [PMC free article] [PubMed]
  • Ozawa M, Ringwald M, Kemler R. Uvomorulin-catenin complex formation is regulated by a specific domain in the cytoplasmic region of the cell adhesion molecule. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4246–4250. [PMC free article] [PubMed]
  • Peters R, Cherry RJ. Lateral and rotational diffusion of bacteriorhodopsin in lipid bilayers: experimental test of the Saffman-Delbrück equations. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4317–4321. [PMC free article] [PubMed]
  • Qian H, Sheetz MP, Elson EL. Single particle tracking. Analysis of diffusion and flow in two-dimensional systems. Biophys J. 1991 Oct;60(4):910–921. [PMC free article] [PubMed]
  • Ryan TA, Myers J, Holowka D, Baird B, Webb WW. Molecular crowding on the cell surface. Science. 1988 Jan 1;239(4835):61–64. [PubMed]
  • Saffman PG, Delbrück M. Brownian motion in biological membranes. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3111–3113. [PMC free article] [PubMed]
  • Saxton MJ. Lateral diffusion in an archipelago. Effects of impermeable patches on diffusion in a cell membrane. Biophys J. 1982 Aug;39(2):165–173. [PMC free article] [PubMed]
  • Saxton MJ. Lateral diffusion in an archipelago. The effect of mobile obstacles. Biophys J. 1987 Dec;52(6):989–997. [PMC free article] [PubMed]
  • Saxton MJ. Lateral diffusion in an archipelago. Distance dependence of the diffusion coefficient. Biophys J. 1989 Sep;56(3):615–622. [PMC free article] [PubMed]
  • Saxton MJ. The spectrin network as a barrier to lateral diffusion in erythrocytes. A percolation analysis. Biophys J. 1989 Jan;55(1):21–28. [PMC free article] [PubMed]
  • Saxton MJ. The membrane skeleton of erythrocytes. A percolation model. Biophys J. 1990 Jun;57(6):1167–1177. [PMC free article] [PubMed]
  • Saxton MJ. The membrane skeleton of erythrocytes: models of its effect on lateral diffusion. Int J Biochem. 1990;22(8):801–809. [PubMed]
  • Saxton MJ. Lateral diffusion and aggregation. A Monte Carlo study. Biophys J. 1992 Jan;61(1):119–128. [PMC free article] [PubMed]
  • Saxton MJ. Lateral diffusion in an archipelago. Single-particle diffusion. Biophys J. 1993 Jun;64(6):1766–1780. [PMC free article] [PubMed]
  • Schlessinger J. Allosteric regulation of the epidermal growth factor receptor kinase. J Cell Biol. 1986 Dec;103(6 Pt 1):2067–2072. [PMC free article] [PubMed]
  • Schlessinger J, Schreiber AB, Levi A, Lax I, Libermann T, Yarden Y. Regulation of cell proliferation by epidermal growth factor. CRC Crit Rev Biochem. 1983;14(2):93–111. [PubMed]
  • Schnapp BJ, Gelles J, Sheetz MP. Nanometer-scale measurements using video light microscopy. Cell Motil Cytoskeleton. 1988;10(1-2):47–53. [PubMed]
  • Sheetz MP. Glycoprotein motility and dynamic domains in fluid plasma membranes. Annu Rev Biophys Biomol Struct. 1993;22:417–431. [PubMed]
  • Sheetz MP, Schindler M, Koppel DE. Lateral mobility of integral membrane proteins is increased in spherocytic erythrocytes. Nature. 1980 Jun 12;285(5765):510–511. [PubMed]
  • Sheetz MP, Turney S, Qian H, Elson EL. Nanometre-level analysis demonstrates that lipid flow does not drive membrane glycoprotein movements. Nature. 1989 Jul 27;340(6231):284–288. [PubMed]
  • Sheetz MP, Baumrind NL, Wayne DB, Pearlman AL. Concentration of membrane antigens by forward transport and trapping in neuronal growth cones. Cell. 1990 Apr 20;61(2):231–241. [PubMed]
  • Takeichi M. The cadherins: cell-cell adhesion molecules controlling animal morphogenesis. Development. 1988 Apr;102(4):639–655. [PubMed]
  • Tsuji A, Kawasaki K, Ohnishi S, Merkle H, Kusumi A. Regulation of band 3 mobilities in erythrocyte ghost membranes by protein association and cytoskeletal meshwork. Biochemistry. 1988 Sep 20;27(19):7447–7452. [PubMed]
  • Tsuji A, Ohnishi S. Restriction of the lateral motion of band 3 in the erythrocyte membrane by the cytoskeletal network: dependence on spectrin association state. Biochemistry. 1986 Oct 7;25(20):6133–6139. [PubMed]
  • van Beijeren H, Kutner R. Mean square displacement of a tracer particle in a hard-core lattice gas. Phys Rev Lett. 1985 Jul 8;55(2):238–241. [PubMed]
  • Vaz WL, Melo EC, Thompson TE. Translational diffusion and fluid domain connectivity in a two-component, two-phase phospholipid bilayer. Biophys J. 1989 Nov;56(5):869–876. [PMC free article] [PubMed]
  • Wiegant FA, Blok FJ, Defize LH, Linnemans WA, Verkley AJ, Boonstra J. Epidermal growth factor receptors associated to cytoskeletal elements of epidermoid carcinoma (A431) cells. J Cell Biol. 1986 Jul;103(1):87–94. [PMC free article] [PubMed]
  • Yoshitake S, Imagawa M, Ishikawa E, Niitsu Y, Urushizaki I, Nishiura M, Kanazawa R, Kurosaki H, Tachibana S, Nakazawa N, et al. Mild and efficient conjugation of rabbit Fab' and horseradish peroxidase using a maleimide compound and its use for enzyme immunoassay. J Biochem. 1982 Nov;92(5):1413–1424. [PubMed]
  • Yuspa SH, Morgan DL. Mouse skin cells resistant to terminal differentiation associated with initiation of carcinogenesis. Nature. 1981 Sep 3;293(5827):72–74. [PubMed]
  • Zhang F, Crise B, Su B, Hou Y, Rose JK, Bothwell A, Jacobson K. Lateral diffusion of membrane-spanning and glycosylphosphatidylinositol-linked proteins: toward establishing rules governing the lateral mobility of membrane proteins. J Cell Biol. 1991 Oct;115(1):75–84. [PMC free article] [PubMed]
  • Zidovetzki R, Bartholdi M, Arndt-Jovin D, Jovin TM. Rotational dynamics of the Fc receptor for immunoglobulin E on histamine-releasing rat basophilic leukemia cells. Biochemistry. 1986 Jul 29;25(15):4397–4401. [PubMed]

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