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Single-membrane and cell-to-cell permeability properties of dissociated embryonic chick lens cells

The Journal of Membrane Biology volume 128, pages 91–102 (1992)Cite this article

Summary

Ion channels are believed to play an important role in the maintenance of lens transparency. In order to ascribe junctional and nonjunctional permeability properties to specific lens cell types, embryonic chick lenses were enzymatically dissociated into cell clusters, cell pairs and single cells, and both cell-to-cell and single-membrane permeability properties were characterized with the patch-clamp technique. Double patch-clamp experiments and single patch-clamp experiments with Lucifer yellow in the pipette demonstrated that the cells in the dissociated preparation were well coupled, the average conductance between pairs being 42 ± 27 nS. Double patch-clamp experiments also revealed single cell-to-cell channel events with a predominant unitary conductance of 286 ± 38 pS. Whole-cell measurements of surface membrane conductance indicate heterogeneity within the population of dissociated embryonic chick lens cells: 63% of the cells have a voltage-independent leak current, 14% of the cells have a potassium-selective inward rectifier current, and 23% of the cells have a current which turns off with positive voltage on a time scale on the order of seconds. The time constant for this turnoff is voltage dependent.

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References

  • Atkinson, M.M., Sheridan, J.D. 1988. Altered junctional permeability between cells transformed by v-ras, v-mos, or v-src. Am. J. Physiol. 255:C674-C683

    Google Scholar 

  • Bunce, G.E., Kinoshita, J.H., Horwitz, J. 1990. Nutritional factors in cataract. Annu. Rev. Nutr. 10:233–254

    Google Scholar 

  • Cooper, K, Gates, P., Rae, J.L., Dewey, J. 1990. Electrophysiology of cultured human lens epithelial cells. J. Membrane Biol. 117:285–298

    Google Scholar 

  • Cooper, K.E., Rae, J.L., Dewey, J. 1991. Inwardly rectifying potassium current in mammalian lens epithelial cells. Am. J. Physiol. 61:C115-C123

    Google Scholar 

  • Cooper, K., Rae, J.L., Gates, P. 1989. Membrane and junctional properties of dissociated frog lens epithelial cells. J. Membrane Biol. 111:215–227

    Google Scholar 

  • Cooper, K.E., Tang, J.M., Rae, J.L., Eisenberg, R.S. 1986. A cation channel in frog lens epithelia responsive to pressure and calcium. J. Membrane Biol. 93:259–269

    Google Scholar 

  • De Hemptinne, A. Marrannes, R. Vanheel, B. 1983. Influence of organic acids on intracellular pH. Am. J. Physiol. 245:C178-C183

    Google Scholar 

  • Duncan, G., Bushell, A.R. 1975. Ion analyses of human cataractous lenses. Exp. Eye Res. 20:223–230

    Google Scholar 

  • Duncan, G., Bushell, A.R. 1979. Relationships between colour, sodium and protein content in individual senile human cataractous lenses. Ophthalmic Res. 11:397–404

    Google Scholar 

  • Duncan, G., Jacob, T.J.C. 1984. Calcium and the physiology of cataract. In: Human Cataract Formation. J. Nugent and J. Whelan, editor, pp. 132–152. Pitman, Bath, UK

    Google Scholar 

  • Duncan, G., Stewart, S., Prescott, A.R., Warn, R.M. 1988. Membrane and junctional properties of the isolated frog lens epithelium. J. Membrane Biol. 102:195–204

    Google Scholar 

  • Ehring, G.R., Zampighi, G.A., Horwitz, J., Bok, D., Hall, J.E. 1990. Properties of channels reconstituted from the major intrinsic protein of lens fiber membranes. J. Gen. Physiol. 96:631–664

    Google Scholar 

  • Goodenough, D.A., Dick, J.S.B., Lyons, J.E. 1980. Lens metabolic cooperation: A study of mouse lens permeability visualized with freeze-substitution autoradiography and electron microscopy. J. Cell Biol. 86:576–589

    Google Scholar 

  • Jacob, T.J.C. 1986. The electrophysiology of cultured human lens epithelial cells. J. Physiol. 377:39P

  • Jacob, T.J.C., Bangham, J.A., Duncan, G. 1985. Characterization of a cation channel on the apical surface of the frog lens epithelium. Q. J. Exp. Physiol. 70:403–421

    Google Scholar 

  • Lo, W.K., Harding, C.V. 1986. Structure and distribution of gap junctions in lens epithelium and fiber cells. Cell Tissue Res. 214:253–263

    Google Scholar 

  • Mathias, R.T., Rae, J.L. 1985a. Steady state voltages in the frog lens. Curr. Eye Res. 4:421–430

    Google Scholar 

  • Mathias R.T. Rae, J.L. 1985b. Transport properties of the lens. Am. J. Physiol. 249:C181-C190

    Google Scholar 

  • Mathias, R.T., Rae, J.L., Ebihara, L., McCarthy, R.T. 1985. The localization of transport properties in the frog lens. Biophys. J. 48:423–434

    Google Scholar 

  • Menko, A.S., Klukas, K.A., Liu, T.F., Quade, B., Sas, D.F., Preus, D.M., Johnson, R.G. 1987. Junctions between lens cells in differentiating cultures: Structure, formation, intercellular permeability, and junctional protein expression. Dev. Biol. 123:307–320

    Google Scholar 

  • Miller, A.G., Hall, J.E., Zampighi, G.A. 1990. Cell-to-cell permeability in dissociated embryonic chicken lenses. Biophys. J. 57:245a

  • Miller, T.M., Goodenough, D.A. 1986. Evidence for two physiologically distinct gap junctions expressed by the chick lens epithelial cell. J. Cell Biol. 102:194–199

    Google Scholar 

  • Neyton, J., Trautmann, A. 1985. Single-channel currents of an intercellular junction. Nature 317:331–335

    Google Scholar 

  • Patmore, L., Duncan, G. 1981. The physiology of lens membranes. In: Mechanisms of Cataract Formation in the Human Lens. G. Duncan, editor, pp. 193–218. Academic, San Francisco

    Google Scholar 

  • Rae, J.L. 1985. The application of patch clamp methods to ocular epithelia. Curr. Eye Res. 4:409–420

    Google Scholar 

  • Rae, J.L. 1986. Potassium channels from chick lens epithelium. Fed. Proc. 45:2718–2722

    Google Scholar 

  • Rae, J.L., Cooper, K.E. 1989. Potassium channels in chick lens epithelium change with maturation. Lens Eye Toxicol. Res. 6:833–843

    Google Scholar 

  • Rae, J.L., Dewey, J., Rae, J.S., Cooper, K.E. 1990. A maxi calcium activated potassium channel from chick lens epithelium. Curr. Eye Res. 9:847–861

    Google Scholar 

  • Rae, J.L., Levis, R.A. 1984. Patch voltage clamp of lens epithelial cells: Theory and practice. Mol. Physiol. 6:115–162

    Google Scholar 

  • Robinson, K.R., Patterson, J.W. 1983. Localization of steady currents in the lens. Curr. Eye Res. 2:843–847

    Google Scholar 

  • Scheutze, S.M., Goodenough, D.A. 1982. Dye transfer between cells of the embryonic chick lens becomes less sensitive to CO2 with development. J. Cell Biol. 92:694–705

    Google Scholar 

  • Shinohara, T., Piatigorsky, J. 1977. Regulation of protein synthesis, intracellular electrolytes and cataract formation in vitro. Nature 270:406–411

    Google Scholar 

  • Stewart, S., Duncan, G., Marcantonio, J.M., Precott, A.R. 1988. Membrane and communication properties of cultured human lens epithelial cells. Invest. Ophthalmol. Vis. Sci. 29:1713–1725

    Google Scholar 

  • Unakar, N.J., Tsui, J.Y. 1980. Sodium-potassium-dependent ATPase 1. Cytochemical localization in normal and cataractous rat lenses. Invest. Ophthalmol. Vis. Sci. 19:630–641

    Google Scholar 

  • Wind, B.E., Walsh, S., Patterson, J.W. 1988. Equatorial potassium currents in lenses. Exp. Eye Res. 46:117–130

    Google Scholar 

  • Zampighi, G.A., Hall, J.E., Ehring, G.R., Simon, S.A. 1989. The structural organization and protein composition of lens fiber junctions. J. Cell Biol. 108:2255–2276

    Google Scholar 

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Affiliations

  1. Department of Physiology and Biophysics, University of California at Irvine, 92715, Irvine, California

    A. G. Millert & J. E. Hall

  2. Department of Cell Biology and Anatomy, University of California at Los Angeles, 90024, Los Angeles, California

    G. A. Zampighi

Authors
  1. A. G. Millert
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  2. G. A. Zampighi
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  3. J. E. Hall
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Additional information

The authors would like to thank Mary Hawley for her technical support and for the preparation of Fig. 1, Richard Wetts for his help with the nuclear staining, Mike Kreman for his help with Fig. 3 and Ruth Davis for helpful comments on this manuscript. This work was supported by NIH grants EY-06884 to JEH and EY-04110 to GAZ.

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Millert, A.G., Zampighi, G.A. & Hall, J.E. Single-membrane and cell-to-cell permeability properties of dissociated embryonic chick lens cells. J. Membarin Biol. 128, 91–102 (1992). https://doi.org/10.1007/BF00231882

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  • DOI: https://doi.org/10.1007/BF00231882

Key words

  • lens
  • embryonic
  • gap junctions
  • ion channels