Summary
Mammalian cells in culture were exposed to cyclic AMP, dibutyrul cyclic AMP, the phosphodiesterase inhibitor caffeine, or a combination of the last two, while junctional molecular transfer was probed with the series of microinjected, fluorescentlabelled linear molecules Glu, Glu-Glu, Glu-Glu-Glu, and Leu-Leu-Leu-Glu-Glu. The junctional permeability for these molecules increased with each of the agents, most markedly with the dibutyryl cyclic AMP-caffeine combination, as the intracellular cyclic nucleotide concentration rose. The junctional permeability effect developed over several hours. When probed with molecules close to the limit of cell-to-cell channel permeation (the most sensitive setting), the effect was detectable both, as an increase in the (relative) junctional transit rate and as an increase in the number of transferring cell interfaces in the test populations. The number of transferring cell interfaces reached a maximum by 4 hr, when the junctional transit rate, hence the junctional permeability, was still rising. Nonjunctional membrane permeability for the probe molecules, as determined by intracellular fluorescence loss, was not significantly changed (nor was there significant nonjunctional cell-to-cell transfer of molecules before or after the treatments). The rise in junctional permeability was associated with an increase in the number of gap junctional membrane particles, as determined by freeze-fracture electron microscopy: the average size of the particle clusters increased, and the frequency of the clusters increased, particularly that of the smaller (and presumably newer) clusters. This effect was blocked by treatments with the protein synthesis inhibitors cycloheximide or puromycin. These agents caused particle diminution (diminution of cluster frequency but not of average cluster size), with or without cyclic nucleotide. The junctional effects may represent a cyclic AMP-promoted proliferation of cell-to-cell channels. Some physiological implications, in particular, implications for hormone-regulated tissues, are discussed.
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Azarnia, R., Larsen, W. J., Loewenstein, W. R. 1974. The membrane junctions in communicating and non-communicating cells, their hybrids and segregants.Proc. Natl. Acad. Sci. USA 71:880–884
Bean, C. P. 1972. The physics of porous membranes.In: Membranes. Vol. 1, Macroscopic Systems and Models. G. Eisenman, editor. Marcel Decker, New York
Blalock, J. E., Stanton, J. D. 1980. Common pathways of interferon and hormonal action.Nature (London) 283:406–408
Boone, C. W. 1975. Malignant hemangioendotheliomas produced by subcutaneous inoculation of Balb/3T3 cells attached to glass beads.Science 188:68–70
Caspar, D. L. C., Goodenough, D. A., Makowski, L., Phillips, W. C. 1977. Gap junction structures. I. Correlated electron microscopy and X-ray diffraction.J. Cell Biol. 74:605–628
Caveney, S. 1978. Intercellular communication in insect development is hormonally controlled.Science 199:192–195
Caveney, S., Blennerhassett, M. G. 1980. Elevation of ionic conductance between insect epidermal cells by β-ecdysonein vitro.J. Insect Physiol. 26:13–25
Dahl, G., Azarnia, R., Werner, R. 1981. Induction of cell-cell channel formation by mRNA.Nature (London) 289:683–685
Dahl, G., Berger, W. 1978. Nexus formation in the myometrium during parturition and induced by estrogen.Cell Biol. Int. Rep. 2:381–387
Decker, R. J. 1976. Hormonal regulation of gap junction differentiation.J. Cell Biol. 69:669–685
Dedman, J. R., Brinkley, B. R., Means, A. R. 1979. Regulation of microfilaments and microtubules by calcium and cyclic AMP.Adv. Cyclic Nucleotide Res. 11:131–174
Epstein, M. L., Sheridan, J. D., Johnson, R. G. 1977. Formation of low resistance junctionsin vitro in the absence of protein synthesis and ATP production.Exp. Cell Res. 104:25–30
Flagg-Newton, J. L. 1979. Cyclic AMP increases junctional permeability in cultured mammalian cells.Biophys. J. 25:80a
Flagg-Newton, J. L. 1980. The permeability of the cell-to-cell membrane channel and its regulation in mammalian cell junction.In Vitro 16(12):1043–1048
FLagg-Newton, J. L., Loewenstein, W. R. 1979. Experimental depression of junctional membrane permeability in mammalian cell culture. A study with tracer molecules in the 300 to 800 dalton range.J. Membrane Biol. 50:65–100
Flagg-Newton, J. L., Loewenstein, W. R., 1980. Asymmetrically permeable membrane channels in cell junction.Science 207:771–773
Flagg-Newton, J. L., Loewenstein, W. R. 1981. Cell junction and cyclic AMP. II. Modulation of junctional membrane permeability, dependent on serum and cell density.J. Membrane Biol. 63:123–131
Flagg-Newton, J. L., Simpson, I., Loewenstein, W. R. 1979. Permeability of the cell-to-cell membrane channels in mammalian cell junction.Science 205:404–407
Furshpan, E. J., Potter, D. D. 1968. Low resistance junctions between cells in embryo and tissue culture.Curr. Topics Dev. Biol. 3:95–127
Garfield, R. E., Merrett, D., Grover, A. K. 1980. Gap junction formation and regulation in myometrium.Am J. Physiol. Cell Physiol. 8:C217-C228
Gilula, N. B., Reeves, O. R., Steinbach, A. 1972. Metabolic coupling, ionic coupling and cell contacts.Nature (London) 235:262–265
Goodenough, D. A., Revel, J.-P. 1970. A fine structural analysis of intercellular junctions in the mouse liver.J. Cell Biol. 45:272–284
Hax, W. M. A., Venrooij, G. E. P. M. van, Vossenberg, J. B. J. 1974. Cell communication: A cyclic-AMP mediated phenomenon.J. Membrane Biol. 19:253–266
Ito, S., Sato, E., Loewenstein, W. R. 1974. Studies on the formation of a permeable cell membrane junction. II. Evolving junctional conductance and junctional insulation.J. Membrane Biol. 19:339–355
Johnson, R., Hammer, M., Sheridan, J., Revel, J.-P. 1974. Gap junction formation between reaggregated Novikoff hepatoma cells.Proc. Natl. Acad. sci. USA 71:4536–4543
Krebs, E. G. 1972. Protein kinases.Curr. Topics Cell. Reg. 5:99–133
Kuo, J. F., Greengard, P. 1969. Cyclic nucleotide-dependent protein kinases. IV. Widespread occurrence of adenosine 3′,5′-monophosphate-dependent protein kinase in various tissues and phyla of the animal kingdom.Proc. Natl. Acad. Sci. USA 64:1349–1355
Lawrence, T. S., Beers, W. H., Gilula, N. B. 1978. Transmission of hormonal stimulation by cell-to-cell communication.Nature (London) 272:501–506
Loewenstein, W. R. 1966. Permeability of membrane junctions.Ann. N. Y. Acad. Sci. 137:441–472
Loewenstein, W. R. 1979. Junctional intercellular communication and the control of growth.Biochim. Biophys. Acta Cancer Rev. 560:1–65
Loewenstein, W. R. 1981. Junctional intercellular communication. The cell-to-cell membrane channel.Physiol. Rev 61(4)
Loewenstein, W. R., Kanno, Y., Socolar, S. J. 1978. The cell-to-cell channel.Fed. Proc (Symp.) 37:2545–2650
Nimmo, H. G., Cohen, P. 1977. Hormonal control of protein phosphorylation.In: Advances in Cyclic Nucleotide Research. P. Greengard and G. A. Robison, editors. pp. 145–266. Raven Press, New York
Pauli, B. U., Weinstein, R., Soble, L. W., Alroy, J. 1977. Freezefracture of monolayer cultures.J. Cell Biol. 72:763–769
Pitts, J. D. 1977. Direct communication between animal cells.In: International Cell Biology 1976–1977. B. R. Brinkley and K. R. Porter, editors. pp. 43–49. Rockefeller University Press, New York
Revel, J. P., Yancey, S. B., Meyer, D. J., Nicholson, B. 1980. Cell junctions and intercellular communication.In Vitro 16(12): 1010–1017
Revel, J. P., Yee, A. G., Hudspeth, A. J. 1971. Gap junctions between electrotonically coupled cells.Proc. Natl. Acad. Sci. USA 68:2924–2938
Robison, G. A., Butcher, R. W., Sutherland, E. W. 1971. Cyclic AMP. Academic Press, New York
Schwarzmann, G. O. H., Wiegandt, H., Rose, B., Zimmerman, A., Ben-Haim, D., Loewenstein, W. R. 1981. Diameter of the cell-to-cell junctional membrane channels, as probed with neutral molecules.Science 213:551–553
Simpson, I., Rose, B., Loewenstein, W. R. 1979. Size limit of molecules permeating the junctional membrane channels.Science 195:294–296
Socolar, S. J., Loewenstein, W. R. 1979. Methods for studying transmission through permeable cell-to-cell junctions.In: Methods in Membrane Biology. E. Korn, editor. pp. 121–177. Plenum, New York
Tsien, R. W., Weingart, R. 1976. Inotropic effect of cyclic AMP in calf ventricular muscle studied by a cut end method.J. Physiol (London) 260:117–141
Unwin, P. M. T., Zampighi, G. 1980. Structure of the junction between communicating cells.Nature (London) 283:545–549
Vogt, M., Dulbecco, R. 1960. Virus-cell interaction with a tumor-producing virus.Proc. Natl. Acad. Sci. USA 46:365
Williams, E. H., DeHaan, R. L. 1981. Electrical coupling among heart cells in absence of ultrastructurally defined gap junctions.J. Membrane Biol. 60:237–248
Yancey, S. B., Easter, D., Revel, J.-P. 1979. Changes in gap junctions during liver regeneration.J. Ultrastruct. Res. 67:229–242
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Flagg-Newton, J.L., Dahl, G. & Loewenstein, W.R. Cell junction and cyclic AMP: I. Upregulation of junctional membrane permeability and junctional membrane particles by administration of cyclic nucleotide or phosphodiesterase inhibitor. J. Membrain Biol. 63, 105–121 (1981). https://doi.org/10.1007/BF01969452
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DOI: https://doi.org/10.1007/BF01969452