Summary
mRNA from estrogen-stimulated rat myometrium, a tissue known to upregulate cell-cell channels in response to this hormone, was microinjected intoXenopus laevis oocytes. The oocytes had been freed from covering layers of follicle cells and vitelline to allow direct cell membrane interactions when paired. About 4 hours after the mRNA injection, paired oocytes become electrically coupled. This coupling was due to the presence of typical cell-cell channels characterized by size-limited intercellular tracer flux, the presence of gap junctions at the oocyte-oocyte interface, and the reversible uncoupling that occurred in the presence of carbon dioxide. The induction of new cell-cell channels in the oocyte membrane was observed against a zero background or a low level of endogenous coupling, depending on the maturation stage of the oocytes. The time course of development of cell-cell coupling after the microinjection of mRNA was determined. The mRNA capable of inducing cell-cell coupling was confined to an intermediate size class when fractionated on a sucrose gradient.
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Azarnia, R., Dahl, G., Loewenstein, W.R. 1981. Cell junction and cyclic AMP: III. Promotion of junctional membrane permeability and junctional membrane particles in a junction-deficient cell-type.J. Membrane Biol. 63:133–146
Browne, C.L., Wiley, H.S., Dumont, J.N. 1979. Oocyte-follicle cell gap junctions inXenopus laevis and the effects of gonadotropin on their permeability.Science 203:182–183
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
Chao, N., Young, S.H., Poo, M. 1981. Localization of cell membrane components by surface diffusion into a “trap”.Biophys. J. 36:139–153
Chirgwin, J.M., Przybyla, A.E., MacDonald, R.J., Rutter, W.J. 1979. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.Biochemistry 18:5294–5299
Dahl, G., Azarnia, R., Werner, R. 1980. De novo construction of cell-to-cell channels.In Vitro 16:1068–1075
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
Dumont, J.N. 1972. Oogenesis inXenopus laevis (Daudin).J. Morphol. 136:153–180
Fallon, R.F., Goodenough, D.A. 1981. Five-hour half-life of mouse liver gap junction protein.J. Cell Biol. 90:521–526
Finbow, M., Yancey, S.B., Johnson, R., Revel, J.P. 1980. Independent lines of evidence suggesting a major gap junctional protein with a molecular weight of 26,000.Proc. Natl. Acad. Sci. USA 77:970–974
Garfield, R.E., Kanaan, M.S., Daniel, E.E. 1980. Gap junction formation in myometrium: Control by estrogens, progesterone, and prostaglandins.Am. J. Physiol. 238:C81-C89
Gilula, N.B., Epstein, M.L., Beers, W.H. 1978. Cell-to-cell communication and ovulation: A study of the cumulus-oocyte complex.J. Cell Biol. 78:58–75
Goodenough, D.A. 1976. In vitro formation of gap junctional vesicles.J. Cell Biol. 68:220–232
Gundersen, C.B., Miledi, R., Parker, I. 1984. Messenger RNA from human brain induces drug- and voltage-operated channels inXenopus oocytes.Nature (London) 308:421–424
Gurdon, J.B., Lingrel, J.B., Marbaiz, G. 1973. Message stability in infected frog oocytes: Long life of mammalian alpha and beta globulin messages.J. Mol. Biol. 80:539–551
Henderson, D., Eibl, H., Weber, K. 1979. Structure and biochemistry of mouse hepatic gap junctions.J. Mol. Biol. 132:193–218
Henderson, D., Weber, K. 1982. Immunological analysis of gap junction proteins from liver, lens and heart muscle.Biol. Cell 45:229a
Hertzberg, E.C., Gilula, N.B. 1978. Isolation and characterization of gap junctions from rat liver.J. Biol. Chem. 254:2138–2147
Kusano, K., Miledi, R., Stinnakre, J. 1982. Cholinergic and cate-cholaminergic receptors in theXenopus oocyte membrane.J. Physiol. (London) 328:143–170
Lane, C., Knowland, J. 1975. The injection of mRNA into living cells: The use of frog oocytes for the assay of mRNA in the study of the control of gene expression.In: The Biochemistry of Animal Development. R. Weber, editor. Vol. 3, pp. 145–181. Academic, New York
Loewenstein, W.R. 1981. Junctional intercellular communication: The cell-to-cell membrane channel.Physiol. Rev. 61:829–912
Makowski, L., Caspar, D.L.C., Phillips, W.C., Goodenough, D.A. 1977. Gap junction structures: II. Analysis of the X-ray diffraction data.J. Cell Biol. 74:629–645
Manjunath, C.K., Goings, G.E., Page, E. 1984. Cytoplasmic surface and intramembrane components of rat heart gap junctional proteins.Am. J. Physiol. 246:H865-H875
Miledi, R., Parker, I., Sumikawa, K. 1982. Properties of acetylcholine receptors translated by cat muscle mRNA inXenopus oocytes.EMBO J 1:1307–1312
Mishina, M., Kurosaki, T., Tobimatsu, T., Morimoto, Y., Noda, M., Yamamoto, T., Terao, M., Lindstrom, J., Takahashi, T., Kuno, M., Numa, S. 1984. Expression of functional acetylcholine receptor from cloned cDNAs.Nature (London) 307:604–608
Noda, M., Shimizu, S., Tanabe, T., Takai, T., Kayano, T., Ikeda, T., Takahashi, H., Nakayama, H., Kanaoka, Y., Minamino, N., Kangawa, K., Matsuo, H., Raftery, M.A., Hirose, T., Inayama, S., Hayashida, H., Miyata, T., Numa, S. 1984. Primary structure ofElectrophorus electricus sodium channel deduced from cDNA sequence.Nature (London) 312:121–127
Preus, D., Johnson, A., Sheridan, J. 1981. Gap junctions between Novikoff hepatoma cells following dissociation and recovery in the absence of cell contact.J. Ultrastruct. Res. 77:248–262
Rose, B., Rick, R. 1978. Intracellular pH, intracellular free Ca, and junctional cell-cell coupling.J. Membrane Biol. 44:377–415
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. Vol. 10, pp. 121–177. Plenum, New York
Spray, D.C., Harris, A.L., Bennett, M.V.L. 1981. Gap junctional conductance is a simple and sensitive function of intracellular pH.Science 211:712–714
Suzuki, Y., Gage, L.P., Brown, D.D. 1972. The genes for silk fibroin inBombyx mori.J. Mol. Biol. 70:637–649
Turin, L., Warner, A. 1977. Carbon dioxide reversibly abolishes ionic communication between cells of early amphibian embryo.Nature (London) 270:56–57
Unwin, P.N.T., Zampighi, G. 1980. Structure of the junction between communicating cells.Nature (London) 283: 545–549
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Werner, R., Miller, T., Azarnia, R. et al. Translation and functional expression of cell-cell channel mRNA inXenopus oocytes. J. Membrain Biol. 87, 253–268 (1985). https://doi.org/10.1007/BF01871226
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DOI: https://doi.org/10.1007/BF01871226