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. 1991 Aug 1;114(3):455–464. doi: 10.1083/jcb.114.3.455

The polarized distribution of poly(A+)-mRNA-induced functional ion channels in the Xenopus oocyte plasma membrane is prevented by anticytoskeletal drugs

PMCID: PMC2289088  PMID: 1713591

Abstract

Foreign mRNA was expressed in Xenopus laevis oocytes. Newly expressed ion currents localized in defined plasma membrane areas were measured using the two-electrode voltage clamp technique in combination with a specially designed chamber, that exposed only part of the surface on the oocytes to channel agonists or inhibitors. Newly expressed currents were found to be unequally distributed in the surface membrane of the oocyte. This asymmetry was most pronounced during the early phase of expression, when channels could almost exclusively be detected in the animal hemisphere of the oocyte. 4 d after injection of the mRNA, or later, channels could be found at a threefold higher density at the animal than at the vegetal pole area. The pattern of distribution was observed to be similar with various ion channels expressed from crude tissue mRNA and from cRNAs coding for rat GABAA receptor channel subunits. Electron microscopical analysis revealed very similar microvilli patterns at both oocyte pole areas. Thus, the asymmetric current distribution is not due to asymmetric surface structure. Upon incubation during the expression period in either colchicine or cytochalasin D, the current density was found to be equal in both pole areas. The inactive control substance beta-lumicolchicine had no effect on the asymmetry of distribution. Colchicine was without effect on the amplitude of the expressed whole cell current. Our measurements reveal a pathway for plasma membrane protein expression endogenous to the Xenopus oocyte, that may contribute to the formation and maintenance of polarity of this highly organized cell.

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Selected References

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  1. Achler C., Filmer D., Merte C., Drenckhahn D. Role of microtubules in polarized delivery of apical membrane proteins to the brush border of the intestinal epithelium. J Cell Biol. 1989 Jul;109(1):179–189. doi: 10.1083/jcb.109.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barnard E. A., Miledi R., Sumikawa K. Translation of exogenous messenger RNA coding for nicotinic acetylcholine receptors produces functional receptors in Xenopus oocytes. Proc R Soc Lond B Biol Sci. 1982 May 22;215(1199):241–246. doi: 10.1098/rspb.1982.0040. [DOI] [PubMed] [Google Scholar]
  3. Bartles J. R., Hubbard A. L. Plasma membrane protein sorting in epithelial cells: do secretory pathways hold the key? Trends Biochem Sci. 1988 May;13(5):181–184. doi: 10.1016/0968-0004(88)90147-8. [DOI] [PubMed] [Google Scholar]
  4. Cathala G., Savouret J. F., Mendez B., West B. L., Karin M., Martial J. A., Baxter J. D. A method for isolation of intact, translationally active ribonucleic acid. DNA. 1983;2(4):329–335. doi: 10.1089/dna.1983.2.329. [DOI] [PubMed] [Google Scholar]
  5. Ceriotti A., Colman A. Binding to membrane proteins within the endoplasmic reticulum cannot explain the retention of the glucose-regulated protein GRP78 in Xenopus oocytes. EMBO J. 1988 Mar;7(3):633–638. doi: 10.1002/j.1460-2075.1988.tb02857.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Colman A., Jones E. A., Heasman J. Meiotic maturation in Xenopus oocytes: a link between the cessation of protein secretion and the polarized disappearance of Golgi apparati. J Cell Biol. 1985 Jul;101(1):313–318. doi: 10.1083/jcb.101.1.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Colman A., Morser J., Lane C., Besley J., Wylie C., Valle G. Fate of secretory proteins trapped in oocytes of Xenopus laevis by disruption of the cytoskeleton or by imbalanced subunit synthesis. J Cell Biol. 1981 Dec;91(3 Pt 1):770–780. doi: 10.1083/jcb.91.3.770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cooper J. A. Effects of cytochalasin and phalloidin on actin. J Cell Biol. 1987 Oct;105(4):1473–1478. doi: 10.1083/jcb.105.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dawid I. B., Sargent T. D. Xenopus laevis in developmental and molecular biology. Science. 1988 Jun 10;240(4858):1443–1448. doi: 10.1126/science.3287620. [DOI] [PubMed] [Google Scholar]
  10. Dotti C. G., Simons K. Polarized sorting of viral glycoproteins to the axon and dendrites of hippocampal neurons in culture. Cell. 1990 Jul 13;62(1):63–72. doi: 10.1016/0092-8674(90)90240-f. [DOI] [PubMed] [Google Scholar]
  11. Dreyfus P. A., Seidman S., Pincon-Raymond M., Murawsky M., Rieger F., Schejter E., Zakut H., Soreq H. Tissue-specific processing and polarized compartmentalization of clone-produced cholinesterase in microinjected Xenopus oocytes. Cell Mol Neurobiol. 1989 Sep;9(3):323–341. doi: 10.1007/BF00711413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Drummond D. R., McCrae M. A., Colman A. Stability and movement of mRNAs and their encoded proteins in Xenopus oocytes. J Cell Biol. 1985 Apr;100(4):1148–1156. doi: 10.1083/jcb.100.4.1148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Durand-Schneider A. M., Maurice M., Dumont M., Feldmann G. Effect of colchicine and phalloidin on the distribution of three plasma membrane antigens in rat hepatocytes: comparison with bile duct ligation. Hepatology. 1987 Nov-Dec;7(6):1239–1248. doi: 10.1002/hep.1840070611. [DOI] [PubMed] [Google Scholar]
  14. Lane C. D. The fate of genes, messengers, and proteins introduced into Xenopus oocytes. Curr Top Dev Biol. 1983;18:89–116. doi: 10.1016/s0070-2153(08)60580-3. [DOI] [PubMed] [Google Scholar]
  15. Malherbe P., Draguhn A., Multhaup G., Beyreuther K., Möhler H. GABAA-receptor expressed from rat brain alpha- and beta-subunit cDNAs displays potentiation by benzodiazepine receptor ligands. Brain Res Mol Brain Res. 1990 Aug;8(3):199–208. doi: 10.1016/0169-328x(90)90017-8. [DOI] [PubMed] [Google Scholar]
  16. Matter K., Bucher K., Hauri H. P. Microtubule perturbation retards both the direct and the indirect apical pathway but does not affect sorting of plasma membrane proteins in intestinal epithelial cells (Caco-2). EMBO J. 1990 Oct;9(10):3163–3170. doi: 10.1002/j.1460-2075.1990.tb07514.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Matus-Leibovitch N., Lupu-Meiri M., Oron Y. Two types of intrinsic muscarinic responses in Xenopus oocytes. II. Hemispheric asymmetry of responses and receptor distribution. Pflugers Arch. 1990 Oct;417(2):194–199. doi: 10.1007/BF00370699. [DOI] [PubMed] [Google Scholar]
  18. Methfessel C., Witzemann V., Takahashi T., Mishina M., Numa S., Sakmann B. Patch clamp measurements on Xenopus laevis oocytes: currents through endogenous channels and implanted acetylcholine receptor and sodium channels. Pflugers Arch. 1986 Dec;407(6):577–588. doi: 10.1007/BF00582635. [DOI] [PubMed] [Google Scholar]
  19. Oron Y., Gillo B., Gershengorn M. C. Differences in receptor-evoked membrane electrical responses in native and mRNA-injected Xenopus oocytes. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3820–3824. doi: 10.1073/pnas.85.11.3820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Poo M. Rapid lateral diffusion of functional A Ch receptors in embryonic muscle cell membrane. Nature. 1982 Jan 28;295(5847):332–334. doi: 10.1038/295332a0. [DOI] [PubMed] [Google Scholar]
  21. Rindler M. J., Ivanov I. E., Sabatini D. D. Microtubule-acting drugs lead to the nonpolarized delivery of the influenza hemagglutinin to the cell surface of polarized Madin-Darby canine kidney cells. J Cell Biol. 1987 Feb;104(2):231–241. doi: 10.1083/jcb.104.2.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Robinson K. R. Electrical currents through full-grown and maturing Xenopus oocytes. Proc Natl Acad Sci U S A. 1979 Feb;76(2):837–841. doi: 10.1073/pnas.76.2.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Salas P. J., Misek D. E., Vega-Salas D. E., Gundersen D., Cereijido M., Rodriguez-Boulan E. Microtubules and actin filaments are not critically involved in the biogenesis of epithelial cell surface polarity. J Cell Biol. 1986 May;102(5):1853–1867. doi: 10.1083/jcb.102.5.1853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sigel E., Baur R. Activation of protein kinase C differentially modulates neuronal Na+, Ca2+, and gamma-aminobutyrate type A channels. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6192–6196. doi: 10.1073/pnas.85.16.6192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sigel E. Effects of veratridine on single neuronal sodium channels expressed in Xenopus oocytes. Pflugers Arch. 1987 Sep;410(1-2):112–120. doi: 10.1007/BF00581903. [DOI] [PubMed] [Google Scholar]
  26. Sigel E. Use of Xenopus oocytes for the functional expression of plasma membrane proteins. J Membr Biol. 1990 Sep;117(3):201–221. doi: 10.1007/BF01868451. [DOI] [PubMed] [Google Scholar]
  27. Simons K., Wandinger-Ness A. Polarized sorting in epithelia. Cell. 1990 Jul 27;62(2):207–210. doi: 10.1016/0092-8674(90)90357-k. [DOI] [PubMed] [Google Scholar]
  28. Soreq H. The biosynthesis of biologically active proteins in mRNA-microinjected Xenopus oocytes. CRC Crit Rev Biochem. 1985;18(3):199–238. doi: 10.3109/10409238509085134. [DOI] [PubMed] [Google Scholar]
  29. Strous G. J., Willemsen R., van Kerkhof P., Slot J. W., Geuze H. J., Lodish H. F. Vesicular stomatitis virus glycoprotein, albumin, and transferrin are transported to the cell surface via the same Golgi vesicles. J Cell Biol. 1983 Dec;97(6):1815–1822. doi: 10.1083/jcb.97.6.1815. [DOI] [PMC free article] [PubMed] [Google Scholar]

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