Advertisement

Pflügers Archiv

, Volume 429, Issue 6, pp 825–831 | Cite as

Phospholipase C activates protein kinase C during induction of slow Na current in Xenopus oocytes

  • Gilles Charpentier
  • Nathalie Béhue
  • Franck Fournier
Original Article Neurophysiology, Muscle and Sensory Organs

Abstract

Protein phosphorylation by protein kinase C (PKC) has recently been shown to be a key event in the induction of the slow inward Na current observed during sustained depolarization of the Xenopus oocyte membrane. The present work investigates the possible pathways leading to PKC activation. PKC is activated by a series of phospholipid metabolites, such as diacylglycerol (DAG) and arachidonic acid produced by phospholipases C (PLC) and A2(PLA2) respectively. To test whether PKC activation was dependent upon the phospholipid metabolites produced either by PLC or by PLA2, enzyme activity was reduced using selective inhibitors. Results indicated that inhibition of PLA2 activity and inhibition of the enzymes involved in the arachidonic acid cascade failed to affect Na current amplitude. On the other hand, PLC inhibition caused a marked decrease of Na current amplitude. In another series of experiments, Na current was fully restored, in spite of PLC inhibition, by directly enhancing PKC activity with a powerful activator phorbol 12-myristate 13-acetate. These data strongly suggest that PLC is involved in PKC activation during Na channel induction.

Keywords

Xenopus oocytes Sodium channels Phospholipase C Phospholipase A2 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Artalejo CR, Rossie S, Perlman RL, Fox AP (1992) Voltage-dependent phosphorylation may recruit Ca2+ current facilitation in chromaffin cells. Nature 358:63–66Google Scholar
  2. 2.
    Banga HS, Simons ER, Brass LF, Rittenhouse SE, (1986) Activation of phospholipases A and C in human platelets exposed to epinephrine: role of glycoproteins IIb/IIIa and dual role of epinephrine. Proc Natl Acad Sci USA 83:9197–9201Google Scholar
  3. 3.
    Baud C (1983) Developmental change of a depolarization-induced sodium permeability in the oocyte of Xenopus laevis. Dev Biol 99:524–528Google Scholar
  4. 4.
    Baud C, Kado RT (1984) Induction and disappearance of excitability in the oocyte of Xenopus laevis: a voltage-clamp study. J Physiol (Lond) 356:275–289Google Scholar
  5. 5.
    Baud C, Kado RT, Marcher K (1982) Sodium channels induced by depolarization of the Xenopus laevis oocyte. Proc Natl Acad Sci USA 79:3188–3192Google Scholar
  6. 6.
    Berridge MJ (1993) Inositol trisphosphate and calcium signalling. Nature 361:315–325Google Scholar
  7. 7.
    Bleasdale JE, Thakur NR, Gremban RS, Bundy GL, Fitzpatrick FA, Smith RJ, Bunting S (1990) Selective inhibition of receptor-coupled phospholipase C-dependent processes in human platelets and polymorphonuclear neutrophils. J Pharmacol Exp Ther 255:756–768Google Scholar
  8. 8.
    Castagna M, Takai Y, Kaibuchi K, Sano K, Kikkawa U, Nishizuka Y (1982) Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem 257:7847–7851Google Scholar
  9. 9.
    Charpentier G, Fournier F, Béhue N, Marlot D, Brûlé G (1993) Positive regulation by protein kinase C of slow Na current in Xenopus oocytes. Proc R Soc Lond [Biol] 254:15–20Google Scholar
  10. 10.
    Dumont JN (1972) Oogenesis in Xenopus laevis (Daudin). I. Stages of oocytes development in laboratory maintained animals. J Morphol 136:153–180Google Scholar
  11. 11.
    Gabev E, Kasianowicz J, Abbott T, McLaughlin S (1989) Binding of neomycin to phosphatidylinositol 4,5-bisphosphate (PIP2). Biochim Biophys Acta 979:105–112Google Scholar
  12. 12.
    Hawkins DJ, Brash AR (1989) Lipoxygenase metabolism of polyunsaturated fatty acids in oocytes of the frog Xenopus laevis. Arch Biochem Biophys 268:447–455Google Scholar
  13. 13.
    Huang KP (1989) The mechanism of protein kinase C activation. Trends Neurosci 12:425–432Google Scholar
  14. 14.
    Kado RT, Baud C (1981) The rise and fall of electrical excitability in the oocyte of Xenopus laevis. J Physiol (Paris) 77:1113–1117Google Scholar
  15. 15.
    Kado RT, Marcher K, Ozon R (1979) Mise en évidence d'une dépolarisation de longue durée dans l'ovocyte de Xenopus laevis. CR Acad Sci III 288:1187–1189Google Scholar
  16. 16.
    Leonard JP, Nargeot J, Snutch TP, Davidson N, Lester HA (1987) Ca channels induced in Xenopus oocytes by rat brain mRNA. J Neurosci 7:875–881Google Scholar
  17. 17.
    Lodhi S, Weiner ND, Schacht J (1979) Interactions of neomycin and calcium in synaptosomal membranes and polyphosphoinositide monolayers. Biochim Biophs Acta 557:1–8Google Scholar
  18. 18.
    Madi S, Giessler J, Hirschelmann R, Friedrich G, Braquet P (1991) Allergen-induced bronchospasm in passively sensitized guinea pigs: influence of new substances in comparison to reference compounds. Agents Actions 32:144–145Google Scholar
  19. 19.
    Nishizuka Y (1986) Studies and perspectives of protein kinase C. Science 233:305–312Google Scholar
  20. 20.
    Nishizuka Y (1988) The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature 334:661–665Google Scholar
  21. 21.
    Parekh AB, Terlau H, Stühmer W (1993) Depletion of InsP3 stores activates a Ca2+ and K+ current by means of a phosphatase and a diffusible messenger. Nature 364:814–818Google Scholar
  22. 22.
    Piomelli D, Greengard P (1990) Lipoxygenase metabolites of arachidonic acid in neuronal transmembrane signalling. Trends Pharmacol Sci 11:367–373Google Scholar
  23. 23.
    Rhee SG, Choi KD (1992) Multiple forms of phospholipase C isozymes and their activation mechanisms. Adv Second Messenger Phosphoprotein Res 26:35–61Google Scholar
  24. 24.
    Rock CO, Jackowski S (1987) Thrombin- and nucleotide-activated phosphatidylinositol 4,5-bisphosphate phospholipase C in human platelet membranes. J Biol Chem 262:5492–5498Google Scholar
  25. 25.
    Sculptoreanu A, Scheuer T, Catterall WA (1993) Voltage-dependent potentiation of L-type Ca2+ channels due to phosphorylation by cAMP-dependent protein kinase. Nature 364:240–243Google Scholar
  26. 26.
    Smith RJ, Sam LM, Justen JM, Bundy GL, Bala GA, Bleasdale JE (1990) Receptor-coupled signal transduction in human polymorphonuclear neutrophils: effects of a novel inhibitor of phospholipase C-dependent processes on cell responsiveness. J Pharmacol Exp Ther 253:688–697Google Scholar
  27. 27.
    Umbach JA Gundersen CB (1987) Expression of an ω-conotoxin-sensitive calcium channel in Xenopus oocytes injected with mRNA from Torpedo electric lobe. Proc Natl Acad Sci USA 84:5464–5468Google Scholar
  28. 28.
    Vasilets LA, Schmalzing G, Madefessel K, Haase W, Schwarz W, (1987) Activation of protein kinase C by phorbol ester induces down regulation of the Na+/K+-ATPase in the oocyte of Xenopus laevis. J Membr Biol 188:131–142Google Scholar
  29. 29.
    Vergara J, Tsien RY, Delay M (1985) Inositol 1,4,5-trisphosphate: a possible chemical link in excitation-contraction coupling in muscle. Proc Natl Acad Sci USA 82:6352–6356Google Scholar
  30. 30.
    Whitaker MJ, Aitchison MJ (1985) Calcium-dependent polyphosphoinositide hydrolysis is associated with exocytosis in vitro. FEBS Lett 182:119–124Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Gilles Charpentier
    • 1
  • Nathalie Béhue
    • 1
  • Franck Fournier
    • 1
  1. 1.Laboratoire de Neurobiologie CellulaireUniversité de Picardie Jules Verne, Faculté des SciencesAmiens cedex 1France

Personalised recommendations