Skip to main content
SpringerLink
Log in

Diacylglycerol-induced activation of protein kinase C attenuates Na+ currents by enhancing inactivation from the closed state

  • Original Article
  • Molecular and Cellular Physiology
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

The causes of attenuation of Na+ currents by diacylglycerol (DAG)-induced protein kinase C (PKC) activation in mouse neuroblastoma N1E-115 cells were investigated using the cell-attached patch, and the perforated-patch (nystatin based) whole-cell voltage-clamp techniques. Activation of PKC by DAG attenuated Na+ currents. Attenuation occurred in the absence of significant changes in the time-course of Na+ currents. However, the steady-state inactivation curve of these currents shifted to more negative voltages by approximately 20 mV. Here we demonstrate that the time-course of inactivation is accelerated by treatment with DAG-like substances in a voltage-dependent manner (time constant of inactivation decreased by 2- and 3.6-fold at −60, and −30 mV, respectively). In cell-attached patches, treatment with DAG compounds increased the percentage of current traces showing no single Na+ channel openings in response to depolarizing voltage-clamp pulses. Moreover, the average of current traces containing single Na+ channel openings was essentially the same in control conditions and after treatment with DAG compounds. Removal of Na+ channel inactivation by the alkaloid batrachotoxin prevented the attenuation of Na+ currents by PKC activation via DAGs. Taken together, these data strongly suggest that PKC-induced attenuation of Na+ currents is linked to an enhancement of Na+ channel inactivation. This attenuation is caused by an increase in the number of Na+ channels inactivating directly from the closed state(s). This inactivation pathway represents a simple and efficient physiological mechanism by which PKC activation might modulate the electrical activity of excitable cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Aldrich RW, Stevens CF (1983) Inactivation of open and closed sodium channels determined separately. Cold Spring Harb Symp Quant Biol 48:147–153

    Google Scholar 

  2. Aldrich RW, Corey DP, Stevens CF (1983) A reinterpretation of mammalian sodium channel gating based on single-channel recording. Nature 306:436–441

    Google Scholar 

  3. Armstrong CM, Bezanilla F (1977) Inactivation of the sodium channel. II. Gating current experiments. J Gen Physiol 70:567–590

    Google Scholar 

  4. Bean BP (1981) Sodium channel inactivation in crayfish giant axon. Must channels open before inactivating? Biophys J 35:595–614

    Google Scholar 

  5. Benz I, Herzig JW, Kohlhardt M (1992) Opposite effects of angiotensin II and the protein kinase C activator OAG on cardiac Na+ channels. J Membr Biol 130:183–190

    Google Scholar 

  6. Bezanilla F, Armstrong CM (1977) Inactivation of the sodium channel. I. Sodium current experiments. J Gen Physiol 70:549–566

    Google Scholar 

  7. Catterall WA (1992) Cellular, molecular biology of voltagegated sodium channels. Physiol Rev 72:S15-S48

    Google Scholar 

  8. Costa MR, Catterall WA (1984) Phosphorylation of the alpha subunit of the sodium channel by protein kinase C. Cell Mol Neurobiol 4:291–297

    Google Scholar 

  9. Cukierman S (1991) Inactivation modifiers of Na currents and the gating of rat brain sodium channels in planar lipid membranes. Pflügers Arch 419:514–521

    Google Scholar 

  10. Cukierman S (1992) Characterization of K+ currents in rat malignant lymphocytes (Nb2 cells). J Membr Biol 126:147–157

    Google Scholar 

  11. Dascal N, Lotan I (1991) Activation of protein kinase C alters voltage dependence of a Na+ channel. Neuron 6:165–175

    Google Scholar 

  12. Emerick MC, Agnew WS (1989) Identification of phosphorylation sites for adenosine 3′, 5′-cyclic phosphate dependent protein kinase on the voltage-sensitive sodium channel from Electrophorus electricus. Biochemistry 28:8367–8380

    Google Scholar 

  13. Godoy CM, Cukierman S (1994) Multiple effects of protein kinase C activators on Na+ channels in mouse neuroblastoma cells. J Membr Biol 140:101–110

    Google Scholar 

  14. Hille B (1992) Ionic channels of excitable membranes. Sinauer, Sunderland Mass.

    Google Scholar 

  15. Hodgkin AL, Huxley AF (1952) A quantitative decription of membrane current and its application to conduction and excitation in nerve. J Physiol (Lond) 117:500–544

    Google Scholar 

  16. Horn R, Marty A (1988) Muscarinic activation of ionic currents measured by a new whole-cell recording method. J Gen Physiol 92:145–159

    Google Scholar 

  17. Huang LYM, Moran N, Ehrenstein G (1984) Gating kinetics of batrachotoxin-modified sodium channels in neuroblastoma cells determined from single-channel measurements. Biophys 145:313–321

    Google Scholar 

  18. Kaczmarek LK, Levitan IB (1987) Neuromodulation. The biochemical control of neuronal excitability. Oxford University Press, New York

    Google Scholar 

  19. Khodorov BI, Revenko SV (1979) Further analysis of the mechanism of action of batrachotoxin on the membrane of the myelinated nerve. Neuroscience 4:1315–1330

    Google Scholar 

  20. Korn SJ, Marty A, Connor JA, Horn R (1991) Perforated patch recording. Methods Neurosci 4:264–373

    Google Scholar 

  21. Li M, West JW, Numann R, Murphy BJ, Scheuer TL, Catterall WA (1993) Convergent regulation of sodium channels by protein kinase C and cAMP-dependent protein kinase. Science 261:1439–1442

    Google Scholar 

  22. Linden DJ, Routtenberg A (1989) Cis-fatty acids, which activate protein kinase C, attenuate Na+ and Ca2+ currents in mouse neuroblastoma cells. J Physiol (Lond) 419:95–119

    Google Scholar 

  23. Lotan I, Dascal N, Naor Z, Boton R (1990) Modulation of vertebrate brain Na+ and K+ channels by subtypes of protein kinase C. FEBS Lett 267:25–28

    Google Scholar 

  24. Moorman JR, Kirsch GE, Brown AM (1989) Angiotensin II and phorbol ester modulate cardiac sodium channels in neonatal rat (abstract). Biophys J 55:229a

    Google Scholar 

  25. Murphy BJ, Catterall WA (1992) Phosphorylation of purified rat brain Na+ channel reconstituted into phospholipid vesicles by protein kinase C. J Biol Chem 267:16129–16134

    Google Scholar 

  26. Numann R, Catteral WA, Scheuer T (1991) Functional modulation of brain sodium channels by protein kinase C phosphorylation. Science 254:115–118

    Google Scholar 

  27. Qu Y, Rogers J, Tanada T, Scheuer T, Catterall WA (1994) Modulation of cardiac Na+ channels expressed in a mammalian cell line and in ventricular myocytes by protein kinase C. Proc Natl Acad Sci USA 91:3289–3293

    Google Scholar 

  28. Rack M, Rubly N, Waschow C (1986) Effects of some chemical reagents on sodium current inactivation in myelinated nerve fibers of the frog. Biophys J 50:557–564

    Google Scholar 

  29. Rossie S, Catterall WA (1987) Cyclic AMP-dependent phosphorylation of voltage-sensitive sodium channels in primary cultures of rat brain neurons. J Biol Chem 262:12735–12744

    Google Scholar 

  30. Sigel E, Baur R (1988) Activation of protein kinase C differentially modulated neuronal Na+, Ca2+, and gamma-aminobutyrate type A channels. Proc Natl Acad Sci USA 85:6192–6196

    Google Scholar 

  31. Tanguy J, Yeh JZ (1991) BTX modification of Na channels in squid axons. I. State dependence of BTX action. J Gen Physiol 97:499–520

    Google Scholar 

  32. Vandenberg C, Horn R (1984) Inactivation viewed through single sodium channels. J Gen Physiol 84:535–564

    Google Scholar 

  33. West JW, Numann R, Murphy BJ, Scheuer T, Catterall WA (1991) A phosphorylation site in the Na+ channel required for modulation by protein kinase C. Science 254:866–868

    Google Scholar 

  34. Yang J, Barchi R (1990) Phosphorylation of the rat skeletal muscle sodium channel by cyclic AMP-dependent protein kinase. J Neurochem 54:954–962

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Physiology, Loyola University Medical Center, 2160 South First Avenue, 60153, Maywood, Ill, USA

    Carlos Marcelo G. Godoy & Samuel Cukierman

Authors
  1. Carlos Marcelo G. Godoy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Samuel Cukierman
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Godoy, C.M.G., Cukierman, S. Diacylglycerol-induced activation of protein kinase C attenuates Na+ currents by enhancing inactivation from the closed state. Pflugers Arch. 429, 245–252 (1994). https://doi.org/10.1007/BF00374319

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00374319

Key words

Search

Navigation