Skip to main content
SpringerLink
Log in

Inactivation of the Ba2+ current in dissociated Helix neurons: voltage dependence and the role of phosphorylation

  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

The rate of inactivation of the voltage-dependent Ba2+ current in dissociated neurons from the snail Helix aspersa was found to be modulated by phosphorylation. Conditions were chosen such that the most likely mechanism of inactivation of the Ba2+ current was a voltage-dependent/calcium-independent inactivation process. If adenosine-triphosphate (ATP) was not included in the patch electrode filling solution, or if alkaline phosphatase was added, the Ba2+ current rapidly ran down and the rate of inactivation greatly increased with time. Dialysis with either ATPγS or the phosphatase inhibitor okadaic acid (OA) either enhanced the amplitude or greatly reduced the rate of run-down of the Ba2+ current (depending upon the presence of ATP), as well as reducing the rate of inactivation. However, dialysis with either the catalytic subunit of the cyclic-adenosine-monophosphate-dependent protein kinase (cAMP-PK), a synthetic peptide inhibitor of this enzyme, or staurosporine (a potent inhibitor of protein kinase C), did not have any significant effect on the amplitude or kinetics of the Ba2+ current. Surprisingly, dialysis with a peptide inhibitor (CKIP) of the Ca2+/calmodulin-dependent protein kinase II (Ca2+-CaM-PK) significantly reduced the rate of inactivation of this current. These results suggest that phosphorylation may exert its effect by modulating the gating properties of the Ca2+ channels.

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. Armstrong DL (1989) Calcium channel regulation by calcineurin, a Ca2+-activated phosphatase in mammalian brain. Trends Neurosci 12:117–122

    Google Scholar 

  2. Armstrong, D, Eckert R (1987) Voltage-activated calcium channels that must be phosphorylated to respond to membrane depolarization. Proc Natl Acad Sci USA 84:2518–2522

    Google Scholar 

  3. Armstrong D, Erxleben C, Kalman D, Lai Y, Nairn A, Greengard P (1988) Intracellular calcium controls the activity of dihydropyridine-sensitive calcium channels through protein phosphorylation and its removal. J Gen Physiol 92:10a

    Google Scholar 

  4. Augustine CK, Bezanilla F (1990) Phosphorylation modulates potassium conductance and gating current of perfused giant axons of squid. J Gen Physiol 95:245–271

    Google Scholar 

  5. Brown AM, Morimoto K, Tsuda Y, Wilson DL (1981) Calcium current-dependent and voltage-dependent inactivation of calcium channels in Helix aspersa. J Physiol (Lond) 320:193–218

    Google Scholar 

  6. Chad JE, Eckert R (1986) An enzymatic mechanism for calcium current inactivation in dialysed Helix neurones. J Physiol (Lond) 378:31–51

    Google Scholar 

  7. Cheng H-C, Kemp BE, Pearson RB, Smith AJ, Misconi L, Van Patten SM, Walsh DA (1986) A potent synthetic peptide inhibitor of the cAMP-dependent protein kinase. J Biol Chem 261:989–992

    Google Scholar 

  8. Cohen P (1989) The structure and regulation of protein phosphatases. Annu Rev Biochem 58:453–508

    Google Scholar 

  9. Cohen P, Holmes CFB, Tsukitani Y (1990) Okadaic acid: a new probe for the study of cellular regulation. Trends Biochem Sci 15:98–102

    Google Scholar 

  10. DeRiemer SA, Strong JA, Albert KA, Greengard P, Kaczmarek LK (1985) Enhancement of calcium current in Aplysia neurones by phorbol ester and protein kinase C. Nature 313:313–316

    Google Scholar 

  11. Doroshenko PA, Kostyuk PG, Martynyuk AE (1982) Intracellular metabolism of adenosine 3′,5′-cyclic monophosphate and calcium inward current in perfused neurones of Helix pomatia. Neuroscience 7:2125–2134

    Google Scholar 

  12. Doroshenko PA, Kostyuk PG, Martynyuk AE, Kursky MD, Vorobetz ZD (1984) Intracellular protein kinase and calcium inward currents in perfused neurones of the snail Helix pomatia. Neuroscience 11:263–267

    Google Scholar 

  13. Eckert R, Chad JE (1984) Inactivation of calcium channels. Prog Biophys Mol Biol 44:215–267

    Google Scholar 

  14. Flockerzi V, Oeken H-J, Hofmann F, Pelzer D, Cavalié A, Trautwein W (1986) Purified dihydropyridine-binding site from skeletal muscle t-tubules is a functional calcium channel. Nature 323:66–68

    Google Scholar 

  15. Gerschenfeld HM, Paupardin-Tritsch D, Yakel JL (1991) Muscarinic enhancement of the voltage-dependent calcium current in an identified snail neuron. J Physiol (Lond) 434:85–105

    Google Scholar 

  16. Gratecos D, Fischer EH (1974) Adenosine 5′-O(3-thiotriphosphate) in the control of phosphorylase activity. Biochem Biophys Res Commun 58:960–967

    Google Scholar 

  17. Gross RA, Uhler MD, Macdonald RL (1990) The reduction of neuronal calcium currents by ATP-γ-S is mediated by a G protein and occurs independently of cyclic AMP-dependent protein kinase. Brain Res 535:214–220

    Google Scholar 

  18. Gutnick MJ, Lux HD, Swandulla D, Zucker H (1989) Voltage-dependent and calcium-dependent inactivation of calcium channel current in identified snail neurones. J Physiol (Lond) 412:197–220

    Google Scholar 

  19. Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100

    Google Scholar 

  20. Hammond C, Paupardin-Tritsch D, Nairn AC, Greengard P, Gerschenfeld HM (1987) Cholecystokinin induces a decrease in Ca2+ current in snail neurons that appears to be mediated by protein kinase C. Nature 325:809–811

    Google Scholar 

  21. Hartzell HC, Méry P-F, Fischmeister R, Szabo G (1991) Sympathetic regulation of cardiac calcium current is due exclusively to cAMP-dependent phosphorylation. Nature 351:573–576

    Google Scholar 

  22. Kalman D, O'Lague PH, Erxleben C, Armstrong DL (1988) Calcium-dependent inactivation of the dihydropyridine-sensitive calcium channels in GH3 cells. J Gen Physiol 92:531–548

    Google Scholar 

  23. Kameyama M, Hescheler J, Hofmann F, Trautwein W (1986) Modulation of Ca current during the phosphorylation cycle in the guinea pig heart. Pflügers Arch 407:123–128

    Google Scholar 

  24. Klee CB, Crouch TH, Krinks MH (1979) Calcineurin: A calcium and calmodulin-binding protein of the nervous system. Proc Natl Acad Sci USA 76:6270–6273

    Google Scholar 

  25. Klumpp, S, Cohen P, Schultz JE (1990) Okadaic acid, an inhibitor of protein phosphatase 1 in Paramecium, causes sustained Ca2+-dependent backward swimming in response to depolarizing stimuli. EMBO J 9:685–689

    Google Scholar 

  26. Nichols RA, Sihra TS, Czernik AJ, Nairn AC, Greengard P (1990) Calcium/calmodulin-dependent protein kinase II increases glutamate and noradrenaline release from synaptosomes. Nature 343:647–651

    Google Scholar 

  27. Otero AS, Breitwieser GE, Szabo G (1988) Activation of muscarinic potassium currents by ATPγS in atrial cells. Science 242:443–445

    Google Scholar 

  28. Palvimo J, Linnala-Kankkunen A, Mäenpää PH (1985) Thiophosphorylation and phosphorylation of chromatin proteins from calf thymus in vitro. Biochem Biophys Res Commun 126:103–108

    Google Scholar 

  29. Paupardin-Tritsch D, Hammond C, Gerschenfeld HM (1986) Serotonin and cyclic GMP both induce an increase of the calcium current in the same identified molluscan neurons. J Neurosci 6:2715–2723

    Google Scholar 

  30. Paupardin-Tritsch D, Hammond G, Gerschenfeld HM, Nairn AC, Greengard P (1986) cGMP-dependent protein kinase enhances Ca2+ current and potentiates the serotonin-induced Ca2+ current increase in snail neurones. Nature 323:812–814

    Google Scholar 

  31. Perozo E, Bezanilla F (1990) Phosphorylation affects voltage gating of the delayed rectifier K+ channel by electrostatic interactions. Neuron 5:685–690

    Google Scholar 

  32. Pusch M, Neher E (1988) Rates of diffusional exchange between small cells and a measuring patch pipette. Pflügers Arch 411:204–211

    Google Scholar 

  33. Rane SG, Walsh MP, McDonald JR, Dunlap K (1989) Specific inhibitors of protein kinase C block transmitter-induced modulation of sensory neuron calcium current. Neuron 3:239–245

    Google Scholar 

  34. Rüegg UT, Burgess GM (1989) Staurosporine, K-252 and UCN-01: potent but nonspecific inhibitors of protein kinases. Trends Pharmacol Sci 10:218–220

    Google Scholar 

  35. Stanford IM, Chad JE (1991) Evaluation of voltage-and Ca++-dependent mechanisms in the inactivation of the Ca++ current in Helix neurons (abstract). Soc Neurosci Abstr 17:1517

    Google Scholar 

  36. Strong JA, Fox AP, Tsien RW, Kaczmarek LK (1987) Stimulation of protein kinase C recruits covert calcium channels in Aplysia bag cell neurons. Nature 325:714–717

    Google Scholar 

  37. Tiaho F, Richard S, Lory P, Nerbonne JM, Nargeot J (1990) Cyclic-AMP-dependent phosphorylation modulates the stereospecific activation of cardiac Ca channels by Bay K 8644. Pflügers Arch 417:58–66

    Google Scholar 

  38. Trautwein W, Hescheler J (1990) Regulation of cardiac L-type calcium current by phosphorylation and G-proteins. Annu Rev Physiol 52:257–274

    Google Scholar 

  39. Yakel JL (1991) The neuropeptide FMRFa both inhibits and enhances the Ca2+ current in dissociated Helix neurons via independent mechanisms. J Neurophysiol 65:1517–1527

    Google Scholar 

  40. Yount RG (1975) ATP analogues. Adv Enzymol 34:1–56

    Google Scholar 

Download references

Author information

Author notes

  1. Jerrel L. Yakel

    Present address: Vollum Institute, Oregon Health Sciences University, 3181 S. W. Sam Jackson Park Road, 97201, Portland, OR, USA

Authors and Affiliations

  1. Laboratoire de Neurobiologie (URA 295 CNRS), Ecole Normale Supérieure, 46, rue d'Ulm, F-75230, Paris Cedex 05, France

    Jerrel L. Yakel

Authors
  1. Jerrel L. Yakel
    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

Yakel, J.L. Inactivation of the Ba2+ current in dissociated Helix neurons: voltage dependence and the role of phosphorylation. Pflugers Arch. 420, 470–478 (1992). https://doi.org/10.1007/BF00374621

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00374621

Key words

Search

Navigation