Skip to main content
The Journal of Physiology logoLink to The Journal of Physiology
. 1978 May;278:265–286. doi: 10.1113/jphysiol.1978.sp012303

Ionic currents in cultured mouse neuroblastoma cells under voltage-clamp conditions.

W H Moolenaar, I Spector
PMCID: PMC1282348  PMID: 671297

Abstract

1. Ionic currents in differentiated cells of mouse neuroblastoma clone N1E-115 have been studied under voltage-clamp conditions. 2. Depolarizing voltage steps from a holding potential of -85 mV to levels more positive than -40 mV produced fast transient inward currents followed by delayed outward currents. 3. The fast inward current is carried by Na+: it is blocked by tetrodotoxin and is absent in Na+-free solutions. Its kinetic behaviour resembles that of the Na+ current in squid giant axon. A mean value of 85 mmho/cm2 was found for the maximum Na+ conductance (GNa).4. The delayed outward current is carried primarily by K+: it is blocked by externally applied tetraethylammonium (TEA, 15 mM) and has a reversal potential (mean -71 mV) close to the theoretical K+ equilibrium potential. Its instantaneous I--V curve is linear. By analogy with the formulation of Hodgkin & Huxley (1952c), the outward current can be described by IK = -GKn2(V--EK) where GK = 12 mmho/mc2. 5. During prolonged depolarizations the delayed outward current declines. This decline, which occurs in two phases, represents a partial inactivation of the K+ conductance. 6. A weak inward current with slow activation and inactivation kinetics appears in Na+-free solution containing 10 mM-Ca2+. It is activated at a membrane potential of -55 mV and reaches its maximum at -20 mV with a time to peak of about 10 msec. This current is tetrodotoxin-resistant, reversibly blocked by Co2+ (5mM) and is suggested to be carried by Ca2+. 7. An increase in the external divalent cation concentration results in a parallel shift of the steady-state I--V curve along the voltage axis in positive direction. The activation of delayed outward currents is suggested not to depend on Ca2+ influx. 8. It is concluded that separate voltage-dependent Na+, K+ and Ca2+ channels exist in the differentiated neuroblastoma membrane with kinetic and pharmacological properties similar to those observed in non-mammalian preparations.

Full text

PDF
265

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ARAKI T., TERZUOLO C. A. Membrane currents in spinal motoneurons associated with the action potential and synaptic activity. J Neurophysiol. 1962 Nov;25:772–789. doi: 10.1152/jn.1962.25.6.772. [DOI] [PubMed] [Google Scholar]
  2. Amano T., Richelson E., Nirenberg M. Neurotransmitter synthesis by neuroblastoma clones (neuroblast differentiation-cell culture-choline acetyltransferase-acetylcholinesterase-tyrosine hydroxylase-axons-dendrites). Proc Natl Acad Sci U S A. 1972 Jan;69(1):258–263. doi: 10.1073/pnas.69.1.258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baccaglini P. I., Spitzer N. C. Developmental changes in the inward current of the action potential of Rohon-Beard neurones. J Physiol. 1977 Sep;271(1):93–117. doi: 10.1113/jphysiol.1977.sp011992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baker P. F., Hodgkin A. L., Ridgway E. B. Depolarization and calcium entry in squid giant axons. J Physiol. 1971 Nov;218(3):709–755. doi: 10.1113/jphysiol.1971.sp009641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barrett E. F., Barret J. N. Separation of two voltage-sensitive potassium currents, and demonstration of a tetrodotoxin-resistant calcium current in frog motoneurones. J Physiol. 1976 Mar;255(3):737–774. doi: 10.1113/jphysiol.1976.sp011306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Begenisich T., Lynch C. Effects of internal divalent cations on voltage-clamped squid axons. J Gen Physiol. 1974 Jun;63(6):675–689. doi: 10.1085/jgp.63.6.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Connor J. A., Stevens C. F. Voltage clamp studies of a transient outward membrane current in gastropod neural somata. J Physiol. 1971 Feb;213(1):21–30. doi: 10.1113/jphysiol.1971.sp009365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dichter M. A., Fischbach G. D. The action potential of chick dorsal root ganglion neurones maintained in cell culture. J Physiol. 1977 May;267(2):281–298. doi: 10.1113/jphysiol.1977.sp011813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eckert R., Lux H. D. A voltage-sensitive persistent calcium conductance in neuronal somata of Helix. J Physiol. 1976 Jan;254(1):129–151. doi: 10.1113/jphysiol.1976.sp011225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. FRANKENHAEUSER B. A QUANTITATIVE DESCRIPTION OF POTASSIUM CURRENTS IN MYELINATED NERVE FIBRES OF XENOPUS LAEVIS. J Physiol. 1963 Nov;169:424–430. doi: 10.1113/jphysiol.1963.sp007268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. FRANKENHAEUSER B., HODGKIN A. L. The action of calcium on the electrical properties of squid axons. J Physiol. 1957 Jul 11;137(2):218–244. doi: 10.1113/jphysiol.1957.sp005808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. FRANK K., FUORTES M. G., NELSON P. G. Voltage clamp of motoneuron soma. Science. 1959 Jul 3;130(3366):38–39. doi: 10.1126/science.130.3366.38. [DOI] [PubMed] [Google Scholar]
  13. Gilbert D. L., Ehrenstein G. Effect of divalent cations on potassium conductance of squid axons: determination of surface charge. Biophys J. 1969 Mar;9(3):447–463. doi: 10.1016/S0006-3495(69)86396-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Goldman L., Schauf C. L. Quantitative description of sodium and potassium currents and computed action potentials in Myxicola giant axons. J Gen Physiol. 1973 Mar;61(3):361–384. doi: 10.1085/jgp.61.3.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. HAGIWARA S., SAITO N. Voltage-current relations in nerve cell membrane of Onchidium verruculatum. J Physiol. 1959 Oct;148:161–179. doi: 10.1113/jphysiol.1959.sp006279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. HODGKIN A. L., HUXLEY A. F. A quantitative description of membrane current and its application to conduction and excitation in nerve. J Physiol. 1952 Aug;117(4):500–544. doi: 10.1113/jphysiol.1952.sp004764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. HODGKIN A. L., HUXLEY A. F. Currents carried by sodium and potassium ions through the membrane of the giant axon of Loligo. J Physiol. 1952 Apr;116(4):449–472. doi: 10.1113/jphysiol.1952.sp004717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. HODGKIN A. L., HUXLEY A. F. The dual effect of membrane potential on sodium conductance in the giant axon of Loligo. J Physiol. 1952 Apr;116(4):497–506. doi: 10.1113/jphysiol.1952.sp004719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hagiwara S., Takahashi K. Surface density of calcium ions and calcium spikes in the barnacle muscle fiber membrane. J Gen Physiol. 1967 Jan;50(3):583–601. doi: 10.1085/jgp.50.3.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Heyer C. B., Lux H. D. Control of the delayed outward potassium currents in bursting pace-maker neurones of the snail, Helix pomatia. J Physiol. 1976 Nov;262(2):349–382. doi: 10.1113/jphysiol.1976.sp011599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hille B. Charges and potentials at the nerve surface. Divalent ions and pH. J Gen Physiol. 1968 Feb;51(2):221–236. doi: 10.1085/jgp.51.2.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kidokoro Y. Sodium and calcium components of the action potential in a developing skeletal muscle cell line. J Physiol. 1975 Jan;244(1):145–159. doi: 10.1113/jphysiol.1975.sp010788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kimhi Y., Palfrey C., Spector I., Barak Y., Littauer U. Z. Maturation of neuroblastoma cells in the presence of dimethylsulfoxide. Proc Natl Acad Sci U S A. 1976 Feb;73(2):462–466. doi: 10.1073/pnas.73.2.462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kleinhaus A. L., Prichard J. W. Calcium dependent action potentials produced in leech Retzius cells by tetraethylammonium chloride. J Physiol. 1975 Mar;246(2):351–369. doi: 10.1113/jphysiol.1975.sp010894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Koketsu K., Nishi S. Calcium and action potentials of bullfrog sympathetic ganglion cells. J Gen Physiol. 1969 May;53(5):608–623. doi: 10.1085/jgp.53.5.608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Llinás R., Steinberg I. Z., Walton K. Presynaptic calcium currents and their relation to synaptic transmission: voltage clamp study in squid giant synapse and theoretical model for the calcium gate. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2918–2922. doi: 10.1073/pnas.73.8.2918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Matsuda Y., Yoshida S., Yonezawa T. A Ca- dependent regenerative response in rodent dorsal root ganglion cells cultured in vitro. Brain Res. 1976 Oct 15;115(2):334–338. doi: 10.1016/0006-8993(76)90519-9. [DOI] [PubMed] [Google Scholar]
  28. Meech R. W., Standen N. B. Potassium activation in Helix aspersa neurones under voltage clamp: a component mediated by calcium influx. J Physiol. 1975 Jul;249(2):211–239. doi: 10.1113/jphysiol.1975.sp011012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Moolenaar W. H., Spector I. Membrane currents examined under voltage clamp in cultured neuroblastoma cells. Science. 1977 Apr 15;196(4287):331–333. doi: 10.1126/science.557842. [DOI] [PubMed] [Google Scholar]
  30. NARAHASHI T., MOORE J. W., SCOTT W. R. TETRODOTOXIN BLOCKAGE OF SODIUM CONDUCTANCE INCREASE IN LOBSTER GIANT AXONS. J Gen Physiol. 1964 May;47:965–974. doi: 10.1085/jgp.47.5.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Narahashi T. Chemicals as tools in the study of excitable membranes. Physiol Rev. 1974 Oct;54(4):813–889. doi: 10.1152/physrev.1974.54.4.813. [DOI] [PubMed] [Google Scholar]
  32. Neher E. Two fast transient current components during voltage clamp on snail neurons. J Gen Physiol. 1971 Jul;58(1):36–53. doi: 10.1085/jgp.58.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nelson P. G. Nerve and muscle cells in culture. Physiol Rev. 1975 Jan;55(1):1–61. doi: 10.1152/physrev.1975.55.1.1. [DOI] [PubMed] [Google Scholar]
  34. Okamoto H., Takahashi K., Yamashita N. Ionic currents through the membrane of the mammalian oocyte and their comparison with those in the tunicate and sea urchin. J Physiol. 1977 May;267(2):465–495. doi: 10.1113/jphysiol.1977.sp011822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Okamoto H., Takahashi K., Yoshii M. Membrane currents of the tunicate egg under the voltage-clamp condition. J Physiol. 1976 Jan;254(3):607–638. doi: 10.1113/jphysiol.1976.sp011249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Prasad K. N. Differentiation of neuroblastoma cells in culture. Biol Rev Camb Philos Soc. 1975 May;50(2):129–165. doi: 10.1111/j.1469-185x.1975.tb01055.x. [DOI] [PubMed] [Google Scholar]
  37. Reuter H. Divalent cations as charge carriers in excitable membranes. Prog Biophys Mol Biol. 1973;26:1–43. doi: 10.1016/0079-6107(73)90016-3. [DOI] [PubMed] [Google Scholar]
  38. Schwarz J. R., Vogel W. Potassium inactivation in single myelinated nerve fibres of Xenopus laevis. Pflugers Arch. 1971;330(1):61–73. doi: 10.1007/BF00588735. [DOI] [PubMed] [Google Scholar]
  39. Schwindt P., Crill W. E. A persistent negative resistance in cat lumbar motoneurons. Brain Res. 1977 Jan 14;120(1):173–178. doi: 10.1016/0006-8993(77)90510-8. [DOI] [PubMed] [Google Scholar]
  40. Spector I., Kimhi Y., Nelson P. G. Tetrodotoxin and cobalt blockade of neuroblastoma action potentials. Nat New Biol. 1973 Nov 28;246(152):124–126. doi: 10.1038/newbio246124a0. [DOI] [PubMed] [Google Scholar]
  41. Standen N. B. Voltage-clamp studies of the calcium inward current in an identified snail neurone: comparison with the sodium inward current. J Physiol. 1975 Jul;249(2):253–268. doi: 10.1113/jphysiol.1975.sp011014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. TASAKI I., HAGIWAR A. S. Demonstration of two stable potential states in the squid giant axon under tetraethylammonium chloride. J Gen Physiol. 1957 Jul 20;40(6):859–885. doi: 10.1085/jgp.40.6.859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Thompson S. H. Three pharmacologically distinct potassium channels in molluscan neurones. J Physiol. 1977 Feb;265(2):465–488. doi: 10.1113/jphysiol.1977.sp011725. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES