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. 1993 Sep;65(3):1196–1206. doi: 10.1016/S0006-3495(93)81153-6

Slow and incomplete inactivations of voltage-gated channels dominate encoding in synthetic neurons.

H Hsu 1, E Huang 1, X C Yang 1, A Karschin 1, C Labarca 1, A Figl 1, B Ho 1, N Davidson 1, H A Lester 1
PMCID: PMC1225839  PMID: 8241400

Abstract

Electrically excitable channels were expressed in Chinese hamster ovary cells using a vaccinia virus vector system. In cells expressing rat brain IIA Na+ channels only, brief pulses (< 1 ms) of depolarizing current resulted in action potentials with a prolonged (0.5-3 s) depolarizing plateau; this plateau was caused by slow and incomplete Na+ channel inactivation. In cells expressing both Na+ and Drosophila Shaker H4 transient K+ channels, there were neuron-like action potentials. In cells with appropriate Na+/K+ current ratios, maintaining stimulation produced repetitive firing over a 10-fold range of frequencies but eventually led to "lock-up" of the potential at a positive value after several seconds of stimulation. The latter effect was due primarily to slow inactivation of the K+ currents. Numerical simulations of modified Hodgkin-Huxley equations describing these currents, using parameters from voltage-clamp kinetics studied in the same cells, accounted for most features of the voltage trajectories. The present study shows that insights into the mechanisms for generating action potentials and trains of action potentials in real excitable cells can be obtained from the analysis of synthetic excitable cells that express a controlled repertoire of ion channels.

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Selected References

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

  1. Andersen P., Raastad M., Storm J. F. Excitatory synaptic integration in hippocampal pyramids and dentate granule cells. Cold Spring Harb Symp Quant Biol. 1990;55:81–86. doi: 10.1101/sqb.1990.055.01.010. [DOI] [PubMed] [Google Scholar]
  2. Augustine G. J. Regulation of transmitter release at the squid giant synapse by presynaptic delayed rectifier potassium current. J Physiol. 1990 Dec;431:343–364. doi: 10.1113/jphysiol.1990.sp018333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Auld V. J., Goldin A. L., Krafte D. S., Catterall W. A., Lester H. A., Davidson N., Dunn R. J. A neutral amino acid change in segment IIS4 dramatically alters the gating properties of the voltage-dependent sodium channel. Proc Natl Acad Sci U S A. 1990 Jan;87(1):323–327. doi: 10.1073/pnas.87.1.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Auld V. J., Goldin A. L., Krafte D. S., Marshall J., Dunn J. M., Catterall W. A., Lester H. A., Davidson N., Dunn R. J. A rat brain Na+ channel alpha subunit with novel gating properties. Neuron. 1988 Aug;1(6):449–461. doi: 10.1016/0896-6273(88)90176-6. [DOI] [PubMed] [Google Scholar]
  5. Belluzzi O., Sacchi O., Wanke E. Identification of delayed potassium and calcium currents in the rat sympathetic neurone under voltage clamp. J Physiol. 1985 Jan;358:109–129. doi: 10.1113/jphysiol.1985.sp015543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Blanton M. G., Lo Turco J. J., Kriegstein A. R. Whole cell recording from neurons in slices of reptilian and mammalian cerebral cortex. J Neurosci Methods. 1989 Dec;30(3):203–210. doi: 10.1016/0165-0270(89)90131-3. [DOI] [PubMed] [Google Scholar]
  7. Chandler W. K., Meves H. Evidence for two types of sodium conductance in axons perfused with sodium fluoride solution. J Physiol. 1970 Dec;211(3):653–678. doi: 10.1113/jphysiol.1970.sp009298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chandler W. K., Meves H. Rate constants associated with changes in sodium conductance in axons perfused with sodium fluoride. J Physiol. 1970 Dec;211(3):679–705. doi: 10.1113/jphysiol.1970.sp009299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chandler W. K., Meves H. Slow changes in membrane permeability and long-lasting action potentials in axons perfused with fluoride solutions. J Physiol. 1970 Dec;211(3):707–728. doi: 10.1113/jphysiol.1970.sp009300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chandler W. K., Meves H. Sodium and potassium currents in squid axons perfused with fluoride solutions. J Physiol. 1970 Dec;211(3):623–652. doi: 10.1113/jphysiol.1970.sp009297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Connor J. A., Stevens C. F. Inward and delayed outward membrane currents in isolated neural somata under voltage clamp. J Physiol. 1971 Feb;213(1):1–19. doi: 10.1113/jphysiol.1971.sp009364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Connor J. A., Stevens C. F. Prediction of repetitive firing behaviour from voltage clamp data on an isolated neurone soma. J Physiol. 1971 Feb;213(1):31–53. doi: 10.1113/jphysiol.1971.sp009366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Edwards F. A., Konnerth A., Sakmann B., Takahashi T. A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system. Pflugers Arch. 1989 Sep;414(5):600–612. doi: 10.1007/BF00580998. [DOI] [PubMed] [Google Scholar]
  15. Elroy-Stein O., Fuerst T. R., Moss B. Cap-independent translation of mRNA conferred by encephalomyocarditis virus 5' sequence improves the performance of the vaccinia virus/bacteriophage T7 hybrid expression system. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6126–6130. doi: 10.1073/pnas.86.16.6126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Grant A. O., Starmer C. F. Mechanisms of closure of cardiac sodium channels in rabbit ventricular myocytes: single-channel analysis. Circ Res. 1987 Jun;60(6):897–913. doi: 10.1161/01.res.60.6.897. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. HUXLEY A. F. Ion movements during nerve activity. Ann N Y Acad Sci. 1959 Aug 28;81:221–246. doi: 10.1111/j.1749-6632.1959.tb49311.x. [DOI] [PubMed] [Google Scholar]
  19. Hines M. A program for simulation of nerve equations with branching geometries. Int J Biomed Comput. 1989 Mar;24(1):55–68. doi: 10.1016/0020-7101(89)90007-x. [DOI] [PubMed] [Google Scholar]
  20. Hoshi T., Zagotta W. N., Aldrich R. W. Two types of inactivation in Shaker K+ channels: effects of alterations in the carboxy-terminal region. Neuron. 1991 Oct;7(4):547–556. doi: 10.1016/0896-6273(91)90367-9. [DOI] [PubMed] [Google Scholar]
  21. Iverson L. E., Rudy B. The role of the divergent amino and carboxyl domains on the inactivation properties of potassium channels derived from the Shaker gene of Drosophila. J Neurosci. 1990 Sep;10(9):2903–2916. doi: 10.1523/JNEUROSCI.10-09-02903.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Iverson L. E., Tanouye M. A., Lester H. A., Davidson N., Rudy B. A-type potassium channels expressed from Shaker locus cDNA. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5723–5727. doi: 10.1073/pnas.85.15.5723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kaang B. K., Pfaffinger P. J., Grant S. G., Kandel E. R., Furukawa Y. Overexpression of an Aplysia shaker K+ channel gene modifies the electrical properties and synaptic efficacy of identified Aplysia neurons. Proc Natl Acad Sci U S A. 1992 Feb 1;89(3):1133–1137. doi: 10.1073/pnas.89.3.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kamb A., Iverson L. E., Tanouye M. A. Molecular characterization of Shaker, a Drosophila gene that encodes a potassium channel. Cell. 1987 Jul 31;50(3):405–413. doi: 10.1016/0092-8674(87)90494-6. [DOI] [PubMed] [Google Scholar]
  25. Karschin A., Aiyar J., Gouin A., Davidson N., Lester H. A. K+ channel expression in primary cell cultures mediated by vaccinia virus. FEBS Lett. 1991 Jan 28;278(2):229–233. doi: 10.1016/0014-5793(91)80123-k. [DOI] [PubMed] [Google Scholar]
  26. Karschin A., Ho B. Y., Labarca C., Elroy-Stein O., Moss B., Davidson N., Lester H. A. Heterologously expressed serotonin 1A receptors couple to muscarinic K+ channels in heart. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5694–5698. doi: 10.1073/pnas.88.13.5694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Klaiber K., Williams N., Roberts T. M., Papazian D. M., Jan L. Y., Miller C. Functional expression of Shaker K+ channels in a baculovirus-infected insect cell line. Neuron. 1990 Aug;5(2):221–226. doi: 10.1016/0896-6273(90)90311-3. [DOI] [PubMed] [Google Scholar]
  28. Klein M., Camardo J., Kandel E. R. Serotonin modulates a specific potassium current in the sensory neurons that show presynaptic facilitation in Aplysia. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5713–5717. doi: 10.1073/pnas.79.18.5713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Krafte D. S., Goldin A. L., Auld V. J., Dunn R. J., Davidson N., Lester H. A. Inactivation of cloned Na channels expressed in Xenopus oocytes. J Gen Physiol. 1990 Oct;96(4):689–706. doi: 10.1085/jgp.96.4.689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Krafte D. S., Snutch T. P., Leonard J. P., Davidson N., Lester H. A. Evidence for the involvement of more than one mRNA species in controlling the inactivation process of rat and rabbit brain Na channels expressed in Xenopus oocytes. J Neurosci. 1988 Aug;8(8):2859–2868. doi: 10.1523/JNEUROSCI.08-08-02859.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Leonard R. J., Garcia M. L., Slaughter R. S., Reuben J. P. Selective blockers of voltage-gated K+ channels depolarize human T lymphocytes: mechanism of the antiproliferative effect of charybdotoxin. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10094–10098. doi: 10.1073/pnas.89.21.10094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Leonard R. J., Karschin A., Jayashree-Aiyar S., Davidson N., Tanouye M. A., Thomas L., Thomas G., Lester H. A. Expression of Drosophila Shaker potassium channels in mammalian cells infected with recombinant vaccinia virus. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7629–7633. doi: 10.1073/pnas.86.19.7629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Liu Y. M., DeFelice L. J., Mazzanti M. Na channels that remain open throughout the cardiac action potential plateau. Biophys J. 1992 Sep;63(3):654–662. doi: 10.1016/S0006-3495(92)81635-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Llinás R. R. The intrinsic electrophysiological properties of mammalian neurons: insights into central nervous system function. Science. 1988 Dec 23;242(4886):1654–1664. doi: 10.1126/science.3059497. [DOI] [PubMed] [Google Scholar]
  35. Marom S., Goldstein S. A., Kupper J., Levitan I. B. Mechanism and modulation of inactivation of the Kv3 potassium channel. Receptors Channels. 1993;1(1):81–88. [PubMed] [Google Scholar]
  36. Matteson D. R., Armstrong C. M. Evidence for a population of sleepy sodium channels in squid axon at low temperature. J Gen Physiol. 1982 May;79(5):739–758. doi: 10.1085/jgp.79.5.739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Moorman J. R., Kirsch G. E., VanDongen A. M., Joho R. H., Brown A. M. Fast and slow gating of sodium channels encoded by a single mRNA. Neuron. 1990 Feb;4(2):243–252. doi: 10.1016/0896-6273(90)90099-2. [DOI] [PubMed] [Google Scholar]
  38. Moss B., Elroy-Stein O., Mizukami T., Alexander W. A., Fuerst T. R. Product review. New mammalian expression vectors. Nature. 1990 Nov 1;348(6296):91–92. doi: 10.1038/348091a0. [DOI] [PubMed] [Google Scholar]
  39. Patlak J. B., Ortiz M. Slow currents through single sodium channels of the adult rat heart. J Gen Physiol. 1985 Jul;86(1):89–104. doi: 10.1085/jgp.86.1.89. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Patlak J. B., Ortiz M. Two modes of gating during late Na+ channel currents in frog sartorius muscle. J Gen Physiol. 1986 Feb;87(2):305–326. doi: 10.1085/jgp.87.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Rudy B. Diversity and ubiquity of K channels. Neuroscience. 1988 Jun;25(3):729–749. doi: 10.1016/0306-4522(88)90033-4. [DOI] [PubMed] [Google Scholar]
  42. Rudy B. Slow inactivation of the sodium conductance in squid giant axons. Pronase resistance. J Physiol. 1978 Oct;283:1–21. doi: 10.1113/jphysiol.1978.sp012485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Timpe L. C., Schwarz T. L., Tempel B. L., Papazian D. M., Jan Y. N., Jan L. Y. Expression of functional potassium channels from Shaker cDNA in Xenopus oocytes. Nature. 1988 Jan 14;331(6152):143–145. doi: 10.1038/331143a0. [DOI] [PubMed] [Google Scholar]
  44. Yang X. C., Labarca C., Nargeot J., Ho B. Y., Elroy-Stein O., Moss B., Davidson N., Lester H. A. Cell-specific posttranslational events affect functional expression at the plasma membrane but not tetrodotoxin sensitivity of the rat brain IIA sodium channel alpha-subunit expressed in mammalian cells. J Neurosci. 1992 Jan;12(1):268–277. doi: 10.1523/JNEUROSCI.12-01-00268.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Zhou J. Y., Potts J. F., Trimmer J. S., Agnew W. S., Sigworth F. J. Multiple gating modes and the effect of modulating factors on the microI sodium channel. Neuron. 1991 Nov;7(5):775–785. doi: 10.1016/0896-6273(91)90280-d. [DOI] [PubMed] [Google Scholar]

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