Home LiteratureArticle Details
PMID: 2435540 Published · ppublish English Journal Article

Patch clamp characterization of sodium channels expressed from rat brain cDNA.

European biophysics journal : EBJ ·Vol. 14 ·No. 3 ·1987-00-00 ·Pages 131-8

Stühmer W, Methfessel C, Sakmann B, Noda M, Numa S

Abstract

Sodium currents, INa, were recorded from Xenopus laevis oocytes which had been injected with mRNA synthesized by in vitro transcription of the rat brain sodium channel II cDNA (Noda et al. 1986 a, b). Patch pipettes were used to apply depolarizing voltage steps and to record macroscopic sodium currents of between 50 and 750 pA from cell-attached patches of the oocyte membrane. With a combination of whole-cell and patch clamp recording, the properties of the implanted sodium channels could be studied in detail. They were analyzed according to the model of Hodgkin and Huxley (1952 a) assuming three activation gates. The activation of the sodium currents is characterized by an equilibrium potential of -29 mV and an apparent gating charge of 8.7 e0. At -64 mV half of the sodium currents were inactivated. From single-channel current recordings, an elementary sodium channel conductance of 19 pS and an average open time of 0.43 ms were obtained at -32 mV membrane potential and 16 degrees C. The single-channel and activation properties of rat brain sodium channel II are therefore comparable to those found in peripheral nerve and skeletal muscle, but inactivation occurs at less negative potentials. This could be a specific property of the brain sodium channels and may underlie the maintained inward sodium currents reported in brain neurones (French and Gage 1985).

MeSH Terms
Animals Brain/physiology DNA/metabolism Female Ion Channels/physiology Membrane Potentials Membrane Proteins/genetics Oocytes/metabolism RNA, Messenger/genetics Rats Sodium/metabolism Sodium Channels Transcription, Genetic Xenopus
Chemicals
Ion Channels Membrane Proteins RNA, Messenger Sodium Channels DNA Sodium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Stühmer W
Methfessel C
Sakmann B
Noda M
Numa S
References (26)
26 references, click to expand
  1. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.
    Pflugers Arch. 1981 Aug;391(2):85-100 PMID: 6270629
  2. Patch clamp measurements on Xenopus laevis oocytes: currents through endogenous channels and implanted acetylcholine receptor and sodium channels.
    Pflugers Arch. 1986 Dec;407(6):577-88 PMID: 2432468
  3. Sodium currents and sodium-current fluctuations in rat myelinated nerve fibres.
    J Physiol. 1982 Aug;329:163-84 PMID: 6292404
  4. Comparison between slow sodium channel inactivation in rat slow- and fast-twitch muscle.
    J Physiol. 1987 Feb;383:339-48 PMID: 2443650
  5. Slow actions of hyperpolarization on sodium channels in the membrane of myelinated nerve.
    Biochim Biophys Acta. 1979 Nov 16;558(1):113-8 PMID: 315238
  6. Single Na+ channel currents observed in cultured rat muscle cells.
    Nature. 1980 Oct 2;287(5781):447-9 PMID: 6253802
  7. Messenger RNA from human brain induces drug- and voltage-operated channels in Xenopus oocytes.
    Nature. 1984 Mar 29-Apr 4;308(5958):421-4 PMID: 6323990
  8. A threshold sodium current in pyramidal cells in rat hippocampus.
    Neurosci Lett. 1985 May 23;56(3):289-93 PMID: 2410817
  9. Role of acetylcholine receptor subunits in gating of the channel.
    Nature. 1985 Dec 12-18;318(6046):538-43 PMID: 2415826
  10. Existence of distinct sodium channel messenger RNAs in rat brain.
    Nature. 1986 Mar 13-19;320(6058):188-92 PMID: 3754035
  11. Slow sodium channel inactivation in rat fast-twitch muscle.
    J Physiol. 1987 Feb;383:327-37 PMID: 2443649
  12. Expression of functional sodium channels from cloned cDNA.
    Nature. 1986 Aug 28-Sep 3;322(6082):826-8 PMID: 2427955
  13. Partial purification and functional expression of brain mRNAs coding for neurotransmitter receptors and voltage-operated channels.
    Proc Natl Acad Sci U S A. 1984 Dec;81(24):7994-8 PMID: 6151179
  14. Sodium channels induced by depolarization of the Xenopus laevis oocyte.
    Proc Natl Acad Sci U S A. 1982 May;79(10):3188-92 PMID: 6285341
  15. Voltage-operated channels induced by foreign messenger RNA in Xenopus oocytes.
    Proc R Soc Lond B Biol Sci. 1983 Nov 22;220(1218):131-40 PMID: 6140681
  16. Quantitative description of sodium currents in myelinated nerve fibres of Xenopus laevis.
    J Physiol. 1960 Jun;151:491-501 PMID: 13824558
  17. Characterization of mRNA responsible for induction of functional sodium channels in Xenopus oocytes.
    Brain Res. 1985 Dec 16;359(1-2):57-64 PMID: 2416399
  18. The variance of sodium current fluctuations at the node of Ranvier.
    J Physiol. 1980 Oct;307:97-129 PMID: 6259340
  19. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
    Nucleic Acids Res. 1984 Sep 25;12(18):7035-56 PMID: 6091052
  20. Human beta-globin pre-mRNA synthesized in vitro is accurately spliced in Xenopus oocyte nuclei.
    Cell. 1983 Mar;32(3):681-94 PMID: 6550524
  21. Molecular distinction between fetal and adult forms of muscle acetylcholine receptor.
    Nature. 1986 May 22-28;321(6068):406-11 PMID: 2423878
  22. The dual effect of membrane potential on sodium conductance in the giant axon of Loligo.
    J Physiol. 1952 Apr;116(4):497-506 PMID: 14946715
  23. A quantitative description of membrane current and its application to conduction and excitation in nerve.
    J Physiol. 1952 Aug;117(4):500-44 PMID: 12991237
  24. Primary structure of Electrophorus electricus sodium channel deduced from cDNA sequence.
    Nature. 1984 Nov 8-14;312(5990):121-7 PMID: 6209577
  25. Voltage-clamp experiments in normal and denervated mammalian skeletal muscle fibres.
    J Physiol. 1980 Sep;306:377-410 PMID: 6257898
  26. Voltage clamp of rat and human skeletal muscle: measurements with an improved loose-patch technique.
    J Physiol. 1984 Feb;347:751-68 PMID: 6323705
Article Info
Journal
European biophysics journal : EBJ
Abbr.
Eur Biophys J
ISSN
0175-7571
Published
1987-00-00
Pages
131-8
Language
English
Region
Germany
NLM ID
8409413
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com