Abstract
Some dyes of the methylene blue family serve as artificial inactivators of the sodium channels when present inside squid axons at a concentration of approximately 0.1 mM. The dyes restore a semblance of inactivation after normal inactivation has been destroyed by pronase. In fibers that inactivate normally, the dyes hasten the decay of sodium current. Many dye-blocked channels conduct transiently on exit of the dye molecule after repolarization to the holding potential. In contrast, normally inactivated channels do not conduct during recovery from inactivation. Kinetic evidence shows that inactivation of a dye- blocked channel is unlikely or impossible, which suggests that dye molecules compete with inactivation "particles" for the same site. In the absence of tetrodotoxin, the dyes do not affect the ON gating current unless the interpulse interval is very short. If sufficient equilibration time is allowed during a pulse, the initial amplitude of the OFF gating current is reduced to near zero. This suggests that a dye molecule is a Na channel completely blocks that channel's gating current, even the fraction that is resistant to normal inactivation. Dyes block INa and Ig with the same time course. This provides the strongest evidence to date that virtually all of recorded "gating current" is associated with Na channels. Tetrodotoxin greatly slows dissociation of dye molecules from Na channels and reduced gating current during both opening and closing of the channels.
Full Text
The Full Text of this article is available as a PDF (1.1 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Armstrong C. M., Bezanilla F. Inactivation of the sodium channel. II. Gating current experiments. J Gen Physiol. 1977 Nov;70(5):567–590. doi: 10.1085/jgp.70.5.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Armstrong C. M., Bezanilla F., Rojas E. Destruction of sodium conductance inactivation in squid axons perfused with pronase. J Gen Physiol. 1973 Oct;62(4):375–391. doi: 10.1085/jgp.62.4.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Armstrong C. M., Gilly W. F. Fast and slow steps in the activation of sodium channels. J Gen Physiol. 1979 Dec;74(6):691–711. doi: 10.1085/jgp.74.6.691. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bean B. P. Sodium channel inactivation in the crayfish giant axon. Must channels open before inactivating? Biophys J. 1981 Sep;35(3):595–614. doi: 10.1016/S0006-3495(81)84815-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bezanilla F., Armstrong C. M. Inactivation of the sodium channel. I. Sodium current experiments. J Gen Physiol. 1977 Nov;70(5):549–566. doi: 10.1085/jgp.70.5.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cahalan M. D., Almers W. Block of sodium conductance and gating current in squid giant axons poisoned with quaternary strychnine. Biophys J. 1979 Jul;27(1):57–73. doi: 10.1016/S0006-3495(79)85202-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cahalan M. D. Local anesthetic block of sodium channels in normal and pronase-treated squid giant axons. Biophys J. 1978 Aug;23(2):285–311. doi: 10.1016/S0006-3495(78)85449-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Courtney K. R. Mechanism of frequency-dependent inhibition of sodium currents in frog myelinated nerve by the lidocaine derivative GEA. J Pharmacol Exp Ther. 1975 Nov;195(2):225–236. [PubMed] [Google Scholar]
- Greeff N. G., Keynes R. D., Van Helden D. F. Fractionation of the asymmetry current in the squid giant axon into inactivating and non-inactivating components. Proc R Soc Lond B Biol Sci. 1982 Jun 22;215(1200):375–389. doi: 10.1098/rspb.1982.0048. [DOI] [PubMed] [Google Scholar]
- 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]
- Hille B. Local anesthetics: hydrophilic and hydrophobic pathways for the drug-receptor reaction. J Gen Physiol. 1977 Apr;69(4):497–515. doi: 10.1085/jgp.69.4.497. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kirsch G. E., Yeh J. Z., Farley J. M., Narahashi T. Interaction of n-alkylguanidines with the sodium channels of squid axon membrane. J Gen Physiol. 1980 Sep;76(3):315–335. doi: 10.1085/jgp.76.3.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Meves H., Vogel W. Inactivation of the asymmetrical displacement current in giant axons of Loligo forbesi. J Physiol. 1977 May;267(2):377–393. doi: 10.1113/jphysiol.1977.sp011818. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nonner W. Relations between the inactivation of sodium channels and the immobilization of gating charge in frog myelinated nerve. J Physiol. 1980 Feb;299:573–603. doi: 10.1113/jphysiol.1980.sp013143. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Oxford G. S., Pooler J. P., Narahashi T. Internal and external application of photodynamic sensitizers on squid giant axons. J Membr Biol. 1977 Sep 14;36(2-3):159–173. doi: 10.1007/BF01868149. [DOI] [PubMed] [Google Scholar]
- Oxford G. S., Wu C. H., Narahashi T. Removal of sodium channel inactivation in squid giant axons by n-bromoacetamide. J Gen Physiol. 1978 Mar;71(3):227–247. doi: 10.1085/jgp.71.3.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pooler J. Photodynamic alteration of sodium currents in lobster axons. J Gen Physiol. 1972 Oct;60(4):367–387. doi: 10.1085/jgp.60.4.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rojas E., Rudy B. Destruction of the sodium conductance inactivation by a specific protease in perfused nerve fibres from Loligo. J Physiol. 1976 Nov;262(2):501–531. doi: 10.1113/jphysiol.1976.sp011608. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shapiro B. I. Effects of strychnine on the sodium conductance of the frog node of Ranvier. J Gen Physiol. 1977 Jun;69(6):915–926. doi: 10.1085/jgp.69.6.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yeh J. Z., Armstrong C. M. Immobilisation of gating charge by a substance that simulates inactivation. Nature. 1978 Jun 1;273(5661):387–389. doi: 10.1038/273387a0. [DOI] [PubMed] [Google Scholar]
- Yeh J. Z., Narahashi T. Kinetic analysis of pancuronium interaction with sodium channels in squid axon membranes. J Gen Physiol. 1977 Mar;69(3):293–323. doi: 10.1085/jgp.69.3.293. [DOI] [PMC free article] [PubMed] [Google Scholar]