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. 1976 Mar 1;67(3):265–293. doi: 10.1085/jgp.67.3.265

An improved vaseline gap voltage clamp for skeletal muscle fibers

PMCID: PMC2214972  PMID: 1083424

Abstract

A Vaseline gap potentiometric recording and voltage clamp method is developed for frog skeletal muscle fibers. The method is based on the Frankenhaeuser-Dodge voltage clamp for myelinated nerve with modifications to improve the frequency response, to compensate for external series resistance, and to compensate for the complex impedance of the current-passing pathway. Fragments of single muscle fibers are plucked from the semitendinosus muscle and mounted while depolarized by a solution like CsF. After Vaseline seals are formed between fluid pools, the fiber ends are cut once again, the central region is rinsed with Ringer solution, and the feedback amplifiers are turned on. Errors in the potential and current records are assessed by direct measurements with microelectrodes. The passive properties of the preparation are simulated by the "disk" equivalent circuit for the transverse tubular system and the derived parameters are similar to previous measurements with microelectrodes. Action potentials at 5 degrees C are long because of the absence of delayed rectification. Their shape is approximately simulated by solving the disk model with sodium permeability in the surface and tubular membranes. Voltage clamp currents consist primarily of capacity currents and sodium currents. The peak inward sodium current density at 5 degrees C is 3.7 mA/cm2. At 5 degrees C the sodium currents are smoothly graded with increasing depolarization and free of notches suggesting good control of the surface membrane. At higher temperatures a small, late extra inward current appears for small depolarizations that has the properties expected for excitation in the transverse tubular system. Comparison of recorded currents with simulations shows that while the transverse tubular system has regenerative sodium currents, they are too small to make important errors in the total current recorded at the surface under voltage clamp at low temperature. The tubules are definitely not under voltage clamp control.

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Selected References

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  1. Adrian R. H., Almers W. Membrane capacity measurements on frog skeletal muscle in media of low ion content. J Physiol. 1974 Mar;237(3):573–605. doi: 10.1113/jphysiol.1974.sp010499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adrian R. H., Chandler W. K., Hodgkin A. L. Slow changes in potassium permeability in skeletal muscle. J Physiol. 1970 Jul;208(3):645–668. doi: 10.1113/jphysiol.1970.sp009140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Adrian R. H., Chandler W. K., Hodgkin A. L. The kinetics of mechanical activation in frog muscle. J Physiol. 1969 Sep;204(1):207–230. doi: 10.1113/jphysiol.1969.sp008909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Adrian R. H., Chandler W. K., Hodgkin A. L. Voltage clamp experiments in striated muscle fibres. J Physiol. 1970 Jul;208(3):607–644. doi: 10.1113/jphysiol.1970.sp009139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Adrian R. H., Peachey L. D. Reconstruction of the action potential of frog sartorius muscle. J Physiol. 1973 Nov;235(1):103–131. doi: 10.1113/jphysiol.1973.sp010380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Armstrong C. M., Bezanilla F. Charge movement associated with the opening and closing of the activation gates of the Na channels. J Gen Physiol. 1974 May;63(5):533–552. doi: 10.1085/jgp.63.5.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Armstrong C. M., Hille B. The inner quaternary ammonium ion receptor in potassium channels of the node of Ranvier. J Gen Physiol. 1972 Apr;59(4):388–400. doi: 10.1085/jgp.59.4.388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bastian J., Nakajima S. Action potential in the transverse tubules and its role in the activation of skeletal muscle. J Gen Physiol. 1974 Feb;63(2):257–278. doi: 10.1085/jgp.63.2.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Campbell D. T., Hille B. Kinetic and pharmacological properties of the sodium channel of frog skeletal muscle. J Gen Physiol. 1976 Mar;67(3):309–323. doi: 10.1085/jgp.67.3.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Campbell D. T. Ionic selectivity of the sodium channel of frog skeletal muscle. J Gen Physiol. 1976 Mar;67(3):295–307. doi: 10.1085/jgp.67.3.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Costantin L. L. The effect o f calcium on contraction and conductance thresholds in frog skeletal muscle. J Physiol. 1968 Mar;195(1):119–132. doi: 10.1113/jphysiol.1968.sp008450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Costantin L. L. The role of sodium current in the radial spread of contraction in frog muscle fibers. J Gen Physiol. 1970 Jun;55(6):703–715. doi: 10.1085/jgp.55.6.703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. DODGE F. A., FRANKENHAEUSER B. Membrane currents in isolated frog nerve fibre under voltage clamp conditions. J Physiol. 1958 Aug 29;143(1):76–90. doi: 10.1113/jphysiol.1958.sp006045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. DODGE F. A., FRANKENHAEUSER B. Sodium currents in the myelinated nerve fibre of Xenopus laevis investigated with the voltage clamp technique. J Physiol. 1959 Oct;148:188–200. doi: 10.1113/jphysiol.1959.sp006281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. FALK G., FATT P. LINEAR ELECTRICAL PROPERTIES OF STRIATED MUSCLE FIBRES OBSERVED WITH INTRACELLULAR ELECTRODES. Proc R Soc Lond B Biol Sci. 1964 Apr 14;160:69–123. doi: 10.1098/rspb.1964.0030. [DOI] [PubMed] [Google Scholar]
  16. FRANKENHAEUSER B. A method for recording resting and action potentials in the isolated myelinated nerve fibre of the frog. J Physiol. 1957 Mar 11;135(3):550–559. doi: 10.1113/jphysiol.1957.sp005729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Frankenhaeuser B., Lindley B. D., Smith R. S. Potentiometric measurement of membrane action potentials in frog muscle fibres. J Physiol. 1966 Mar;183(1):152–166. doi: 10.1113/jphysiol.1966.sp007857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. González-Serratos H. Inward spread of activation in vertebrate muscle fibres. J Physiol. 1971 Feb;212(3):777–799. doi: 10.1113/jphysiol.1971.sp009356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. HODGKIN A. L., HUXLEY A. F., KATZ B. Measurement of current-voltage relations in the membrane of the giant axon of Loligo. J Physiol. 1952 Apr;116(4):424–448. doi: 10.1113/jphysiol.1952.sp004716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hille B. The permeability of the sodium channel to organic cations in myelinated nerve. J Gen Physiol. 1971 Dec;58(6):599–619. doi: 10.1085/jgp.58.6.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hodgkin A. L., Nakajima S. Analysis of the membrane capacity in frog muscle. J Physiol. 1972 Feb;221(1):121–136. doi: 10.1113/jphysiol.1972.sp009743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hodgkin A. L., Nakajima S. The effect of diameter on the electrical constants of frog skeletal muscle fibres. J Physiol. 1972 Feb;221(1):105–120. doi: 10.1113/jphysiol.1972.sp009742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ildefonse M., Rougier O. Voltage-clamp analysis of the early current in frog skeletal muscle fibre using the double sucrose-gap method. J Physiol. 1972 Apr;222(2):373–395. doi: 10.1113/jphysiol.1972.sp009803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kao C. Y., Stanfield P. R. Actions of some anions on electrical properties and mechanical threshold of frog twitch muscle. J Physiol. 1968 Sep;198(2):291–309. doi: 10.1113/jphysiol.1968.sp008607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Keynes R. D., Rojas E. Kinetics and steady-state properties of the charged system controlling sodium conductance in the squid giant axon. J Physiol. 1974 Jun;239(2):393–434. doi: 10.1113/jphysiol.1974.sp010575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Moore J. W., Ramón F., Joyner R. W. Axon voltage-clamp simulations. I. Methods and tests. Biophys J. 1975 Jan;15(1):11–24. doi: 10.1016/S0006-3495(75)85788-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moore J. W., Ramón F., Joyner R. W. Axon voltage-clamp simulations. II. Double sucrose-gap method. Biophys J. 1975 Jan;15(1):25–35. doi: 10.1016/S0006-3495(75)85789-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Moore L. E. Voltage clamp experiments of single muscle fibers of Rana pipiens. J Gen Physiol. 1972 Jul;60(1):1–19. doi: 10.1085/jgp.60.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nakajima S., Bastian J. Double sucrose-gap method applied to single muscle fiber of Xenopus laevis. J Gen Physiol. 1974 Feb;63(2):235–256. doi: 10.1085/jgp.63.2.235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nonner W. A new voltage clamp method for Ranvier nodes. Pflugers Arch. 1969;309(2):176–192. doi: 10.1007/BF00586967. [DOI] [PubMed] [Google Scholar]
  31. Ramón F., Anderson N., Joyner R. W., Moore J. W. Axon voltage-clamp simulations. A multicellular preparation. Biophys J. 1975 Jan;15(1):55–69. doi: 10.1016/S0006-3495(75)85791-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schneider M. F. Linear electrical properties of the transverse tubules and surface membrane of skeletal muscle fibers. J Gen Physiol. 1970 Nov;56(5):640–671. doi: 10.1085/jgp.56.5.640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stanfield P. R. The onset of the effects of zinc and tetraethylammonium ions on action potential duration and twitch amplitude of single muscle fibres. J Physiol. 1973 Dec;235(3):639–654. doi: 10.1113/jphysiol.1973.sp010409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Valdiosera R., Clausen C., Eisenberg R. S. Impedance of frog skeletal muscle fibers in various solutions. J Gen Physiol. 1974 Apr;63(4):460–491. doi: 10.1085/jgp.63.4.460. [DOI] [PMC free article] [PubMed] [Google Scholar]

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