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. 1991 Aug;439:301–323. doi: 10.1113/jphysiol.1991.sp018668

Inactivating and non-inactivating dihydropyridine-sensitive Ca2+ channels in mouse cerebellar granule cells.

P A Slesinger 1, J B Lansman 1
PMCID: PMC1180110  PMID: 1654414

Abstract

1. Granule cells were dissociated from mouse cerebellum and grown in vitro. Currents through single Ca2+ channels were recorded from the cell body with the patch clamp technique. 2. Voltage steps to 0 mV produced brief channel openings with a mean open time of approximately 0.5 ms. The single-channel conductance measured from the amplitude of the single-channel current with 90 mM-Ba2+ in the patch electrode was 22 pS. 3. The probability of Ca2+ channel opening increased with test potentials more positive than -30 mV, with half-activation near 0 mV, and followed the Boltzmann relation for the activation of whole-cell Ca2+ current. 4. Voltage steps to potentials more positive than 0 mV produced more channel activity at the beginning than at the end of the voltage step. The average of the single-channel currents decayed to a non-zero level with a time course similar to that of the whole-cell Ca2+ current. 5. The amplitude as well as the decay of the mean current measured during a test pulse to 0 mV was reduced as the holding potential was made more positive than approximately -90 mV. The change in the open channel probability with holding potential followed the Boltzmann relation which described the inactivation of the whole-cell Ca2+ current. 6. Ca2+ channel activity persisted for over several minutes after excising the patch from the cell body when intracellular cyclic AMP was increased. After patch excision, the number of functional channels decreased to a level where only one channel at a time was active. Ca2+ channel openings appeared as either short bursts at the beginning of the voltage step or long bursts that lasted throughout the pulse. 7. Exposing the cell to the dihydropyridine agonist +(S)-202-791 markedly increased the fraction of sweeps with long openings and produced a non-decaying mean current that was approximately 5 times larger than control. In a fraction of the sweeps, however, long openings occurred more frequently at the beginning than at the end of the voltage step and these produced a decaying mean current. 8. Shifting the holding potential to more positive potentials in the presence of the dihydropyridine agonist preferentially reduced the number of brief openings while sparing the long openings. The amplitude of the mean current was similar to that obtained from the more negative holding potential and there was no change in the fraction of sweeps with long openings that occurred at the beginning of the voltage pulse.(ABSTRACT TRUNCATED AT 400 WORDS)

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Selected References

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

  1. Aldrich R. W., Corey D. P., Stevens C. F. A reinterpretation of mammalian sodium channel gating based on single channel recording. Nature. 1983 Dec 1;306(5942):436–441. doi: 10.1038/306436a0. [DOI] [PubMed] [Google Scholar]
  2. Aosaki T., Kasai H. Characterization of two kinds of high-voltage-activated Ca-channel currents in chick sensory neurons. Differential sensitivity to dihydropyridines and omega-conotoxin GVIA. Pflugers Arch. 1989 Jun;414(2):150–156. doi: 10.1007/BF00580957. [DOI] [PubMed] [Google Scholar]
  3. Armstrong D., Eckert R. Voltage-activated calcium channels that must be phosphorylated to respond to membrane depolarization. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2518–2522. doi: 10.1073/pnas.84.8.2518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bean B. P. Classes of calcium channels in vertebrate cells. Annu Rev Physiol. 1989;51:367–384. doi: 10.1146/annurev.ph.51.030189.002055. [DOI] [PubMed] [Google Scholar]
  5. Bean B. P. Nitrendipine block of cardiac calcium channels: high-affinity binding to the inactivated state. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6388–6392. doi: 10.1073/pnas.81.20.6388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brown A. M., Lux H. D., Wilson D. L. Activation and inactivation of single calcium channels in snail neurons. J Gen Physiol. 1984 May;83(5):751–769. doi: 10.1085/jgp.83.5.751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. COLE K. S., MOORE J. W. Potassium ion current in the squid giant axon: dynamic characteristic. Biophys J. 1960 Sep;1:1–14. doi: 10.1016/s0006-3495(60)86871-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Carbone E., Lux H. D. Single low-voltage-activated calcium channels in chick and rat sensory neurones. J Physiol. 1987 May;386:571–601. doi: 10.1113/jphysiol.1987.sp016552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chad J. E., Eckert R. An enzymatic mechanism for calcium current inactivation in dialysed Helix neurones. J Physiol. 1986 Sep;378:31–51. doi: 10.1113/jphysiol.1986.sp016206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cohen C. J., McCarthy R. T. Nimodipine block of calcium channels in rat anterior pituitary cells. J Physiol. 1987 Jun;387:195–225. doi: 10.1113/jphysiol.1987.sp016570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cull-Candy S. G., Howe J. R., Ogden D. C. Noise and single channels activated by excitatory amino acids in rat cerebellar granule neurones. J Physiol. 1988 Jun;400:189–222. doi: 10.1113/jphysiol.1988.sp017117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fenwick E. M., Marty A., Neher E. Sodium and calcium channels in bovine chromaffin cells. J Physiol. 1982 Oct;331:599–635. doi: 10.1113/jphysiol.1982.sp014394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fox A. P., Nowycky M. C., Tsien R. W. Kinetic and pharmacological properties distinguishing three types of calcium currents in chick sensory neurones. J Physiol. 1987 Dec;394:149–172. doi: 10.1113/jphysiol.1987.sp016864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fox A. P., Nowycky M. C., Tsien R. W. Single-channel recordings of three types of calcium channels in chick sensory neurones. J Physiol. 1987 Dec;394:173–200. doi: 10.1113/jphysiol.1987.sp016865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gurney A. M., Nerbonne J. M., Lester H. A. Photoinduced removal of nifedipine reveals mechanisms of calcium antagonist action on single heart cells. J Gen Physiol. 1985 Sep;86(3):353–379. doi: 10.1085/jgp.86.3.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  17. Hess P. Calcium channels in vertebrate cells. Annu Rev Neurosci. 1990;13:337–356. doi: 10.1146/annurev.ne.13.030190.002005. [DOI] [PubMed] [Google Scholar]
  18. Hess P., Lansman J. B., Tsien R. W. Calcium channel selectivity for divalent and monovalent cations. Voltage and concentration dependence of single channel current in ventricular heart cells. J Gen Physiol. 1986 Sep;88(3):293–319. doi: 10.1085/jgp.88.3.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hess P., Lansman J. B., Tsien R. W. Different modes of Ca channel gating behaviour favoured by dihydropyridine Ca agonists and antagonists. Nature. 1984 Oct 11;311(5986):538–544. doi: 10.1038/311538a0. [DOI] [PubMed] [Google Scholar]
  20. Hockberger P. E., Tseng H. Y., Connor J. A. Immunocytochemical and electrophysiological differentiation of rat cerebellar granule cells in explant cultures. J Neurosci. 1987 May;7(5):1370–1383. doi: 10.1523/JNEUROSCI.07-05-01370.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hof R. P., Rüegg U. T., Hof A., Vogel A. Stereoselectivity at the calcium channel: opposite action of the enantiomers of a 1,4-dihydropyridine. J Cardiovasc Pharmacol. 1985 Jul-Aug;7(4):689–693. doi: 10.1097/00005344-198507000-00012. [DOI] [PubMed] [Google Scholar]
  22. Hoshi T., Aldrich R. W. Gating kinetics of four classes of voltage-dependent K+ channels in pheochromocytoma cells. J Gen Physiol. 1988 Jan;91(1):107–131. doi: 10.1085/jgp.91.1.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hoshi T., Rothlein J., Smith S. J. Facilitation of Ca2+-channel currents in bovine adrenal chromaffin cells. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5871–5875. doi: 10.1073/pnas.81.18.5871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hoshi T., Smith S. J. Large depolarization induces long openings of voltage-dependent calcium channels in adrenal chromaffin cells. J Neurosci. 1987 Feb;7(2):571–580. doi: 10.1523/JNEUROSCI.07-02-00571.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kostyuk P. G., Shuba YaM, Savchenko A. N. Three types of calcium channels in the membrane of mouse sensory neurons. Pflugers Arch. 1988 Jun;411(6):661–669. doi: 10.1007/BF00580863. [DOI] [PubMed] [Google Scholar]
  26. Lacerda A. E., Brown A. M. Nonmodal gating of cardiac calcium channels as revealed by dihydropyridines. J Gen Physiol. 1989 Jun;93(6):1243–1273. doi: 10.1085/jgp.93.6.1243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lux H. D., Brown A. M. Patch and whole cell calcium currents recorded simultaneously in snail neurons. J Gen Physiol. 1984 May;83(5):727–750. doi: 10.1085/jgp.83.5.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Messer A. The maintenance and identification of mouse cerebellar granule cells in monolayer culture. Brain Res. 1977 Jul 8;130(1):1–12. doi: 10.1016/0006-8993(77)90838-1. [DOI] [PubMed] [Google Scholar]
  29. Miller R. J. Multiple calcium channels and neuronal function. Science. 1987 Jan 2;235(4784):46–52. doi: 10.1126/science.2432656. [DOI] [PubMed] [Google Scholar]
  30. Nowycky M. C., Fox A. P., Tsien R. W. Three types of neuronal calcium channel with different calcium agonist sensitivity. Nature. 1985 Aug 1;316(6027):440–443. doi: 10.1038/316440a0. [DOI] [PubMed] [Google Scholar]
  31. Ohmori H., Yoshii M. Surface potential reflected in both gating and permeation mechanisms of sodium and calcium channels of the tunicate egg cell membrane. J Physiol. 1977 May;267(2):429–463. doi: 10.1113/jphysiol.1977.sp011821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Plummer M. R., Logothetis D. E., Hess P. Elementary properties and pharmacological sensitivities of calcium channels in mammalian peripheral neurons. Neuron. 1989 May;2(5):1453–1463. doi: 10.1016/0896-6273(89)90191-8. [DOI] [PubMed] [Google Scholar]
  33. Sigworth F. J. The variance of sodium current fluctuations at the node of Ranvier. J Physiol. 1980 Oct;307:97–129. doi: 10.1113/jphysiol.1980.sp013426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Slesinger P. A., Lansman J. B. Inactivation of calcium currents in granule cells cultured from mouse cerebellum. J Physiol. 1991 Apr;435:101–121. doi: 10.1113/jphysiol.1991.sp018500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Somogyi P., Halasy K., Somogyi J., Storm-Mathisen J., Ottersen O. P. Quantification of immunogold labelling reveals enrichment of glutamate in mossy and parallel fibre terminals in cat cerebellum. Neuroscience. 1986 Dec;19(4):1045–1050. doi: 10.1016/0306-4522(86)90121-1. [DOI] [PubMed] [Google Scholar]
  36. Tsien R. W., Lipscombe D., Madison D. V., Bley K. R., Fox A. P. Multiple types of neuronal calcium channels and their selective modulation. Trends Neurosci. 1988 Oct;11(10):431–438. doi: 10.1016/0166-2236(88)90194-4. [DOI] [PubMed] [Google Scholar]
  37. White M. M., Bezanilla F. Activation of squid axon K+ channels. Ionic and gating current studies. J Gen Physiol. 1985 Apr;85(4):539–554. doi: 10.1085/jgp.85.4.539. [DOI] [PMC free article] [PubMed] [Google Scholar]

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