Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1991 Dec 15;88(24):11052–11056. doi: 10.1073/pnas.88.24.11052

Completely functional double-barreled chloride channel expressed from a single Torpedo cDNA.

C K Bauer 1, K Steinmeyer 1, J R Schwarz 1, T J Jentsch 1
PMCID: PMC53071  PMID: 1722317

Abstract

We have performed an electrophysiological analysis of the recently cloned Torpedo marmorata Cl- channel. Functional expression of Cl- channels in oocytes of Xenopus laevis previously injected with cRNA yielded an outward-rectifying current activated by hyperpolarization. Replacement of Cl- with other anions significantly reduced or inhibited the current. Single-channel recordings from cell-attached patches exhibited burst-like Cl- channel activity with rapid fluctuations between three equally spaced substates (0 pS, 9 pS, and 18 pS). The properties of the cloned Cl- channel were almost identical to those of the reconstituted native T. californica Cl- channel and were in full agreement with the predictions of the double-barreled channel model [Hanke, W. & Miller, C. (1983) J. Gen. Physiol. 82, 25-45]. Our results imply that the cloned cDNA codes for the completely functional Torpedo electroplax Cl- channel.

Full text

PDF
11052

Images in this article

11055Image
on p.11055

Selected References

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

  1. Barish M. E. A transient calcium-dependent chloride current in the immature Xenopus oocyte. J Physiol. 1983 Sep;342:309–325. doi: 10.1113/jphysiol.1983.sp014852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boton R., Dascal N., Gillo B., Lass Y. Two calcium-activated chloride conductances in Xenopus laevis oocytes permeabilized with the ionophore A23187. J Physiol. 1989 Jan;408:511–534. doi: 10.1113/jphysiol.1989.sp017473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chesnoy-Marchais D., Evans M. G. Chloride channels activated by hyperpolarization in Aplysia neurones. Pflugers Arch. 1986 Dec;407(6):694–696. doi: 10.1007/BF00582654. [DOI] [PubMed] [Google Scholar]
  4. Hanke W., Miller C. Single chloride channels from Torpedo electroplax. Activation by protons. J Gen Physiol. 1983 Jul;82(1):25–45. doi: 10.1085/jgp.82.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Jentsch T. J., Steinmeyer K., Schwarz G. Primary structure of Torpedo marmorata chloride channel isolated by expression cloning in Xenopus oocytes. Nature. 1990 Dec 6;348(6301):510–514. doi: 10.1038/348510a0. [DOI] [PubMed] [Google Scholar]
  6. Labarca P., Rice J. A., Fredkin D. R., Montal M. Kinetic analysis of channel gating. Application to the cholinergic receptor channel and the chloride channel from Torpedo californica. Biophys J. 1985 Apr;47(4):469–478. doi: 10.1016/S0006-3495(85)83939-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Matsuda H., Matsuura H., Noma A. Triple-barrel structure of inwardly rectifying K+ channels revealed by Cs+ and Rb+ block in guinea-pig heart cells. J Physiol. 1989 Jun;413:139–157. doi: 10.1113/jphysiol.1989.sp017646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Methfessel C., Witzemann V., Takahashi T., Mishina M., Numa S., Sakmann B. Patch clamp measurements on Xenopus laevis oocytes: currents through endogenous channels and implanted acetylcholine receptor and sodium channels. Pflugers Arch. 1986 Dec;407(6):577–588. doi: 10.1007/BF00582635. [DOI] [PubMed] [Google Scholar]
  9. Miledi R. A calcium-dependent transient outward current in Xenopus laevis oocytes. Proc R Soc Lond B Biol Sci. 1982 Jul 22;215(1201):491–497. doi: 10.1098/rspb.1982.0056. [DOI] [PubMed] [Google Scholar]
  10. Miller C. Open-state substructure of single chloride channels from Torpedo electroplax. Philos Trans R Soc Lond B Biol Sci. 1982 Dec 1;299(1097):401–411. doi: 10.1098/rstb.1982.0140. [DOI] [PubMed] [Google Scholar]
  11. Miller C., White M. M. A voltage-dependent chloride conductance channel from Torpedo electroplax membrane. Ann N Y Acad Sci. 1980;341:534–551. doi: 10.1111/j.1749-6632.1980.tb47197.x. [DOI] [PubMed] [Google Scholar]
  12. Miller C., White M. M. Dimeric structure of single chloride channels from Torpedo electroplax. Proc Natl Acad Sci U S A. 1984 May;81(9):2772–2775. doi: 10.1073/pnas.81.9.2772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sansom S. C., La B. Q., Carosi S. L. Double-barreled chloride channels of collecting duct basolateral membrane. Am J Physiol. 1990 Jul;259(1 Pt 2):F46–F52. doi: 10.1152/ajprenal.1990.259.1.F46. [DOI] [PubMed] [Google Scholar]
  14. Stocker M., Stühmer W., Wittka R., Wang X., Müller R., Ferrus A., Pongs O. Alternative Shaker transcripts express either rapidly inactivating or noninactivating K+ channels. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8903–8907. doi: 10.1073/pnas.87.22.8903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Taglietti V., Toselli M. A study of stretch-activated channels in the membrane of frog oocytes: interactions with Ca2+ ions. J Physiol. 1988 Dec;407:311–328. doi: 10.1113/jphysiol.1988.sp017417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. White M. M., Miller C. A voltage-gated anion channel from the electric organ of Torpedo californica. J Biol Chem. 1979 Oct 25;254(20):10161–10166. [PubMed] [Google Scholar]
  17. White M. M., Miller C. Probes of the conduction process of a voltage-gated Cl- channel from Torpedo electroplax. J Gen Physiol. 1981 Jul;78(1):1–18. doi: 10.1085/jgp.78.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES