Skip to main content
PMC home page
The Journal of Physiology logoLink to The Journal of Physiology
. 1985 Dec;369:475–499. doi: 10.1113/jphysiol.1985.sp015911

Differential expression of inward and outward potassium currents in the macrophage-like cell line J774.1.

E K Gallin, P A Sheehy
PMCID: PMC1192659  PMID: 2419551

Abstract

J774.1 cells, a mouse-derived macrophage-like tumour cell line, were voltage clamped using whole-cell patch-clamp techniques. Cells were maintained in suspension cultures and plated at varying times before recording. The average zero-current potential of long-term adherent (greater than 24 h) cells was -77.6 mV. A tenfold increase in [K]o produced a 49 mV shift in zero-current potential. Freshly plated cells (less than 24 h) expressed two voltage-dependent currents: an outward current expressed transiently from 1 to 12 h post-plating and an inward current expressed 2-4 h post-plating which persisted in 100% of long-term adherent cells. Inward current was dependent upon voltage, time and [K]o 1/2, similar to the anomalous rectifier of other tissues. The conductance activated at potentials negative to -50 mV and plateaued at potentials negative to -110 mV. Inactivation was evident at potentials negative to -100 mV. Both the rate and extent of inactivation increased with hyperpolarization. Inward rectification was blocked by external BaCl2 or CsCl. The outward current was time- and voltage-dependent. The instantaneous I/V curves derived from tail experiments reversed at the potassium equilibrium potential (EK). A tenfold change of [K]o shifted the reversal potential 52 mV, indicating that the current was carried by potassium. This conductance activated at potentials positive to -50 mV, plateaued at potentials positive to -10 mV and inactivated completely with an exponential time course at all potentials. At voltages positive to -25 mV the rate of inactivation was independent of voltage. The outward current was blocked by 4-aminopyridine or D600. During the first 10 min after attaining a whole-cell recording, the conductance/voltage relation of the outward current shifted to more negative voltages and peak conductance showed a slight increase; recordings then stabilized. The voltage dependence of the inward current did not shift with time but wash-out of inward current was observed in some cells. The J774.1 cell line can serve as a model for the study of the role of voltage-dependent ionic conductances in macrophages.

Full text

PDF

Selected References

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

  1. Almers W. Potassium conductance changes in skeletal muscle and the potassium concentration in the transverse tubules. J Physiol. 1972 Aug;225(1):33–56. doi: 10.1113/jphysiol.1972.sp009928. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Almers W. The decline of potassium permeability during extreme hyperpolarization in frog skeletal muscle. J Physiol. 1972 Aug;225(1):57–83. doi: 10.1113/jphysiol.1972.sp009929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berton G., Gordon S. Superoxide release by peritoneal and bone marrow-derived mouse macrophages. Modulation by adherence and cell activation. Immunology. 1983 Aug;49(4):693–704. [PMC free article] [PubMed] [Google Scholar]
  4. Bloom B. R., Diamond B., Muschel R., Rosen N., Schneck J., Damiani G., Rosen O., Scharff M. Genetic approaches to the mechanism of macrophage functions. Fed Proc. 1978 Nov;37(13):2765–2771. [PubMed] [Google Scholar]
  5. Cahalan M. D., Chandy K. G., DeCoursey T. E., Gupta S. A voltage-gated potassium channel in human T lymphocytes. J Physiol. 1985 Jan;358:197–237. doi: 10.1113/jphysiol.1985.sp015548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chandy K. G., DeCoursey T. E., Cahalan M. D., McLaughlin C., Gupta S. Voltage-gated potassium channels are required for human T lymphocyte activation. J Exp Med. 1984 Aug 1;160(2):369–385. doi: 10.1084/jem.160.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cohen M. S., Ryan J. L., Root R. K. The oxidative metabolism of thioglycollate-elicited mouse peritoneal macrophages: the relationship between oxygen, superoxide and hydrogen peroxide and the effect of monolayer formation. J Immunol. 1981 Sep;127(3):1007–1011. [PubMed] [Google Scholar]
  8. Damiani G., Kiyotaki C., Soeller W., Sasada M., Peisach J., Bloom B. R. Macrophage variants in oxygen metabolism. J Exp Med. 1980 Oct 1;152(4):808–822. doi: 10.1084/jem.152.4.808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DeCoursey T. E., Chandy K. G., Gupta S., Cahalan M. D. Voltage-gated K+ channels in human T lymphocytes: a role in mitogenesis? Nature. 1984 Feb 2;307(5950):465–468. doi: 10.1038/307465a0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Fernandez J. M., Fox A. P., Krasne S. Membrane patches and whole-cell membranes: a comparison of electrical properties in rat clonal pituitary (GH3) cells. J Physiol. 1984 Nov;356:565–585. doi: 10.1113/jphysiol.1984.sp015483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fujimoto M., Kubota T. Physicochemical properties of a liquid ion exchanger microelectrode and its application to biological fluids. Jpn J Physiol. 1976;26(6):631–650. doi: 10.2170/jjphysiol.26.631. [DOI] [PubMed] [Google Scholar]
  13. Fukushima Y., Hagiwara S., Henkart M. Potassium current in clonal cytotoxic T lymphocytes from the mouse. J Physiol. 1984 Jun;351:645–656. doi: 10.1113/jphysiol.1984.sp015268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fukushima Y., Hagiwara S., Saxton R. E. Variation of calcium current during the cell growth cycle in mouse hybridoma lines secreting immunoglobulins. J Physiol. 1984 Oct;355:313–321. doi: 10.1113/jphysiol.1984.sp015421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gallin E. K. Calcium- and voltage-activated potassium channels in human macrophages. Biophys J. 1984 Dec;46(6):821–825. doi: 10.1016/S0006-3495(84)84080-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gallin E. K., Gallin J. I. Interaction of chemotactic factors with human macrophages. Induction of transmembrane potential changes. J Cell Biol. 1977 Oct;75(1):277–289. doi: 10.1083/jcb.75.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gallin E. K., Livengood D. R. Inward rectification in mouse macrophages: evidence for a negative resistance region. Am J Physiol. 1981 Jul;241(1):C9–17. doi: 10.1152/ajpcell.1981.241.1.C9. [DOI] [PubMed] [Google Scholar]
  18. Gallin E. K. Voltage clamp studies in macrophages from mouse spleen cultures. Science. 1981 Oct 23;214(4519):458–460. doi: 10.1126/science.7291986. [DOI] [PubMed] [Google Scholar]
  19. Gallin E. K., Wiederhold M. L., Lipsky P. E., Rosenthal A. S. Spontaneous and induced membrane hyperpolarizations in macrophages. J Cell Physiol. 1975 Dec;86 (Suppl 2)(3 Pt 2):653–661. doi: 10.1002/jcp.1040860510. [DOI] [PubMed] [Google Scholar]
  20. Hagiwara S., Miyazaki S., Rosenthal N. P. Potassium current and the effect of cesium on this current during anomalous rectification of the egg cell membrane of a starfish. J Gen Physiol. 1976 Jun;67(6):621–638. doi: 10.1085/jgp.67.6.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hagiwara S., Takahashi K. The anomalous rectification and cation selectivity of the membrane of a starfish egg cell. J Membr Biol. 1974;18(1):61–80. doi: 10.1007/BF01870103. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Lazdins J. K., Koech D. K., Karnovsky M. L. Oxidation of glucose by mouse peritoneal macrophages: a comparison of suspensions and monolayers. J Cell Physiol. 1980 Nov;105(2):191–196. doi: 10.1002/jcp.1041050202. [DOI] [PubMed] [Google Scholar]
  24. Leech C. A., Stanfield P. R. Inward rectification in frog skeletal muscle fibres and its dependence on membrane potential and external potassium. J Physiol. 1981;319:295–309. doi: 10.1113/jphysiol.1981.sp013909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Matteson D. R., Deutsch C. K channels in T lymphocytes: a patch clamp study using monoclonal antibody adhesion. Nature. 1984 Feb 2;307(5950):468–471. doi: 10.1038/307468a0. [DOI] [PubMed] [Google Scholar]
  26. Ohmori H. Inactivation kinetics and steady-state current noise in the anomalous rectifier of tunicate egg cell membranes. J Physiol. 1978 Aug;281:77–99. doi: 10.1113/jphysiol.1978.sp012410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Oliveira-Castro G. M., Dos Reis G. A. Electrophysiology of phagocytic membranes. III. Evidence for a calcium-dependent potassium permeability change during slow hyperpolarizations of activated macrophages. Biochim Biophys Acta. 1981 Jan 22;640(2):500–511. doi: 10.1016/0005-2736(81)90474-0. [DOI] [PubMed] [Google Scholar]
  28. Pofit J. F., Strauss P. R. Membrane transport by macrophages in suspension and adherent to glass. J Cell Physiol. 1977 Aug;92(2):249–255. doi: 10.1002/jcp.1040920213. [DOI] [PubMed] [Google Scholar]
  29. Ralph P., Nakoinz I. Phagocytosis and cytolysis by a macrophage tumour and its cloned cell line. Nature. 1975 Oct 2;257(5525):393–394. doi: 10.1038/257393a0. [DOI] [PubMed] [Google Scholar]
  30. Sakmann B., Trube G. Conductance properties of single inwardly rectifying potassium channels in ventricular cells from guinea-pig heart. J Physiol. 1984 Feb;347:641–657. doi: 10.1113/jphysiol.1984.sp015088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schwarze W., Kolb H. A. Voltage-dependent kinetics of an anionic channel of large unit conductance in macrophages and myotube membranes. Pflugers Arch. 1984 Nov;402(3):281–291. doi: 10.1007/BF00585511. [DOI] [PubMed] [Google Scholar]
  32. Snyderman R., Pike M. C., Fischer D. G., Koren H. S. Biologic and biochemical activities of continuous macrophage cell lines P388D1 and J774.1. J Immunol. 1977 Dec;119(6):2060–2066. [PubMed] [Google Scholar]
  33. Standen N. B., Stanfield P. R. Potassium depletion and sodium block of potassium currents under hyperpolarization in frog sartorius muscle. J Physiol. 1979 Sep;294:497–520. doi: 10.1113/jphysiol.1979.sp012943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Unkeless J. C., Kaplan G., Plutner H., Cohn Z. A. Fc-receptor variants of a mouse macrophage cell line. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1400–1404. doi: 10.1073/pnas.76.3.1400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ypey D. L., Clapham D. E. Development of a delayed outward-rectifying K+ conductance in cultured mouse peritoneal macrophages. Proc Natl Acad Sci U S A. 1984 May;81(10):3083–3087. doi: 10.1073/pnas.81.10.3083. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES