Abstract
1. A voltage-dependent K+ channel called the 'n' type (for 'normal') is the most prevalent ion channel found in whole-cell recordings from T lymphocytes. In whole-cell patch-clamp recordings activity of the n-type channel is affected by mitogenic agents, pH, Ca2+ and temperature but not by cyclic nucleotides. Because channel properties and regulation can depend on cytoplasmic components we sought to reassess the properties of K+ channels in intact, normal human T lymphocytes using cell-attached, patch-clamp recordings. In the present study, we show that the predominant K+ channel in resting, intact cells is the n type and is affected by voltage, temperature and Ca2+ in ways similar to the disrupted cell. Moreover, K+ channels are activated by agents that raise cyclic AMP in intact cells. 2. In cell-attached recordings, we found voltage-activated K+ channels in about 60% of patches at room temperature. The channel was K+ selective as judged from the reversal potential under different Ka(+)-K+ gradients and at different resting membrane potentials. Some patches were subsequently excised and the selectivity further confirmed. The current-voltage relation was inwardly rectifying under symmetrical K+ concentrations and had a slope conductance of 9.4 pS at 50 mV depolarized and 23.8 pS at 50 mV hyperpolarized from the resting potential. From the reversal potentials under various conditions the cell resting potential was -51 +/- 1 mV in normal NaCl saline and about 0 mV when the bath contained 150 mM-KCl saline. Two other types of K+ channel were seen in resting, intact cells, but were much less common (less than 5% and 11% of patches). A large-conductance K+ channel was seen in less than 1% of inside-out patches. 3. The predominant K+ channel in intact, resting T lymphocytes was confirmed as the n type underlying the whole-cell K+ current evoked by voltage steps. In cell-attached patches there was a low, steady-state level of activity at the resting potential but activity was greatly increased by depolarizing voltage jumps. Steady-state inactivation could be removed by a hyperpolarizing pre-pulse. Ensemble currents constructed by summing channel openings during repeated voltage jumps showed sigmoid kinetics of current activation and a monoexponential decay phase. These kinetics were well fitted by a Hodgkin-Huxley-type n4j kinetic model with time constants very similar to the whole-cell current of disrupted cells. Moreover, the kinetics depended on the external K+ concentration as previous research has shown.(ABSTRACT TRUNCATED AT 400 WORDS)
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Selected References
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- Bregestovski P., Redkozubov A., Alexeev A. Elevation of intracellular calcium reduces voltage-dependent potassium conductance in human T cells. 1986 Feb 27-Mar 5Nature. 319(6056):776–778. doi: 10.1038/319776a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Cavagnaro J., Lewis R. M. Bidirectional regulatory circuit between the immune and neuroendocrine systems. Year Immunol. 1989;4:241–252. [PubMed] [Google Scholar]
- Chen J. H., Schulman H., Gardner P. A cAMP-regulated chloride channel in lymphocytes that is affected in cystic fibrosis. Science. 1989 Feb 3;243(4891):657–660. doi: 10.1126/science.2464852. [DOI] [PubMed] [Google Scholar]
- Choquet D., Korn H. Modulation of voltage-dependent potassium channels in B lymphocytes. Biochem Pharmacol. 1988 Oct 15;37(20):3797–3802. doi: 10.1016/0006-2952(88)90058-5. [DOI] [PubMed] [Google Scholar]
- Choquet D., Sarthou P., Primi D., Cazenave P. A., Korn H. Cyclic AMP-modulated potassium channels in murine B cells and their precursors. Science. 1987 Mar 6;235(4793):1211–1214. doi: 10.1126/science.2434998. [DOI] [PubMed] [Google Scholar]
- Dailey M. O., Schreurs J., Schulman H. Hormone receptors on cloned T lymphocytes. Increased responsiveness to histamine, prostaglandins, and beta-adrenergic agents as a late stage event in T cell activation. J Immunol. 1988 May 1;140(9):2931–2936. [PubMed] [Google Scholar]
- 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]
- DeCoursey T. E. State-dependent inactivation of K+ currents in rat type II alveolar epithelial cells. J Gen Physiol. 1990 Apr;95(4):617–646. doi: 10.1085/jgp.95.4.617. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gardner P. Patch clamp studies of lymphocyte activation. Annu Rev Immunol. 1990;8:231–252. doi: 10.1146/annurev.iy.08.040190.001311. [DOI] [PubMed] [Google Scholar]
- Grinstein S., Dixon S. J. Ion transport, membrane potential, and cytoplasmic pH in lymphocytes: changes during activation. Physiol Rev. 1989 Apr;69(2):417–481. doi: 10.1152/physrev.1989.69.2.417. [DOI] [PubMed] [Google Scholar]
- Grinstein S., Rothstein A., Sarkadi B., Gelfand E. W. Responses of lymphocytes to anisotonic media: volume-regulating behavior. Am J Physiol. 1984 Mar;246(3 Pt 1):C204–C215. doi: 10.1152/ajpcell.1984.246.3.C204. [DOI] [PubMed] [Google Scholar]
- Kao J. P., Harootunian A. T., Tsien R. Y. Photochemically generated cytosolic calcium pulses and their detection by fluo-3. J Biol Chem. 1989 May 15;264(14):8179–8184. [PubMed] [Google Scholar]
- Krause D., Lee S. C., Deutsch C. Forskolin effects on the voltage-gated K+ conductance of human T cells. Pflugers Arch. 1988 Jul;412(1-2):133–140. doi: 10.1007/BF00583742. [DOI] [PubMed] [Google Scholar]
- Lee S. C., Deutsch C. Temperature dependence of K(+)-channel properties in human T lymphocytes. Biophys J. 1990 Jan;57(1):49–62. doi: 10.1016/S0006-3495(90)82506-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levitan I. B. Modulation of ion channels in neurons and other cells. Annu Rev Neurosci. 1988;11:119–136. doi: 10.1146/annurev.ne.11.030188.001003. [DOI] [PubMed] [Google Scholar]
- Lewis R. S., Cahalan M. D. Subset-specific expression of potassium channels in developing murine T lymphocytes. Science. 1988 Feb 12;239(4841 Pt 1):771–775. doi: 10.1126/science.2448877. [DOI] [PubMed] [Google Scholar]
- Mahaut-Smith M. P., Schlichter L. C. Ca2(+)-activated K+ channels in human B lymphocytes and rat thymocytes. J Physiol. 1989 Aug;415:69–83. doi: 10.1113/jphysiol.1989.sp017712. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Pahapill P. A., Schlichter L. C. Modulation of potassium channels in human T lymphocytes: effects of temperature. J Physiol. 1990 Mar;422:103–126. doi: 10.1113/jphysiol.1990.sp017975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Plaut M. Lymphocyte hormone receptors. Annu Rev Immunol. 1987;5:621–669. doi: 10.1146/annurev.iy.05.040187.003201. [DOI] [PubMed] [Google Scholar]
- Schlichter L. C., Grygorczyk R., Pahapill P. A., Grygorczyk C. A large, multiple-conductance chloride channel in normal human T lymphocytes. Pflugers Arch. 1990 Jun;416(4):413–421. doi: 10.1007/BF00370748. [DOI] [PubMed] [Google Scholar]
- Schlichter L., Sidell N., Hagiwara S. K channels are expressed early in human T-cell development. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5625–5629. doi: 10.1073/pnas.83.15.5625. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schlichter L., Sidell N., Hagiwara S. Potassium channels mediate killing by human natural killer cells. Proc Natl Acad Sci U S A. 1986 Jan;83(2):451–455. doi: 10.1073/pnas.83.2.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsien R. Y., Pozzan T., Rink T. J. T-cell mitogens cause early changes in cytoplasmic free Ca2+ and membrane potential in lymphocytes. Nature. 1982 Jan 7;295(5844):68–71. doi: 10.1038/295068a0. [DOI] [PubMed] [Google Scholar]
