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Elevation in intracellular calcium activates both chloride and proton currents in human macrophages

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Abstract

The transition of a resting macrophage into the activated state is accompanied by changes in membrane potential, cytoplasmic pH, and intracellular calcium (Ca i ). Activation of Cl− as well as H+-selective currents may give rise to stimulus-induced changes in membrane potential and counteract changes in intra-cellular pH (pH i ) which have been observed to be closely associated with respiratory burst activation and superoxide production in macrophages. We carried out whole-cell voltage clamp experiments on human monocyte-derived macrophages (HMDMs) and characterized currents activated following an elevation in Ca i using isosmotic pipette and bath solutions in which Cl− was the major permeant species. Ca i was elevated by exposing cells to the Ca2+ ionophore A23187 (1–10 μm) in the presence of extracellular Ca2+ or by internally exchanging the patch-electrode solution with ones buffered to free Ca2+ concentrations between 40 and 2,000 nm. We have identified two Ca2+-dependent ion conductances based on differences in their characteristic time-dependent kinetics: a rapidly activating Cl− conductance that showed variable inactivation at depolarized potentials and a H+ conductance with delayed activation kinetics. Both conductances were inhibited by the disulfonic acid stilbene DIDS (100 μm). Current activation for both Ca2+-dependent conductances was phosphorylation dependent, neither conductance appeared in the presence of the broad spectrum kinase inhibitor H-7 (75 μm). Inclusion of the autophosphorylated, Ca2+/calmodulin-dependent protein kinase in the pipette in the presence of ATP induced a rapidly activating current similar to that observed following an elevation in Ca i . Activation of both conductances would contribute to the changes in membrane potential which accompany stimulation-induced activation of macrophages as well as counteract the decrease in pH i during sustained Superoxide production.

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References

  • Amador, M., Dani, J. 1991. Protein kinase inhibitor, H-7, directly affects NMDA receptor channels. Neurosci. Lett. 124:251–255

    Google Scholar 

  • Barish, M.E., Baud, C. 1984. A voltage-gated hydrogen ion current in the oocyte membrane of the axolotl, Ambystoma. J. Physiol. 352:243–263

    Google Scholar 

  • Byerly, L., Meech, R., Moody, W. 1984. Rapidly activating hydrogen ion currents in perfused neurons of the snail, Lymnaea stagnalis. J. Physiol. 351:199–216

    Google Scholar 

  • Byerly, L., Suen, Y. 1989. Characterization of protein currents in neurons of the snail, Lymnaea stagnalis. J Physiol. 413:75–89

    Google Scholar 

  • Chan, H.-C., Goldstein, J., Nelson, D.J. 1992. Alternate pathways for chloride conductance activation in normal and cystic fibrosis air-way epithelial cells. Am. J. Physiol. 262:C1273-C1283

    Google Scholar 

  • Christensen, O. 1987. Mediation of cell volume regulation by Ca2+ influx through stretch-activated channels. Nature 330:66–68

    Google Scholar 

  • Conrad, G.W., Rink, T.J. 1986. Platelet activating factor raises intracellular calcium ion concentration in macrophages. J. Cell Biol. 103:439–450

    Google Scholar 

  • Decoursey, T.E. 1991. Hydrogen ion currents in rat alveolar epithelial cells. Biophys. J. 60:1243–1253

    Google Scholar 

  • Decoursey, T.E., Cherny, V. 1993. Potential, pH, and arachidonate gate hydrogen ion currents in human neutrophils. Biophys. J. 65:1590–1598

    Google Scholar 

  • Fabiato, A., Fabiato, F. 1979. Calculator programs for computing the composition of the solutions containing multiple metals and ligands used for experiments in skinned muscle cells. J. Physiol. (Paris) 79:463–505

    Google Scholar 

  • Gallin, E.K. 1984. Calcium- and voltage-activated potassium channels in human macrophages. Biophys. J. 46:821–825

    Google Scholar 

  • Gallin, E.K. 1989. Evidence for a Ca-activated inwardly rectifying K channel in human macrophages. Am. J. Physiol. 257:C77-C85

    Google Scholar 

  • Gallin, E.K., McKinney, L.C. 1988. Patch-clamp studies in human macrophages: Single channel and whole-cell characterization of two K+ conductances. J. Membrane Biol. 103:55–66

    Google Scholar 

  • Gray, L.S., Russell, J.H. 1986. Cytolytic T lymphocyte effector function requires plasma membrane chloride flux. J. Immunol. 136:3032–3037.

    Google Scholar 

  • Grissmer, S., Lewis, R.S., Cahalan, M.D. 1992. Ca2+-activated K+ channels in human leukemic T cells. J. Gen. Physiol. 99:63–84

    Google Scholar 

  • Hagiwara, S., Yoshii, M. 1979. Effects of internal potassium and sodium on the anomalous rectification of the starfish egg as examined by internal perfusion. J. Physiol. 292:251–265

    Google Scholar 

  • Hamill, O.P., Marty, A., Neher, E., Sakmann, B., Sigworth, F.J. 1981. Improved patch-clamp techniques for high resolution current recording from cells and cell-free membrane patches. Pfluegers Arch. 391:85–100

    Google Scholar 

  • Hara, N., Ichinose, M., Sawada, M., Moeno, T. 1991. The activation of Ca2+-dependent K+ conductance by adrenaline in mouse peritoneal macrophages. Pfluegers Arch. 419:371–379

    Google Scholar 

  • Hartley, Z., Dubinsky, J.M. 1993. Changes in intracellular pH associated with glutamate excitotoxicity. J. Neurosci. 13:4690–4699

    Google Scholar 

  • Henderson, L.M., Chappell, J.B., Jones, O.T.G. 1987. The superoxide-generating NADPH oxidase of human neutrophils is electrogenic and associated with an H+ channel. Biochem. J. 246:325–329

    Google Scholar 

  • Henderson, L.M., Chappell, J.B., Jones, O.T.G. 1988. Internal pH changes associated with the activity of NADPH oxidase of human neutrophils: further evidence for the presence of an H+ conducting channel. Biochem. J. 251:563–567

    Google Scholar 

  • Jaconi, M.E.E., Lew, D.P., Carpentier, J.-L., Magnusson, K.E., Sjögren, M., Stendahl, O. 1990. Cytosolic free calcium elevation mediates the phagosome-lysosome fusion during phagocytosis in human neutrophils. J. Cell Biol. 110:1555–1564

    Google Scholar 

  • Kapus, A., Szazi, K., Ligeti, E. 1992. Phorbol 12-myristate 13-acetate activates an electrogenic H+-conducting pathway in the membrane of neutrophils. Biochem. J. 281:697–701

    Google Scholar 

  • Katnik, C., Nelson, D.J. 1993. Platelet activating factor-induced increase in cytosolic calcium and transmembrane current in human macrophages. J. Membrane Biol. 134:213–224

    Google Scholar 

  • Kolb, H.-A., Ubl, J. 1987. Activation of anion channels by zymosan particles in membranes of peritoneal macrophages. Biochim. Biophys. Acta 899:239–246

    Google Scholar 

  • Krause, K.H., Welsh, M.J. 1990. Voltage-dependent and Ca2+-activated ion channels in human neutrophils. J. Clin. Invest. 85:491–498

    Google Scholar 

  • Kruskal, B.A., Maxfield, F.R. 1987. Cytosolic free calcium increases before and oscillates during frustrated phagocytosis in macrophages. J. Cell Biol. 105:2685–2693

    Google Scholar 

  • Lew, D.P., Andersson, T., Hed, J., Di Virgilio, F., Pozzan, T., Stendahl, O. 1985. Ca i -dependent and Ca2+-independent phagocytosis in human neutrophils. Nature 315:509–512

    Google Scholar 

  • Lipton, S.A. 1986. Antibody activates cationic channels via second messenger Ca2+. Biochim. Biophys. Acta 856:59–67

    Google Scholar 

  • Lukacs, G.L., Kapus, A., Nanda, A., Romanek, R., Grinstein, S. 1993. Proton conductance of the plasma membrane: properties, regulation, and functional role. Am. J. Physiol. 265:C3-C14

    Google Scholar 

  • Mahaut-Smith, M.P. 1990. Chloride channels in human platelets: evidence for activation by internal calcium. J. Membrane Biol. 118:69–75

    Google Scholar 

  • Mahaut-Smith, M.P., Mason M.J. 1991. Single channel recordings of Ca2+-activated K+ channels in rat thymic lymphocytes: activation by concanavalin A. J. Physiol. 439:513–528

    Google Scholar 

  • Matthews, G., Neher, E., Penner, R. 1989. Chloride conductance activated by external agonists and internal messengers in rat peritoneal mast cells. J. Physiol. 418:131–144

    Google Scholar 

  • McCarty, N.A., O'Neil, R.G. 1991a. Calcium-dependent control of volume regulation in renal proximal tubule cells: I. Swelling-activated Ca2+ entry and release. J. Membrane Biol. 123:149–160

    Google Scholar 

  • McCarty, N.A., O'Neil, R.G. 1991b. Calcium-dependent control ofvolume regulation in renal proximal tubule cells: II. Roles of dihydropyridine-sensitive and -insensitive Ca2+ entry pathways. J. Membrane Biol. 123:161–170

    Google Scholar 

  • Meech, R.W., Thomas, R.C. 1977. The effect of calcium injection on the intracellular sodium and pH of snail neurons. J. Physiol. 265:867–879

    Google Scholar 

  • Meech, R.W., Thomas, R.C. 1980. Effect of measured calcium chloride injections on the membrane potential and pH of snail neurons. J. Physiol. 298:111–129

    Google Scholar 

  • Montrose-Rafizadeh, C., Guggino, W. B. 1991. Role of intracellular calcium in volume regulation by rabbit medullary thick ascending limb cells. Am. J. Physiol. 260:F402-F409

    Google Scholar 

  • Nanda, A., Grinstein, S. 1991. Protein kinase C activates on H+ (equivalent) conductance in the plasma membrane of human neutrophils. Proc. Natl. Acad. Sci. USA 88:10816–10820

    Google Scholar 

  • Neher, E. 1988. The influence of intracellular calcium concentration on degranulation of dialysed mast cells from rat peritoneum. J. Physiol. 395:193–214

    Google Scholar 

  • Nelson, D.J., Jow, B., Jow, F. 1990. Whole-cell currents in macrophages: I. Human monocyte-derived macrophages. J. Membrane Biol. 117:29–44

    Google Scholar 

  • Nishimoto, I., Wagner, J.A., Schulman, H., Gardner, P. 1991. Regulation of Cl− channels by multifunctional CaM kinase. Neuron 6:547–555

    Google Scholar 

  • Randriamampita, C., Bismuth, G., Trautmann, A. 1991. Ca2+-induced Ca2+ release amplifies the Ca2+ response elicited by inositol trisphosphate in macrophages. Cell. Regul. 2:513–522

    Google Scholar 

  • Randriamampita, C., Trautmann, A. 1987. Ionic channels in murine macrophages. J. Cell Biol. 105:761–769

    Google Scholar 

  • Reuhl, T.O.K., Amador, A., Moorman, J.R., Pinkham, J., Dani, J.A. 1992. Nicotinic acetylcholine receptors are directly affected by agents used to study protein phosphorylation. J. Neurophysiol. 68:407–416

    Google Scholar 

  • Schumann, M.A., Gardner, P., Raffin, R.A. 1993. Recombinant human tumor necrosis factor α induces calcium oscillation and calcium-activated chloride current in human neutrophils. J. Biol. Chem. 268:2134–2140

    Google Scholar 

  • Soejima, M., Noma, A. 1984. Mode of regulation of the ACh-sensitive K-channel by the muscarinic receptor in rabbit atrial cells. Pfluegers Arch. 400:424–431

    Google Scholar 

  • Thomas, R.C. 1989. Proton channels in snail neurons. Does calcium entry mimic the effects of proton influx? Ann. NY Acad. Sci. 574:287–293

    Google Scholar 

  • Wagner, J.A., Cozens, A.L., Schulman, H., Gruenert, D.C., Stryer, L., Gardner, P. 1991. Activation of chloride channels in normal and cystic fibrosis airway epithelial cells by multifunctional calcium/calmodulin-dependent protein kinase. Nature 349:793–796

    Google Scholar 

  • Wong, S.M.E., Chase, H.S., Jr. 1986. Role of intracellular calcium in cellular volume regulation. Am. J. Physiol. 250:C841-C852

    Google Scholar 

  • Worrell, R.T., Frizzell, R.A. 1991. CaMKII mediates stimulation of chloride conductance by calcium in T84 cells. Am. J. Physiol. 260:C877-C882

    Google Scholar 

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Authors and Affiliations

  1. Departments of Medicine and Neurology and the Committee on Cell Physiology, The University of Chicago, 60637, Chicago, Illinois

    K. O. Holevinsky, F. Jow & D. J. Nelson

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  1. K. O. Holevinsky
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  2. F. Jow
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  3. D. J. Nelson
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Additional information

The authors wish to thank Dr. H. Schulman for providing us with the purified CaMKII and Jennifer Foss for technical assistance. This work was supported by National Institutes of Health RO1 GM36823.

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Holevinsky, K.O., Jow, F. & Nelson, D.J. Elevation in intracellular calcium activates both chloride and proton currents in human macrophages. J. Membarin Biol. 140, 13–30 (1994). https://doi.org/10.1007/BF00234482

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  • DOI: https://doi.org/10.1007/BF00234482

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