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. 1998 Oct;75(4):1669–1678. doi: 10.1016/S0006-3495(98)77609-X

Calcium waves induced by large voltage pulses in fish keratocytes.

I Brust-Mascher 1, W W Webb 1
PMCID: PMC1299839  PMID: 9746509

Abstract

Intracellular calcium waves in fish keratocytes are induced by the application of electric field pulses with amplitudes between 55 and 120 V/cm and full width at half-maximum of 65-100 ms. Calcium concentrations were imaged using two-photon excited fluorescence microscopy (Denk et al., 1990 Science. 248:73-76; Williams et al. 1994 FASEB J. 8:804-813) and the ratiometric calcium indicator indo-1. The applied electric field pulses induced waves with fast calcium rise times and slow decays, which nucleated in the lamellipodium at the hyperpolarized side of the cells and, less frequently, at the depolarized side. The effectiveness of wave generation was determined by the change induced in the membrane potential, which is about half the field strength times the cell width in the direction of the field. Stimulation of waves began at voltage drops across the cell above 150 mV and saturated at voltage drops above 300 mV, where almost all cells exhibited a wave. Waves were not induced in low-calcium media and were blocked by the nonselective calcium channel blockers cobalt chloride and verapamil, but not by specific organic antagonists of voltage-sensitive calcium channel conductance. Thapsigargin stopped wave propagation in the cell body, indicating that calcium release from intracellular stores is necessary. Thus a voltage pulse stimulates Ca2+ influx through calcium channels in the plasma membrane, and if the intracellular calcium concentration reaches a threshold, release from intracellular stores is induced, creating a propagating wave. These observations and the measured parameters (average velocity approximately 66 micron/s and average rise time approximately 68 ms) are consistent with a wave amplification model in which[equation, see text] determines the effective diffusivity of the propagating molecules, D approximately 300 micron2/s (Meyer, 1991. Cell. 64:675-678).

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Selected References

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  1. Allbritton N. L., Meyer T., Stryer L. Range of messenger action of calcium ion and inositol 1,4,5-trisphosphate. Science. 1992 Dec 11;258(5089):1812–1815. doi: 10.1126/science.1465619. [DOI] [PubMed] [Google Scholar]
  2. Atri A., Amundson J., Clapham D., Sneyd J. A single-pool model for intracellular calcium oscillations and waves in the Xenopus laevis oocyte. Biophys J. 1993 Oct;65(4):1727–1739. doi: 10.1016/S0006-3495(93)81191-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bassani J. W., Bassani R. A., Bers D. M. Calibration of indo-1 and resting intracellular [Ca]i in intact rabbit cardiac myocytes. Biophys J. 1995 Apr;68(4):1453–1460. doi: 10.1016/S0006-3495(95)80318-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berridge M. J. Inositol trisphosphate and calcium signalling. Nature. 1993 Jan 28;361(6410):315–325. doi: 10.1038/361315a0. [DOI] [PubMed] [Google Scholar]
  5. Blatter L. A., Wier W. G. Intracellular diffusion, binding, and compartmentalization of the fluorescent calcium indicators indo-1 and fura-2. Biophys J. 1990 Dec;58(6):1491–1499. doi: 10.1016/S0006-3495(90)82494-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brown M. J., Loew L. M. Electric field-directed fibroblast locomotion involves cell surface molecular reorganization and is calcium independent. J Cell Biol. 1994 Oct;127(1):117–128. doi: 10.1083/jcb.127.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brundage R. A., Fogarty K. E., Tuft R. A., Fay F. S. Calcium gradients underlying polarization and chemotaxis of eosinophils. Science. 1991 Nov 1;254(5032):703–706. doi: 10.1126/science.1948048. [DOI] [PubMed] [Google Scholar]
  8. Brundage R. A., Fogarty K. E., Tuft R. A., Fay F. S. Chemotaxis of newt eosinophils: calcium regulation of chemotactic response. Am J Physiol. 1993 Dec;265(6 Pt 1):C1527–C1543. doi: 10.1152/ajpcell.1993.265.6.C1527. [DOI] [PubMed] [Google Scholar]
  9. Cheer A., Vincent J. P., Nuccitelli R., Oster G. Cortical activity in vertebrate eggs. I: The activation waves. J Theor Biol. 1987 Feb 21;124(4):377–404. doi: 10.1016/s0022-5193(87)80217-5. [DOI] [PubMed] [Google Scholar]
  10. Clapham D. E. Calcium signaling. Cell. 1995 Jan 27;80(2):259–268. doi: 10.1016/0092-8674(95)90408-5. [DOI] [PubMed] [Google Scholar]
  11. Cooper M. S., Schliwa M. Electrical and ionic controls of tissue cell locomotion in DC electric fields. J Neurosci Res. 1985;13(1-2):223–244. doi: 10.1002/jnr.490130116. [DOI] [PubMed] [Google Scholar]
  12. Cooper M. S., Schliwa M. Motility of cultured fish epidermal cells in the presence and absence of direct current electric fields. J Cell Biol. 1986 Apr;102(4):1384–1399. doi: 10.1083/jcb.102.4.1384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. De Young G. W., Keizer J. A single-pool inositol 1,4,5-trisphosphate-receptor-based model for agonist-stimulated oscillations in Ca2+ concentration. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9895–9899. doi: 10.1073/pnas.89.20.9895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Denk W., Strickler J. H., Webb W. W. Two-photon laser scanning fluorescence microscopy. Science. 1990 Apr 6;248(4951):73–76. doi: 10.1126/science.2321027. [DOI] [PubMed] [Google Scholar]
  15. Feder T. J., Webb W. W. Redistribution of plasma membrane proteins by electroosmosis elicits cytosolic calcium response in tumor mast cells. J Cell Physiol. 1994 Nov;161(2):227–236. doi: 10.1002/jcp.1041610206. [DOI] [PubMed] [Google Scholar]
  16. Fewtrell C. Ca2+ oscillations in non-excitable cells. Annu Rev Physiol. 1993;55:427–454. doi: 10.1146/annurev.ph.55.030193.002235. [DOI] [PubMed] [Google Scholar]
  17. Forscher P. Calcium and polyphosphoinositide control of cytoskeletal dynamics. Trends Neurosci. 1989 Nov;12(11):468–474. doi: 10.1016/0166-2236(89)90098-2. [DOI] [PubMed] [Google Scholar]
  18. Ghosh P. M., Keese C. R., Giaever I. Monitoring electropermeabilization in the plasma membrane of adherent mammalian cells. Biophys J. 1993 May;64(5):1602–1609. doi: 10.1016/S0006-3495(93)81531-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Girard S., Clapham D. Acceleration of intracellular calcium waves in Xenopus oocytes by calcium influx. Science. 1993 Apr 9;260(5105):229–232. doi: 10.1126/science.8385801. [DOI] [PubMed] [Google Scholar]
  20. Gross D., Loew L. M., Webb W. W. Optical imaging of cell membrane potential changes induced by applied electric fields. Biophys J. 1986 Aug;50(2):339–348. doi: 10.1016/S0006-3495(86)83467-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  22. Hahn K., DeBiasio R., Taylor D. L. Patterns of elevated free calcium and calmodulin activation in living cells. Nature. 1992 Oct 22;359(6397):736–738. doi: 10.1038/359736a0. [DOI] [PubMed] [Google Scholar]
  23. Hallett M., Carbone E. Studies of calcium influx into squid giant axons with aequorin. J Cell Physiol. 1972 Oct;80(2):219–226. doi: 10.1002/jcp.1040800208. [DOI] [PubMed] [Google Scholar]
  24. Hendey B., Maxfield F. R. Regulation of neutrophil motility and adhesion by intracellular calcium transients. Blood Cells. 1993;19(1):143–164. [PubMed] [Google Scholar]
  25. Hove-Madsen L., Bers D. M. Indo-1 binding to protein in permeabilized ventricular myocytes alters its spectral and Ca binding properties. Biophys J. 1992 Jul;63(1):89–97. doi: 10.1016/S0006-3495(92)81597-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Jaffe L. F. The path of calcium in cytosolic calcium oscillations: a unifying hypothesis. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9883–9887. doi: 10.1073/pnas.88.21.9883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Janmey P. A. Phosphoinositides and calcium as regulators of cellular actin assembly and disassembly. Annu Rev Physiol. 1994;56:169–191. doi: 10.1146/annurev.ph.56.030194.001125. [DOI] [PubMed] [Google Scholar]
  28. Lückhoff A., Clapham D. E. Calcium channels activated by depletion of internal calcium stores in A431 cells. Biophys J. 1994 Jul;67(1):177–182. doi: 10.1016/S0006-3495(94)80467-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Marks A. R. Intracellular calcium-release channels: regulators of cell life and death. Am J Physiol. 1997 Feb;272(2 Pt 2):H597–H605. doi: 10.1152/ajpheart.1997.272.2.H597. [DOI] [PubMed] [Google Scholar]
  30. Marks P. W., Hendey B., Maxfield F. R. Attachment to fibronectin or vitronectin makes human neutrophil migration sensitive to alterations in cytosolic free calcium concentration. J Cell Biol. 1991 Jan;112(1):149–158. doi: 10.1083/jcb.112.1.149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Marks P. W., Maxfield F. R. Transient increases in cytosolic free calcium appear to be required for the migration of adherent human neutrophils. J Cell Biol. 1990 Jan;110(1):43–52. doi: 10.1083/jcb.110.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Maxfield F. R. Regulation of leukocyte locomotion by Ca2+. Trends Cell Biol. 1993 Nov;3(11):386–391. doi: 10.1016/0962-8924(93)90088-i. [DOI] [PubMed] [Google Scholar]
  33. McCloskey M. A., Liu Z. Y., Poo M. M. Lateral electromigration and diffusion of Fc epsilon receptors on rat basophilic leukemia cells: effects of IgE binding. J Cell Biol. 1984 Sep;99(3):778–787. doi: 10.1083/jcb.99.3.778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Meyer T. Cell signaling by second messenger waves. Cell. 1991 Feb 22;64(4):675–678. doi: 10.1016/0092-8674(91)90496-l. [DOI] [PubMed] [Google Scholar]
  35. Meyer T., Stryer L. Calcium spiking. Annu Rev Biophys Biophys Chem. 1991;20:153–174. doi: 10.1146/annurev.bb.20.060191.001101. [DOI] [PubMed] [Google Scholar]
  36. Mittal A. K., Bereiter-Hahn J. Ionic control of locomotion and shape of epithelial cells: I. Role of calcium influx. Cell Motil. 1985;5(2):123–136. doi: 10.1002/cm.970050205. [DOI] [PubMed] [Google Scholar]
  37. Miyazaki S., Yuzaki M., Nakada K., Shirakawa H., Nakanishi S., Nakade S., Mikoshiba K. Block of Ca2+ wave and Ca2+ oscillation by antibody to the inositol 1,4,5-trisphosphate receptor in fertilized hamster eggs. Science. 1992 Jul 10;257(5067):251–255. doi: 10.1126/science.1321497. [DOI] [PubMed] [Google Scholar]
  38. Nishimura K. Y., Isseroff R. R., Nuccitelli R. Human keratinocytes migrate to the negative pole in direct current electric fields comparable to those measured in mammalian wounds. J Cell Sci. 1996 Jan;109(Pt 1):199–207. doi: 10.1242/jcs.109.1.199. [DOI] [PubMed] [Google Scholar]
  39. Onuma E. K., Hui S. W. Electric field-directed cell shape changes, displacement, and cytoskeletal reorganization are calcium dependent. J Cell Biol. 1988 Jun;106(6):2067–2075. doi: 10.1083/jcb.106.6.2067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Parekh A. B., Terlau H., Stühmer W. Depletion of InsP3 stores activates a Ca2+ and K+ current by means of a phosphatase and a diffusible messenger. Nature. 1993 Aug 26;364(6440):814–818. doi: 10.1038/364814a0. [DOI] [PubMed] [Google Scholar]
  41. Penner R., Matthews G., Neher E. Regulation of calcium influx by second messengers in rat mast cells. Nature. 1988 Aug 11;334(6182):499–504. doi: 10.1038/334499a0. [DOI] [PubMed] [Google Scholar]
  42. Poo M. In situ electrophoresis of membrane components. Annu Rev Biophys Bioeng. 1981;10:245–276. doi: 10.1146/annurev.bb.10.060181.001333. [DOI] [PubMed] [Google Scholar]
  43. Prausnitz M. R., Milano C. D., Gimm J. A., Langer R., Weaver J. C. Quantitative study of molecular transport due to electroporation: uptake of bovine serum albumin by erythrocyte ghosts. Biophys J. 1994 May;66(5):1522–1530. doi: 10.1016/S0006-3495(94)80943-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Putney J. W., Jr Capacitative calcium entry revisited. Cell Calcium. 1990 Nov-Dec;11(10):611–624. doi: 10.1016/0143-4160(90)90016-n. [DOI] [PubMed] [Google Scholar]
  45. Randriamampita C., Tsien R. Y. Emptying of intracellular Ca2+ stores releases a novel small messenger that stimulates Ca2+ influx. Nature. 1993 Aug 26;364(6440):809–814. doi: 10.1038/364809a0. [DOI] [PubMed] [Google Scholar]
  46. Robinson K. R. The responses of cells to electrical fields: a review. J Cell Biol. 1985 Dec;101(6):2023–2027. doi: 10.1083/jcb.101.6.2023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Stossel T. P. From signal to pseudopod. How cells control cytoplasmic actin assembly. J Biol Chem. 1989 Nov 5;264(31):18261–18264. [PubMed] [Google Scholar]
  48. Thastrup O., Cullen P. J., Drøbak B. K., Hanley M. R., Dawson A. P. Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2(+)-ATPase. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2466–2470. doi: 10.1073/pnas.87.7.2466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Tsien R. W., Tsien R. Y. Calcium channels, stores, and oscillations. Annu Rev Cell Biol. 1990;6:715–760. doi: 10.1146/annurev.cb.06.110190.003435. [DOI] [PubMed] [Google Scholar]
  50. Weeds A. Actin-binding proteins--regulators of cell architecture and motility. Nature. 1982 Apr 29;296(5860):811–816. doi: 10.1038/296811a0. [DOI] [PubMed] [Google Scholar]
  51. Williams R. M., Piston D. W., Webb W. W. Two-photon molecular excitation provides intrinsic 3-dimensional resolution for laser-based microscopy and microphotochemistry. FASEB J. 1994 Aug;8(11):804–813. doi: 10.1096/fasebj.8.11.8070629. [DOI] [PubMed] [Google Scholar]
  52. Zimmermann U. Electric field-mediated fusion and related electrical phenomena. Biochim Biophys Acta. 1982 Nov 30;694(3):227–277. doi: 10.1016/0304-4157(82)90007-7. [DOI] [PubMed] [Google Scholar]
  53. von Tscharner V., Prod'hom B., Baggiolini M., Reuter H. Ion channels in human neutrophils activated by a rise in free cytosolic calcium concentration. 1986 Nov 27-Dec 3Nature. 324(6095):369–372. doi: 10.1038/324369a0. [DOI] [PubMed] [Google Scholar]

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