Abstract
C3H/10T1/2 mouse embryo fibroblasts were stimulated by a steady electric field ranging up to 10 V/cm. Some cells elongated and aligned perpendicular to the field direction. A preferential positional shift toward the cathode was observed which was inhibited by the calcium channel blocker D-600 and the calmodulin antagonist trifluoperazine. Rhodaminephalloidin labeling of actin filaments revealed a field- induced disorganization of the stress fiber pattern, which was reduced when stimulation was conducted in calcium-depleted buffer or in buffer containing calcium antagonist CoCl2, calcium channel blocker D-600, or calmodulin antagonist trifluoperazine. Treatment with calcium ionophore A23187 had similar effects, except that the presence of D-600 did not reduce the stress fiber disruption. The calcium-sensitive photoprotein aequorin was used to monitor changes in intracellular-free calcium. Electric stimulation caused an increase of calcium to the micromolar range. This increase was inhibited by calcium-depleted buffer or by CoCl2, and was reduced by D-600. A calcium-dependent mechanism is proposed to explain the observed field-directed cell shape changes, preferential orientation, and displacement.
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- Adelstein R. S., Eisenberg E. Regulation and kinetics of the actin-myosin-ATP interaction. Annu Rev Biochem. 1980;49:921–956. doi: 10.1146/annurev.bi.49.070180.004421. [DOI] [PubMed] [Google Scholar]
- Barker A. T., Jaffe L. F., Vanable J. W., Jr The glabrous epidermis of cavies contains a powerful battery. Am J Physiol. 1982 Mar;242(3):R358–R366. doi: 10.1152/ajpregu.1982.242.3.R358. [DOI] [PubMed] [Google Scholar]
- Bereiter-Hahn J. Architecture of tissue cells. The structural basis which determines shape and locomotion of cells. Acta Biotheor. 1985;34(2-4):139–148. doi: 10.1007/BF00046779. [DOI] [PubMed] [Google Scholar]
- Blinks J. R., Prendergast F. G., Allen D. G. Photoproteins as biological calcium indicators. Pharmacol Rev. 1976 Mar;28(1):1–93. [PubMed] [Google Scholar]
- Borgens R. B. What is the role of naturally produced electric current in vertebrate regeneration and healing. Int Rev Cytol. 1982;76:245–298. doi: 10.1016/s0074-7696(08)61793-3. [DOI] [PubMed] [Google Scholar]
- Burgess W. H., Watterson D. M., Van Eldik L. J. Identification of calmodulin-binding proteins in chicken embryo fibroblasts. J Cell Biol. 1984 Aug;99(2):550–557. doi: 10.1083/jcb.99.2.550. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cobbold P. H., Goyns M. H. Measurements of the free calcium concentration of single quiescent human fibroblasts before and after serum addition. Biosci Rep. 1983 Jan;3(1):79–86. doi: 10.1007/BF01121574. [DOI] [PubMed] [Google Scholar]
- Cooper M. S., Keller R. E. Perpendicular orientation and directional migration of amphibian neural crest cells in dc electrical fields. Proc Natl Acad Sci U S A. 1984 Jan;81(1):160–164. doi: 10.1073/pnas.81.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Erickson C. A., Nuccitelli R. Embryonic fibroblast motility and orientation can be influenced by physiological electric fields. J Cell Biol. 1984 Jan;98(1):296–307. doi: 10.1083/jcb.98.1.296. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ferrier J., Ross S. M., Kanehisa J., Aubin J. E. Osteoclasts and osteoblasts migrate in opposite directions in response to a constant electrical field. J Cell Physiol. 1986 Dec;129(3):283–288. doi: 10.1002/jcp.1041290303. [DOI] [PubMed] [Google Scholar]
- Gail M. H., Boone C. W. Effect of colcemid on fibroblast motility. Exp Cell Res. 1971 Mar;65(1):221–227. doi: 10.1016/s0014-4827(71)80070-8. [DOI] [PubMed] [Google Scholar]
- Heggeness M. H., Wang K., Singer S. J. Intracellular distributions of mechanochemical proteins in cultured fibroblasts. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3883–3887. doi: 10.1073/pnas.74.9.3883. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hinkle L., McCaig C. D., Robinson K. R. The direction of growth of differentiating neurones and myoblasts from frog embryos in an applied electric field. J Physiol. 1981 May;314:121–135. doi: 10.1113/jphysiol.1981.sp013695. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jaffe L. F. Electrical currents through the developing fucus egg. Proc Natl Acad Sci U S A. 1966 Oct;56(4):1102–1109. doi: 10.1073/pnas.56.4.1102. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jaffe L. F., Nuccitelli R. An ultrasensitive vibrating probe for measuring steady extracellular currents. J Cell Biol. 1974 Nov;63(2 Pt 1):614–628. doi: 10.1083/jcb.63.2.614. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Luther P. W., Peng H. B., Lin J. J. Changes in cell shape and actin distribution induced by constant electric fields. Nature. 1983 May 5;303(5912):61–64. doi: 10.1038/303061a0. [DOI] [PubMed] [Google Scholar]
- McNeil P. L., Taylor D. L. Aequorin entrapment in mammalian cells. Cell Calcium. 1985 Apr;6(1-2):83–93. doi: 10.1016/0143-4160(85)90036-3. [DOI] [PubMed] [Google Scholar]
- 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]
- Moore L., Pastan I. A calcium requirement for movement of cultured cells. J Cell Physiol. 1979 Oct;101(1):101–108. doi: 10.1002/jcp.1041010112. [DOI] [PubMed] [Google Scholar]
- Nuccitelli R., Erickson C. A. Embryonic cell motility can be guided by physiological electric fields. Exp Cell Res. 1983 Aug;147(1):195–201. doi: 10.1016/0014-4827(83)90284-7. [DOI] [PubMed] [Google Scholar]
- Onuma E. K., Hui S. W. A calcium requirement for electric field-induced cell shape changes and preferential orientation. Cell Calcium. 1985 Jun;6(3):281–292. doi: 10.1016/0143-4160(85)90012-0. [DOI] [PubMed] [Google Scholar]
- Orida N., Feldman J. D. Directional protrusive pseudopodial activity and motility in macrophages induced by extracellular electric fields. Cell Motil. 1982;2(3):243–255. doi: 10.1002/cm.970020305. [DOI] [PubMed] [Google Scholar]
- Oster G. F. On the crawling of cells. J Embryol Exp Morphol. 1984 Nov;83 (Suppl):329–364. [PubMed] [Google Scholar]
- Poo M. M., Poo W. J., Lam J. W. Lateral electrophoresis and diffusion of Concanavalin A receptors in the membrane of embryonic muscle cell. J Cell Biol. 1978 Feb;76(2):483–501. doi: 10.1083/jcb.76.2.483. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reznikoff C. A., Brankow D. W., Heidelberger C. Establishment and characterization of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. Cancer Res. 1973 Dec;33(12):3231–3238. [PubMed] [Google Scholar]
- 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]
- Strohmeier R., Bereiter-Hahn J. Control of cell shape and locomotion by external calcium. Exp Cell Res. 1984 Oct;154(2):412–420. doi: 10.1016/0014-4827(84)90165-4. [DOI] [PubMed] [Google Scholar]
- Stump R. F., Robinson K. R. Xenopus neural crest cell migration in an applied electrical field. J Cell Biol. 1983 Oct;97(4):1226–1233. doi: 10.1083/jcb.97.4.1226. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Svitkina T. M., Neyfakh A. A., Jr, Bershadsky A. D. Actin cytoskeleton of spread fibroblasts appears to assemble at the cell edges. J Cell Sci. 1986 Jun;82:235–248. doi: 10.1242/jcs.82.1.235. [DOI] [PubMed] [Google Scholar]
- Taylor D. L., Blinks J. R., Reynolds G. Contractile basis of ameboid movement. VII. Aequorin luminescence during ameboid movement, endocytosis, and capping. J Cell Biol. 1980 Aug;86(2):599–607. doi: 10.1083/jcb.86.2.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Wieland T., Miura T., Seeliger A. Analogs of phalloidin. D-Abu2-Lys7-phalloin, an F-actin binding analog, its rhodamine conjugate (RLP) a novel fluorescent F-actin-probe, and D-Ala2-Leu7-phalloin, an inert peptide. Int J Pept Protein Res. 1983 Jan;21(1):3–10. [PubMed] [Google Scholar]
- Yang W. P., Onuma E. K., Hui S. W. Response of C3H/10T1/2 fibroblasts to an external steady electric field stimulation. Reorientation, shape change, ConA receptor and intramembranous particle distribution and cytoskeleton reorganization. Exp Cell Res. 1984 Nov;155(1):92–104. doi: 10.1016/0014-4827(84)90770-5. [DOI] [PubMed] [Google Scholar]
