Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1994 Mar 1;179(3):1017–1022. doi: 10.1084/jem.179.3.1017

Role in host cell invasion of Trypanosoma cruzi-induced cytosolic-free Ca2+ transients

PMCID: PMC2191425  PMID: 8113670

Abstract

Trypanosoma cruzi enters cells by a unique mechanism, distinct from phagocytosis. Invasion is facilitated by disruption of host cell actin microfilaments, and involves recruitment and fusion of host lysosomes at the site of parasite entry. These findings implied the existence of transmembrane signaling mechanisms triggered by the parasites in the host cells before invasion. Here we show that infective trypomastigotes or their isolated membranes, but not the noninfective epimastigotes, induce repetitive cytosolic-free Ca2+ transients in individual normal rat kidney fibroblasts, in a pertussis toxin-sensitive manner. Parasite entry is inhibited by buffering or depleting host cell cytosolic-free Ca2+, or by pretreatment with Ca2+ channel blockers or pertussis toxin. In contrast, invasion is enhanced by brief exposure of the host cells to cytochalasin D. These results indicate that a trypomastigote membrane factor triggers cytosolic-free Ca2+ transients in host cells through a G-protein-coupled pathway. This signaling event may promote invasion through modulation of the host cell actin cytoskeleton.

Full Text

The Full Text of this article is available as a PDF (1.7 MB).

Selected References

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

  1. Aderem A. Signal transduction and the actin cytoskeleton: the roles of MARCKS and profilin. Trends Biochem Sci. 1992 Oct;17(10):438–443. doi: 10.1016/0968-0004(92)90016-3. [DOI] [PubMed] [Google Scholar]
  2. Andrews N. W., Hong K. S., Robbins E. S., Nussenzweig V. Stage-specific surface antigens expressed during the morphogenesis of vertebrate forms of Trypanosoma cruzi. Exp Parasitol. 1987 Dec;64(3):474–484. doi: 10.1016/0014-4894(87)90062-2. [DOI] [PubMed] [Google Scholar]
  3. Berridge M. J. Inositol trisphosphate and calcium signalling. Nature. 1993 Jan 28;361(6410):315–325. doi: 10.1038/361315a0. [DOI] [PubMed] [Google Scholar]
  4. Bourne H. R., Sanders D. A., McCormick F. The GTPase superfamily: a conserved switch for diverse cell functions. Nature. 1990 Nov 8;348(6297):125–132. doi: 10.1038/348125a0. [DOI] [PubMed] [Google Scholar]
  5. Bretscher A. Microfilament structure and function in the cortical cytoskeleton. Annu Rev Cell Biol. 1991;7:337–374. doi: 10.1146/annurev.cb.07.110191.002005. [DOI] [PubMed] [Google Scholar]
  6. Cunningham I. New culture medium for maintenance of tsetse tissues and growth of trypanosomatids. J Protozool. 1977 May;24(2):325–329. doi: 10.1111/j.1550-7408.1977.tb00987.x. [DOI] [PubMed] [Google Scholar]
  7. Della Bianca V., Grzeskowiak M., Rossi F. Studies on molecular regulation of phagocytosis and activation of the NADPH oxidase in neutrophils. IgG- and C3b-mediated ingestion and associated respiratory burst independent of phospholipid turnover and Ca2+ transients. J Immunol. 1990 Feb 15;144(4):1411–1417. [PubMed] [Google Scholar]
  8. Docampo R., Moreno S. N., Vercesi A. E. Effect of thapsigargin on calcium homeostasis in Trypanosoma cruzi trypomastigotes and epimastigotes. Mol Biochem Parasitol. 1993 Jun;59(2):305–313. doi: 10.1016/0166-6851(93)90228-p. [DOI] [PubMed] [Google Scholar]
  9. Dvorak J. A., Hyde T. P. Trypanosoma cruzi: interaction with vertebrate cells in vitro. 1. Individual interactions at the cellular and subcellular levels. Exp Parasitol. 1973 Oct;34(2):268–283. doi: 10.1016/0014-4894(73)90087-8. [DOI] [PubMed] [Google Scholar]
  10. Francis C. L., Ryan T. A., Jones B. D., Smith S. J., Falkow S. Ruffles induced by Salmonella and other stimuli direct macropinocytosis of bacteria. Nature. 1993 Aug 12;364(6438):639–642. doi: 10.1038/364639a0. [DOI] [PubMed] [Google Scholar]
  11. Greenberg S., el Khoury J., di Virgilio F., Kaplan E. M., Silverstein S. C. Ca(2+)-independent F-actin assembly and disassembly during Fc receptor-mediated phagocytosis in mouse macrophages. J Cell Biol. 1991 May;113(4):757–767. doi: 10.1083/jcb.113.4.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hagiwara S., Byerly L. Calcium channel. Annu Rev Neurosci. 1981;4:69–125. doi: 10.1146/annurev.ne.04.030181.000441. [DOI] [PubMed] [Google Scholar]
  13. Hall B. F., Webster P., Ma A. K., Joiner K. A., Andrews N. W. Desialylation of lysosomal membrane glycoproteins by Trypanosoma cruzi: a role for the surface neuraminidase in facilitating parasite entry into the host cell cytoplasm. J Exp Med. 1992 Aug 1;176(2):313–325. doi: 10.1084/jem.176.2.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hescheler J., Rosenthal W., Hinsch K. D., Wulfern M., Trautwein W., Schultz G. Angiotensin II-induced stimulation of voltage-dependent Ca2+ currents in an adrenal cortical cell line. EMBO J. 1988 Mar;7(3):619–624. doi: 10.1002/j.1460-2075.1988.tb02855.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Heuser J. Changes in lysosome shape and distribution correlated with changes in cytoplasmic pH. J Cell Biol. 1989 Mar;108(3):855–864. doi: 10.1083/jcb.108.3.855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Huang A. J., Manning J. E., Bandak T. M., Ratau M. C., Hanser K. R., Silverstein S. C. Endothelial cell cytosolic free calcium regulates neutrophil migration across monolayers of endothelial cells. J Cell Biol. 1993 Mar;120(6):1371–1380. doi: 10.1083/jcb.120.6.1371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jaconi M. E., Lew D. P., Carpentier J. L., Magnusson K. E., Sjögren M., Stendahl O. Cytosolic free calcium elevation mediates the phagosome-lysosome fusion during phagocytosis in human neutrophils. J Cell Biol. 1990 May;110(5):1555–1564. doi: 10.1083/jcb.110.5.1555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kaziro Y., Itoh H., Kozasa T., Nakafuku M., Satoh T. Structure and function of signal-transducing GTP-binding proteins. Annu Rev Biochem. 1991;60:349–400. doi: 10.1146/annurev.bi.60.070191.002025. [DOI] [PubMed] [Google Scholar]
  19. Lew V. L., Garcia-Sancho J. Use of the ionophore A23187 to measure and control cytoplasmic Ca2+ levels in intact red cells. Cell Calcium. 1985 Apr;6(1-2):15–23. doi: 10.1016/0143-4160(85)90031-4. [DOI] [PubMed] [Google Scholar]
  20. Llinás R., Sugimori M., Silver R. B. Microdomains of high calcium concentration in a presynaptic terminal. Science. 1992 May 1;256(5057):677–679. doi: 10.1126/science.1350109. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Nathanson M. H., Burgstahler A. D. Subcellular distribution of cytosolic Ca2+ in isolated rat hepatocyte couplets: evaluation using confocal microscopy. Cell Calcium. 1992 Feb;13(2):89–98. doi: 10.1016/0143-4160(92)90002-a. [DOI] [PubMed] [Google Scholar]
  23. Nathanson M. H., Moyer M. S., Burgstahler A. D., O'Carroll A. M., Brownstein M. J., Lolait S. J. Mechanisms of subcellular cytosolic Ca2+ signaling evoked by stimulation of the vasopressin V1a receptor. J Biol Chem. 1992 Nov 15;267(32):23282–23289. [PubMed] [Google Scholar]
  24. Nogueira N., Cohn Z. Trypanosoma cruzi: mechanism of entry and intracellular fate in mammalian cells. J Exp Med. 1976 Jun 1;143(6):1402–1420. doi: 10.1084/jem.143.6.1402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ortega-Barria E., Pereira M. E. A novel T. cruzi heparin-binding protein promotes fibroblast adhesion and penetration of engineered bacteria and trypanosomes into mammalian cells. Cell. 1991 Oct 18;67(2):411–421. doi: 10.1016/0092-8674(91)90192-2. [DOI] [PubMed] [Google Scholar]
  26. Pace J., Hayman M. J., Galán J. E. Signal transduction and invasion of epithelial cells by S. typhimurium. Cell. 1993 Feb 26;72(4):505–514. doi: 10.1016/0092-8674(93)90070-7. [DOI] [PubMed] [Google Scholar]
  27. Schenkman S., Andrews N. W., Nussenzweig V., Robbins E. S. Trypanosoma cruzi invade a mammalian epithelial cell in a polarized manner. Cell. 1988 Oct 7;55(1):157–165. doi: 10.1016/0092-8674(88)90018-9. [DOI] [PubMed] [Google Scholar]
  28. Schenkman S., Robbins E. S., Nussenzweig V. Attachment of Trypanosoma cruzi to mammalian cells requires parasite energy, and invasion can be independent of the target cell cytoskeleton. Infect Immun. 1991 Feb;59(2):645–654. doi: 10.1128/iai.59.2.645-654.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Simon M. I., Strathmann M. P., Gautam N. Diversity of G proteins in signal transduction. Science. 1991 May 10;252(5007):802–808. doi: 10.1126/science.1902986. [DOI] [PubMed] [Google Scholar]
  30. Stossel T. P. On the crawling of animal cells. Science. 1993 May 21;260(5111):1086–1094. doi: 10.1126/science.8493552. [DOI] [PubMed] [Google Scholar]
  31. Stryer L., Bourne H. R. G proteins: a family of signal transducers. Annu Rev Cell Biol. 1986;2:391–419. doi: 10.1146/annurev.cb.02.110186.002135. [DOI] [PubMed] [Google Scholar]
  32. Tardieux I., Webster P., Ravesloot J., Boron W., Lunn J. A., Heuser J. E., Andrews N. W. Lysosome recruitment and fusion are early events required for trypanosome invasion of mammalian cells. Cell. 1992 Dec 24;71(7):1117–1130. doi: 10.1016/s0092-8674(05)80061-3. [DOI] [PubMed] [Google Scholar]
  33. Thomas P., Wong J. G., Almers W. Millisecond studies of secretion in single rat pituitary cells stimulated by flash photolysis of caged Ca2+. EMBO J. 1993 Jan;12(1):303–306. doi: 10.1002/j.1460-2075.1993.tb05657.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Trifaró J. M., Rodríguez del Castillo A., Vitale M. L. Dynamic changes in chromaffin cell cytoskeleton as prelude to exocytosis. Mol Neurobiol. 1992 Winter;6(4):339–358. doi: 10.1007/BF02757940. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES