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Three-Dimensional Imaging of Toxoplasma gondii–Host Cell Interactions within the Parasitophorous Vacuole

Published online by Cambridge University Press:  01 October 2004

Heide Schatten  and
Hans Ris
Heide Schatten
Affiliation:
Department of Veterinary Pathobiology, University of Missouri-Columbia, 1600 East Rollins Street, Columbia, MO 65211
Hans Ris
Affiliation:
Department of Zoology, University of Wisconsin, Madison, WI 53706
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Abstract

The protozoan parasite Toxoplasma gondii is a representative of apicomplexan parasites that invades host cells through an unconventional motility mechanism. During host cell invasion it forms a specialized membrane-surrounded compartment that is called the parasitophorous vacuole. The interactions between the host cell and parasite membranes are complex and recent studies have revealed in more detail that both the host cell and the parasite membrane contribute to the formation of the parasitophorous vacuole. By using our a new specimen preparation technique that allows three-dimensional imaging of thick-sectioned internal cell structures with high-resolution, low-voltage field emission scanning electron microscopy, we were able to visualize continuous structural interactions of the host cell membrane with the parasite within the parasitophorous vacuole. Fibrous and tubular material extends from the host cell membrane and is connected to parasite membrane components. Shorter protrusions are also elaborated from the parasite. Several of these shorter fine protrusions connect to the fibrous material of the host cell membrane. The elaborate network may be used for modifications of the parasitophorous vacuole membrane that will allow utilization of nutrients from the host cell by the parisite while it is being protected from host cell attacks. The structural interactions between parasite and host cells undergo time-dependent changes, and a fission pore is the most prominent structure left connecting the parasite with the host cell. The fission pore is anchored in the host cell by thick structural components of unknown nature. The new information gained with this technique includes structural details of fibrous and tubular material that is continuous between the parasite and host cell and can be imaged in three dimensions. We present this technique as a tool to investigate more fully the complex structural interactions of the host cell and the parasite residing in the parasitophorous vacuole.

Keywords

parasitophorous vacuoletransmission electron microscopyscanning electron microscopyapicomplexan parasiteshost cell invasionrhoptries
Type
Feature Articles
Information
Microscopy and Microanalysis , Volume 10 , Issue 5 , October 2004 , pp. 580 - 585
Copyright
© 2004 Microscopy Society of America

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References

REFERENCES

Aikawa, M., Komata, Y., Asai, T., & Midorikawa, O. (1977). Transmission and scanning electron microscopy of host cell entry by Toxoplasma gondii. Am J Pathol 87, 285–296.Google Scholar
Carruthers, V.B., Giddings, O.K., & Sibley, D. (1999). Secretion of micronemal proteins is associated with toxoplasma invasion of host cells. Cell Microbiol 1, 225–235.Google Scholar
Carruthers, V.B. & Sibley, D. (1997). Sequential protein secretion from three distinct organelles of Toxoplasma gondii accompanies invasion of human fibroblasts. Eur J Cell Biol 73, 114–123.Google Scholar
Charron, A.J. & Sibley, D.L. (2002). Host cells: Mobilizable lipid resources for the intracellular parasite Toxoplasma gondii. J Cell Sci 115, 3049–3059.Google Scholar
Coppens, I. & Joiner, K.A. (2001). Parasite-host cell interactions in Toxoplasmosis: New avenues for intervention? In Expert Reviews in Molecular Medicine, pp. 1–20. Cambridge, UK: Cambridge University Press. (http://www.ermm.cbcu.cam.ac.uk)
Dobrowolski, J.M. & Sibley, L.D. (1996). Toxoplasma invasion of mammalian cells is powered by the actin cytoskeleton of the parasite. Cell 84, 933–939.Google Scholar
Dubey, J.P. (1977). Toxoplasma, Hammondia, Besnotia, Sarcocystis, and other tissue cyst-forming coccidia of man and animals. In Parasitic Protozoa III, Kreier, J.P. (Ed.), pp. 101–237. New York: Academic Press.
Klainer, A.S., Krahenbuhl, J.L., & Remington, J.S. (1973). Scanning electron microscopy of Toxoplasma gondii. J Gen Microbiol 75, 111–118.Google Scholar
Krahenbuhl, J.L. & Remington, J.S. (1982). The immunology of Toxoplasma and toxoplasmosis. In Immunology of Parasite Infections, Cohen, S. & Warren, K.S. (Eds.), pp. 356–421. Oxford: Blackwell Science.
Lingelbach, K. & Joiner, K.A. (1998). The parasitophorous vacuole membrane surrounding Plasmodium and Toxoplasma: An unusual compartment in infected cells. J Cell Sci 111, 1467–1475.Google Scholar
Mercier, C.M., Cesbron-Delauw, M.F., & Sibley, L.D. (1998). The amphipathic alpha helices of the Toxoplasma protein GRA2 mediate post-secretory membrane association. J Cell Sci 111, 2171–2180.Google Scholar
Mercier, C.M., Dubremetz, J.-F., Rauscher, B., Lecordier, L., Sibley, D.L., & Cesborn-Delauw, M.-F. (2002). Biogenesis of nanotubular network in Toxoplasma parasitophorous vacuole induced by parasite proteins. Mol Biol Cell 13, 2397–2409.Google Scholar
Mordue, D., Håkansson, S., Niesman, I., & Sibley, L.D. (1999a). Toxoplasma gondii resides in a vacuole that avoids fusion with host cell endocytic and exocytic vesicular trafficking pathways. Exp Parasitol 92, 87–99.Google Scholar
Mordue, D.G., Desai, N., Dustin, M., & Sibley, D. (1999b). Invasion by Toxoplasma gondii establishes a moving junction that selectively excludes host cell plasma membrane proteins on the basis of their membrane anchoring. J Exp Med 190, 1783–1792.Google Scholar
Morisaki, J.H., Heuser, J.E., & Sibley, L.D. (1995). Invasion of Toxoplasma gondii by active penetration of the host cell. J Cell Sci 108, 2457–2464.Google Scholar
Nichols, B.A., Chiappino, M.L., & O'Connor, G.R. (1983). Secretion from the rhoptries of Toxoplasma gondii during host-cell invasion. J Ultrastr Res 83, 85–98.Google Scholar
Nichols, B.A. & O'Connor, R.G. (1981). Penetration of mouse peritoneal macrophages by the protozoan Toxoplasma gondii. Lab Invest 44, 324–335.Google Scholar
Pacheco-Soares, C. & DeSouza, W. (2000). Labeled probes inserted in the macrophage membrane are transferred to the parasite surface and internalized during cell division by Toxoplasma gondii. Parasitol Res 86, 11–17.Google Scholar
Pouvele, B., Gormley, J., & Taraschi, T. (1994). Redistribution of parasite and host cell membrane components during Toxoplasma gondii invasion. Cell Struct Funct 23, 159–168.Google Scholar
Ogino, N. & Yoneda, C. (1966). The fine structure and mode of division of Toxoplasma gondii. Arch Ophthalmol 75, 218–227.Google Scholar
Ris, H. (1985). The cytoplasmic filament system in critical point dried whole mounts and plastic-embedded sections. J Cell Biol 100, 1474–1487.Google Scholar
Ris, H. (1989). Three-dimensional imaging of cell ultrastructure with high resolution low voltage SEM. Inst Phys Conf Ser No 98, 16, 657–662.Google Scholar
Ris, H. (1990). Application of low voltage, high resolution SEM in the study of complex intracellular structures. In Proc. XIIth Intl Congress for Electron Microscopy, pp. 18–19. San Francisco, CA: San Francisco Press.
Ris, H. (1991). The three-dimensional structure of the nuclear pore complex as seen by high voltage electron microscopy and high resolution low voltage scanning electron microscopy. EMSA Bull 21, 54–56.Google Scholar
Ris, H. & Malecki, M. (1993). High resolution field emission scanning electron microscope imaging of internal cell structures after Epon extraction from sections: A new approach to correlative ultrastructural and immunocytochemical studies. J Struct Biol 11, 148–157.Google Scholar
Schatten, H., Sibley, L.D., & Ris, H. (2003). Structural evidence for actin-like filaments in Toxoplasma gondii using high-resolution low-voltage field emission scanning electron microscopy. Microsc Microanal 9, 330–335.Google Scholar
Scholtyseck, E. & Piekarski, G. (1965). Elektronenmikroskopische Untersuchungen and Merozoiten von Eimerien (Eimeria perforans und E. steidae) und Toxoplasma gondii. Z Parasitenkd 26, 91–115.Google Scholar
Schwab, J.C., Beckers, C.J.M., & Joiner, K.A. (1994). The parasitophorous vacuole membrane surrounding intracellular Toxoplasma gondii functions as a molecular sieve. Proc Natl Acad Sci USA 91, 509–513.Google Scholar
Sheffield, H.G. & Melton, M.L. (1968). The fine structure and reproduction of Toxoplasma gondii. J Parasitol 54, 209–226.Google Scholar
Sibley, L.D. (1995). Invasion of vertebrate cells by Toxoplasma gondii. Trends Cell Biol 5, 129–132.Google Scholar
Sibley, L.D., Krahenbuhl, J.L., Adams, G.M.W., & Weidner, E. (1986). Toxoplasma modifies macrophage phagosomes by secretion of a vesicular network rich in surface proteins. J Cell Biol 103, 867–874.Google Scholar
Sibley, L.D., Niesman, I.R., Parmley, S.F., & Cesbron-Delauw, M.F. (1995). Regulated secretion of multi-lamellar vesicles leads to formation of a tubulo-vesicular network in host cell vacuoles occupied by Toxoplasma gondii. J Cell Sci 108, 1669–1677.Google Scholar
Sinai, A.P., Webster, P., & Joiner, K.A. (1997). Association of host cell endoplasmic reticulum and mitochondria with the Toxoplasma gondii parasitophorous vacuole membrane: A high affinity interaction. J Cell Sci 110, 2117–2128.Google Scholar
Striepen, B., Crawford, M.J., Shaw, M.K., Tilney, L.G., Seeber, F., & Roos, D.S. (2000). The plastid of Toxoplasma gondii is divided by association with the centrosomes. J Cell Biol 151, 1423–1434.Google Scholar
Sulzer, A.J., Strobel, P.L., Springer, E.L., Roth, I.L., & Callaway, C.S. (1974). A comparative electron microscopic study of the morphology of Toxoplasma gondii by freeze-etch replication and thin sectioning technic. J Protozool 21, 710–714.Google Scholar
Suss-Toby, E., Zimmerberg, J., & Ward, G.E. (1996). Toxoplasma invasion: The parasitophorous vacuole is formed from host cell plasma membrane and pinches off via a fission pore. Proc Natl Acad Sci USA 93, 8413–8418.Google Scholar
Ward, G.E., Miller, L.H., & Dvorak, J.A. (1993). The origin of parasitophorous vacuole membrane lipids in malaria-infected erythrocytes. J Cell Sci 106, 237–248.Google Scholar