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The spatial organization and extraction of the wall-forming bodies of Eimeria maxima

Published online by Cambridge University Press:  14 March 2013

SONJA FRÖLICH
Affiliation:
The iThree Institute, School of Medical and Molecular Biosciences, University of Technology Sydney, PO Box 123, Broadway, Sydney, New South Wales, 2007, Australia
MICHAEL JOHNSON
Affiliation:
The iThree Institute, School of Medical and Molecular Biosciences, University of Technology Sydney, PO Box 123, Broadway, Sydney, New South Wales, 2007, Australia
MICHELLE ROBINSON
Affiliation:
The iThree Institute, School of Medical and Molecular Biosciences, University of Technology Sydney, PO Box 123, Broadway, Sydney, New South Wales, 2007, Australia
ROLF ENTZEROTH
Affiliation:
Institute of Zoology, Technische Universität Dresden, Helmholtzstraße 10, 01062 Dresden, Germany
MICHAEL WALLACH*
Affiliation:
The iThree Institute, School of Medical and Molecular Biosciences, University of Technology Sydney, PO Box 123, Broadway, Sydney, New South Wales, 2007, Australia
*
*Corresponding author: The iThree Institute, School of Medical and Molecular Biosciences, University of Technology Sydney, PO Box 123, Broadway, Sydney, New South Wales, 2007, Australia. Tel.: +61-2-9514-4082. Fax: +61-2-9514-4026. E-mail: Michael.Wallach@uts.edu.au

Summary

Eimeria maxima has been used as a model apicomplexan parasite to study sexual stage development and oocyst wall formation. A complete understanding of the wall's biochemical and biophysical properties is of great interest in research on all apicomplexan parasites. Purified gametocytes, zygotes and oocysts were analysed by three-dimensional confocal microscopy, and wide-field fluorescent microscopy was used to investigate the appearance and spatial organization of the 2 types of wall-forming bodies (WFBs). In addition, a variety of staining procedures and immunoassays were used to assess the biosynthesis, metabolic activity, intactness and molecular composition of the WFBs in situ. WFBs were extracted from gametocytes/zygotes and their composition was assessed by microscopy and SDS-PAGE analysis. It was concluded that isolated gametocytes are intact and metabolically active. Additionally, it was observed that the Type 1 WFBs are aligned at the periphery of the parasite and fuse together producing neutral lipid rich patches that appear to be inserted into the space between 2 parasite-specific membranes. Finally, it was shown that the WFBs extracted from purified gametocytes had the same shape, size and staining properties as those observed in situ, and contained the major glycoprotein antigens known to be present in these organelles.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2013 

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