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Isolation of host-cell nuclei from chick intestinal cells infected with second-generation schizonts of Eimeria necatrix

Published online by Cambridge University Press:  06 April 2009

M. A. Fernando  and
J. Pasternak
M. A. Fernando
Affiliation:
Department of Pathology, Ontario Veterinary College, University of Guelph, Guelph Ontario, Canada, N2G 2W2
J. Pasternak
Affiliation:
Department of Biology, University of Waterloo, Waterloo, Ontario, Canada, N2L 3G1
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Extract

A procedure is described for the preparation of purified nuclei of chick intestinal cells infected with Eimeria necatrix. During second-generation schizogony the host-cell membrane becomes resistant to homogenization, hypotonic shock and ultrasonication. However, brief treatment with a low concentration of trypsin (0·001 ) will disrupt the cell membrane. Thereafter, the nuclei can be selectively harvested from contaminating nuclear types and schizonts by a series of centrifugations. Electron microscopy and Feulgen—DNA microspectrophotometry showed that the nuclear preparations retained their integrity with little, if any, loss of DNA. Chemical determinations confirmed that host-cell nuclear DNA synthesis was stimulated by conccidial infection and further established that quantitatively there was a coincident increase in the amount of host nuclear RNA during schizogony. It is now feasible to examine directly the effects of coccidial infection upon the host's nuclear apparatus.

Type
Research Article
Information
Parasitology , Volume 74 , Issue 1 , February 1977 , pp. 27 - 32
Copyright
Copyright © Cambridge University Press 1977

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