Hostname: page-component-78c5997874-v9fdk Total loading time: 0 Render date: 2024-11-15T13:43:11.132Z Has data issue: false hasContentIssue false
  • English
  • Français

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
Get access Rights & Permissions [Opens in a new window]

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

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Beyer, T. V. & Shibalova, T. A. (1974). The increase of DNA quantity in the nuclei of chicken caecal cells harbouring second generation schizonts of Eimeria tenella. Parasitologia (Leningrad) 8, 449–55. (In Russian.)Google Scholar
Boffa, L. C., Vidali, G. & Allfrey, V. G. (1976). Changes in nuclear non-histone protein composition during normal differentiation and carcinogenesis of intestinal epithelial cells. Experimental Cell Research 98, 396–410.CrossRefGoogle ScholarPubMed
Browning, R. F., Patton, W. H. & Lytle, C. F. (1976). Eimeria tenella: simulation of DNA synthesis in infected cultured animal cells. Experimental Parasitology 39, 195–203.CrossRefGoogle Scholar
Burton, K. (1956). A study of the conditions and mechanisms of the diphenylamine reaction for the colorimetris estimation of deoxyribonucleic acid. Biochemical Journal 62, 315–23.CrossRefGoogle Scholar
Chauveau, J., MoulÉ, Y. & Rouiller, C. H. (1956). Isolation of pure and unaltered liver nuclei: morphology and biochemical composition. Experimental Cell Research 11, 317–21.CrossRefGoogle ScholarPubMed
Chevaillier, P. H. & Philippe, M. (1975). Isolation and fractionation of mouse liver nuclei: In Methods in Cell Biology, Vol. 10 (ed. Prescott, D. M.). pp. 70–84. New York and London: Academic Press.Google Scholar
Ernest, M. J., Schultz, G. & Feigelson, P. (1976). RNA synthesis in isolated hen oviduct nuclei. Biochemistry 15, 824–9.CrossRefGoogle ScholarPubMed
Fernando, M. A. (1974). Fine structure of the schizonts and merozoites of Eimeria acervulina in the chicken. Journal of Parasitology 60, 149–59.CrossRefGoogle ScholarPubMed
Fernando, M. A. & Pasternak, J. (1978). Isolation of chick intestinal cells infected with second-generation schizonts of Eimeria necatrix. Parasitology 74, 19–26.CrossRefGoogle Scholar
Fernando, M. A., Pasternak, J., Barrell, R. & Stockdale, P. G. H. (1974). Induction of host nuclear DNA synthesis in coccidia-infected chicken intestinal cells. International Journal for Parasitology 4, 267–76.CrossRefGoogle ScholarPubMed
Hancock, R. (1974). Interphase chromosomal deoxyribonucleoprotein isolated as a discrete structure from cultured cells. Journal of Molecular Biology 86, 649–63.CrossRefGoogle ScholarPubMed
Muramatsu, M. (1970). Isolation of nuclei and nucleoli. In Methods in Cell Physiology, Vol. 4 (ed. Prescott, D. M.), pp. 195–230. New York and London: Academic Press.Google Scholar
Roodyn, D. B. (1972). Some methods for isolating nuclei. In Subcellular Components: Preparation and Fractionation (ed. Birnie, G. D.), pp. 15–51. London: Butterworth.Google Scholar
Ruiz-Carrillo, A., Wangh, L. J., Littau, V. C. & Allfrey, V. G. (1974). Changes in histone acetyl content and in non-histone protein composition of avian erythroid cells at different stages of masturation. Journal of Biological Chemistry 249, 7358–68.CrossRefGoogle Scholar
Spelsburg, T. C., Knowler, J. T. & Moses, H. L. (1974). Specific methods for the isolation of nuclei from chick oviduct. In Methods in Enzymology 31, 263–79. New York and London: Academic Press.Google Scholar