Hostname: page-component-78c5997874-fbnjt Total loading time: 0 Render date: 2024-11-05T03:54:37.574Z Has data issue: false hasContentIssue false

A rational approach to the serial culture of malaria parasites: evidence for a deficiency in RNA synthesis during the first cycle in vitro

Published online by Cambridge University Press:  06 April 2009

P. I. Trigg
Affiliation:
Division of Parasitology, National Institute for Medical Research, Mill Hill, London NW7 1AA
W. E. Gutteridge
Affiliation:
Division of Parasitology, National Institute for Medical Research, Mill Hill, London NW7 1AA

Extract

When ring-stage parasites are grown to the late trophozoite stage in vitro, the incorporation of [3H]adenosine into parasite DNA and RNA is in the ratio of ~ 1:1·5 and not 1:4 as expected from direct biochemical analysis of the parasite. A method is described by which large quantities of infected blood can be grown in vitro from the ring to the trophozoite stage thus allowing direct biochemical analysis of parasites grown in vitro. The results of direct biochemical analysis indicated that parasites grown in vitro have similar DNA and protein contents to those grown in vitro but that RNA contents are much less in vitro. Quantitative histochemical studies also indicated a deficiency in RNA in parasites grown in vitro. It is concluded from the evidence of three distinct methods that RNA synthesis is defective during the first asexual cycle in vitro. In the second cycle all three macromolecular biosyntheses studied are reduced when compared to the first so that the actual rate of RNA synthesis is further reduced when compared to that in vitro. It is suggested that the successive decrease in RNA synthesis parallels the reduction in multiplication obtained on subculture of the parasite.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1972

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

Ball, E. G., McKee, R. W., Anfinsen, C. B., Cruz, W. O., & Geiman, Q. M., (1948). Studies on malarial parasites IX. Chemical and metabolic changes during growth in vivo and in vitro. Journal of Biological Chemistry 175, 547–71.CrossRefGoogle Scholar
Bray, G. A., (1960). A simple efficient liquid scintillator for counting aqueous solutions in a liquid scintillation counter. Analytical Biochemistry 1, 279–85.CrossRefGoogle Scholar
Brown, I. N., Brown, K. N., & Hills, L. A., (1966). The separation of Plasmodium knowlesi from host cells. Transactions of the Royal Society of Tropical Medicine and Hygiene 60, 3.CrossRefGoogle Scholar
Burton, K., (1960). A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. The Biochemical Journal 62, 315–23.CrossRefGoogle Scholar
Deitch, A. D., (1964). A method for the cytophotometric estimation of nucleic acids using methylene blue. The Journal of Histochemistry and Cytochemistry 12, 451–61.CrossRefGoogle ScholarPubMed
Gale, E. F., & Folkes, J. P., (1953). The assimilation of amino acids by bacteria 14. Nucleic acid and protein synthesis in Staphylococcus aureus. The Biochemical Journal 53, 483–92.CrossRefGoogle Scholar
Geiman, Q. M., Siddiqui, W. A., & Schnell, J. V., (1966). In vitro studies on erythrocytic stages of Plasmodia: medium improvement and results with several species of malaria parasite. Military Medicine, 131 (Suppl.), 1015–25.CrossRefGoogle Scholar
Gutteridge, W. E., (1969). Some effects of pentamidine di-isethionate on Crithidia fasciculata. The Journal of Protozoology 16, 306–11.CrossRefGoogle ScholarPubMed
Gutteridge, W. E., & Trigg, P. I., (1970). Incorporation of radioactive precursors into DNA and RNA of Plasmodium knowlesi in vitro. The Journal of Protozoology 17, 8996.CrossRefGoogle ScholarPubMed
Gutteridge, W. E., & Trigg, P. I., (1971). Properties of DNA from some malarial parasites. Parasitology 62, 209–19.CrossRefGoogle ScholarPubMed
Krebs, H. A., & Eggleston, L. V., (1940). The oxidation of pyruvate in pigeon breast muscle. The Biochemical Journal 34, 442–59.CrossRefGoogle ScholarPubMed
Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J., (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 265–75.CrossRefGoogle ScholarPubMed
Mejbaum, W., (1939). Über die bestimmung kleiner pentosemengen, insbesondere in derivaten der adenylsäure. Hoppe-Seyler's Zeitschrift für Physiologische Chemie 258, 117–20.CrossRefGoogle Scholar
Trigg, P. I., (1968). A new continuous perfusion technique for the cultivation of malaria parasites in vitro. Transactions of the Royal Society of Tropical Medicine and Hygiene 62, 371–8.CrossRefGoogle Scholar
Trigg, P. I., (1969 a). The use of proprietary tissue culture media for the cultivation in vitro of the erythrocytic stages of Plasmodium knowlesi. Parasitology 59, 925–35.CrossRefGoogle ScholarPubMed
Trigg, P. I., (1969 b). Some factors affecting the cultivation in vitro of the erythrocytic stages of Plasmodium knowlesi. Parasitology 59, 915–24.CrossRefGoogle ScholarPubMed
Trigg, P. I., & Gutteridge, W. E., (1971). A minimal medium for the growth of Plasmodium knowlesi in dilution cultures. Parasitology 62, 113–23.CrossRefGoogle ScholarPubMed
Trigg, P. I., Gutteridge, W. E., & Williamson, J., (1971). The effects of cordycepin on malaria parasites. Transactions of the Royal Society of Tropical Medicine and Hygiene 65, 514–20.CrossRefGoogle ScholarPubMed
Warhurst, D. C., & Williamson, J., (1970). Ribonucleic acid from Plasmodium knowlesi before and after chloroquine treatment. Chemical-Biological Interactions 2, 89106.CrossRefGoogle ScholarPubMed