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  • Title: Effects of aphidicolin and hydroxyurea on the cell cycle and differentiation of Trypanosoma brucei bloodstream forms.
    Author: Mutomba MC, Wang CC.
    Journal: Mol Biochem Parasitol; 1996 Sep; 80(1):89-102. PubMed ID: 8885225.
    Abstract:
    The effects of aphidicolin (APH) and hydroxyurea (HU) on the cell cycle and differentiation of Trypanosoma brucei bloodstream forms were studied. APH (0.1 microgram ml-1) inhibited cell division, but did not inhibit DNA synthesis. Most of the cells were arrested in the G2 phase of the cell cycle, with each cell containing two kinetoplasts, but only one nucleus. Recovery of the arrested cells showed a 24-h lag period compared to controls. Higher concentrations of APH (1 and 10 micrograms ml-1) were required to inhibit DNA synthesis, but the cells failed to resume growth after removal of the drug. Incubation of cells with HU (7.5 micrograms ml-1) did not inhibit DNA synthesis, but arrested cells after duplicating both the kinetoplast and the nucleus. Recovery from drug arrest also showed a 24-h lag period. We therefore conclude that neither APH nor HU arrests T. brucei at the G1/S phase boundary as anticipated. The mechanisms of cell cycle arrest by APH and HU are not through inhibition of DNA synthesis, but rather through unidentified pathways, leading to growth arrest prior to nuclear division and cytokinesis respectively. Since the arrested cells do not resume normal development immediately following drug removal, APH and HU should be regarded as unsuitable agents for synchronizing T. brucei bloodstream forms. T. brucei bloodstream forms arrested with either APH or HU differentiated normally into procyclic forms in vitro, indicating that a cycle of cell division is not required for initiation of differentiation, and that the process can be initiated and completed when cells are arrested at the G2/M and M/G1 phase boundaries.
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