Metabolic differentiation of bloodstream forms of Trypanosoma brucei brucei into procyclic forms in hemin-depleted medium and in the presence of respiratory inhibitors

Bloodforms of Trypanosoma brucei brucei (STIB 247) differentiated in vitro into procyclic forms as described in the accompanying paper (Markos et al. 1989). The importance of the respiratory chain for the process was tested by the inhibition of its development (omission of hemin from the medium) or function (respiratory inhibitors). In the absence of hemin, all enzyme markers of the procyclic state, except for hemoproteins, developed to 50-70 per cent of control values. The presence of hemin is therefore not essential for the onset of differentiation, although the process cannot be completed under hemin limitation. Addition of 1 mumole.dm-3 KCN, 10 mumole.dm-3 antimycin A, or 100 mumole.dm-3 salicyl hydroxamate (SHAM) did not block the differentiation, although it proceeded at a slower rate. The development of the inner mitochondrial membrane markers--succinate: cytochrome c reductase, and NADH: cytochrome c reductase--was strongly inhibited by KCN or antimycin. None of these inhibitors had a significant effect on the activity of procyclic state marker--glycosomal malate dehydrogenase.