Transformation of monomorphic Trypanosoma brucei bloodstream form trypomastigotes into procyclic forms at 37 degrees C by removing glucose from the culture medium.
African trypanosomes have been shown previously to undergo efficient transformation from bloodstream forms to procyclic (insect dwelling) forms in vitro by adding citrate and/or cis-aconitate to the culture medium and lowering incubation temperature to 27 degrees C. In this paper, it is shown that strain 427 monomorphic bloodstream forms of Trypanosoma brucei grown in axenic culture at 37 degrees C can be transformed to procyclic forms by simply replacing the glucose carbon source in the culture medium with glycerol. The removal of glucose from the medium results in the loss of the variant surface glycoprotein, the acquisition of cell surface procyclic acidic repetitive protein, the synthesis of procyclic-specific glycosylphosphatidylinositol precursors and the acquisition of substantial resistance to salicyl hydroxamic acid and glycerol within 72 h. A procyclic-specific cytoskeletal protein, known to be a marker of the late stage of transformation, is fully expressed by 96 h but full trans-sialidase activity appears only after 18-30 days. The transformation process described here is slower and less efficient than that previously described for monomorphic trypanosomes, using citrate and/or cis-aconitate and temperature shift as triggers. However, the separation of the transformation process from these stimuli is significant and the effects of glucose deprivation described here may reflect some of the events that occur in vivo in the tsetse fly midgut, where glucose levels are known to be very low.
Department of Biochemistry, University of Dundee, UK. kgmilne@dundee.ac.uk
K G
KG
Milne
A R
AR
Prescott
M A
MA
Ferguson
D000818:Animals; D015415:Biomarkers; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000032:analysis; D003470:Culture Media / Q000737:chemistry*; D004351:Drug Resistance; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D005947:Glucose; D005990:Glycerol / Q000494:pharmacology; D017261:Glycosylphosphatidylinositols / Q000032:analysis; D008018:Life Cycle Stages / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D015800:Protozoan Proteins / Q000378:metabolism*; D012457:Salicylamides / Q000494:pharmacology; D013997:Time Factors; D014344:Trypanocidal Agents / Q000494:pharmacology; D014346:Trypanosoma brucei brucei / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000378:metabolism*
D015415:Biomarkers; D018797:Cell Cycle Proteins; D003470:Culture Media; D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins; D015800:Protozoan Proteins; D012457:Salicylamides; D014344:Trypanocidal Agents; D014643:Variant Surface Glycoproteins, Trypanosoma; C053277:procyclic acidic repetitive protein, Trypanosoma; C005703:salicylhydroxamic acid; D005947:Glucose; D005990:Glycerol