Table 1.
Effect of glucose deprivation on motility and morphology in bloodstream trypanosomes.
| Treatment | Conc (mM) | 15 min | 2 h | 6 h | 8 h | 12 h | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| morp | mot | morp | mot | morp | mot | morp | mot | morp | mot | ||
| Control | 25 | 100% | 100% | 100% | 100% | 100% | 100% | 100% | 100% | 100% | 100% |
| 2-deoxy-D-glucose | 0.1 | Dead | Dead | NA | NA | NA | NA | NA | NA | NA | NA |
| 1.0 | Dead | Dead | NA | NA | NA | NA | NA | NA | NA | NA | |
| 10 | Dead | Dead | NA | NA | NA | NA | NA | NA | NA | NA | |
| D-Glucose | 0.0 | Dead | Dead | NA | NA | NA | NA | NA | NA | NA | NA |
| 0.1 | Dead | Dead | NA | NA | NA | NA | NA | NA | NA | NA | |
| 1.0 | 100% | 50% | Dead | Dead | NA | NA | NA | NA | NA | NA | |
| 5.0 | 100% | 100% | 90% | 100% | 65% | 80% | 55% | 65% | 20% | 35% | |
| 10 | 100% | 100% | 100% | 100% | 100% | 100% | 80% | 90% | 50% | 50% | |
Log-phase trypanosomes were harvested, cells washed in PBS and resuspended in HMI-9 medium supplemented with 2-deoxy-D-glucose, in standard formulation HMI-9 (25 mM) or glucose free RPMI-1640 supplemented with D-glucose as indicated. All media also contained dialyzed 10% fetal bovine serum. Cells were incubated at 37°C and observed under phase contrast at intervals. Aliquots were withdrawn at various times and mounted on pre-warmed microscope slides. Normal cells were scored as a length:breadth ratio of >5:1 while rounded cells scored with a ratio of 2:1 or less. "morp"; percent of cells retaining normal morphology. Abnormalities includes rounded cells or emergence of a BigEye flagellar pocket. "mot"; motility; cells were scored as motile if there was evidence of cell movement, including flagellar twitching. NA; not analyzed.