Metabolism and life cycle of brucei trypanosomes.

我们先前发现甘露糖在饲养层细胞支持的培养中抑制冈比亚锥虫的血型(BF)生长。为了研究高炉的代谢，有必要建立无饲养层的培养体系。用含10%胎牛血清的MEM和L-15培养液1：1混合作为完全培养液，即可完全解决上述问题。在测定高炉对甘露糖摄取时，研华GB140是测定甘露糖摄取的最佳过滤器，并用14&gt；C-甘露糖来研究这种作用方式。虽然实验应该在没有葡萄糖的情况下进行，因为已经报道了葡萄糖与甘露糖的竞争运输，但葡萄糖是高炉的主要能量来源。当BF在37ﾟC的无葡萄糖厄尔氏培养基中孵育时，BF转为静止，并通过添加糖或甘油恢复。葡萄糖和甘油有类似的作用，而甘露糖较少，半乳糖没有影响。我们的结果与布鲁氏结核杆菌通过糖酵解途径利用半乳糖的报道不一致。甘油可以作为葡萄糖的能源，不影响甘露糖的吸收。添加比热甘露糖浓度高2500倍的葡萄糖可抑制50%的甘露糖摄取。如果其他人报道的结核竞争转运是真的，相同的浓度可能足以达到50%的抑制。这些差异表明，尽管TG被认为是源于TB的亚种，但它们在生理特征上可能有很大的不同。糖酵解的中间代谢物1，6-二磷酸果糖在比甘露糖低得多的浓度下对高炉有害。原环类是与高炉不同的生命周期阶段，对这种处理具有抵抗力。在电子显微镜下，BF在加入甘露糖后2小时即可观察到胞浆和糖体的降解。

We had previously found that mannose inhibited the growth of blood form (BF) of Trypanosoma b. gambiense (Tg) in the culture supported with feeder layer cells. It is necessary to develop the cluture system without feeder layer to study the metabolism of BF. The problem was cleared by using 1 : 1 mixture of MEM and L-15 medium containing 10% fetal bovine serum as the complete medium. ADVANTEC GB140 was the best filter to harmest BF in determination of mannose uptake with ^<14>C-mannose to invesigate that mode of action. Though the experiment shouldbe performed in the absence of glucose as competitive transport with mannose had been reported, glucose is an eminent energy source of BF. BF turned inmobile when incubated in glucose-free Earle's BSS at 37ﾟC the restored by addition of sugar or glycerol. Glucose and glycerol had a similar, while mannose less and galactorse no effect. Our result did not accord with the report that T. b. Brucei (Tb) utilize galactose through glycolytic pathway. Glycerol could be energy source for glucose with no influence on mannose uptake. ^<14>C-mannose uptake was inhibited 50% by addition of 2, 500 times higher concentration of glucose than that of hot mannose. If the competitive transport of Tb reported by others was true, equal concentration might be enough for 50% inhibition. Those discrepancies suggest taht Tg, Howerer, is accepted to be subspecies derived from Tb, they might be quite different in physiological features each other. Fructose 1, 6-bisphosphate, the intermediate metabolite in glycolysis, unexpectedly did harm on BF at much lower concentration than mannose did. Procyclic, a different life cycle stage from BF, resisted to such treatment. In electron micrographs degradation in cytoplasm and Glycosome of BF was observed as early as 2 h after addition of mannose.