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Metabolism and life cycle of brucei trypanosomes.

布氏锥虫的代谢和生命周期。

基本信息

批准号：
02670169
负责人：
FUKUMA Toshihide
金额：
$ 1.28万
依托单位：
Kurume University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02670169/
关键词：
African trypanosomes culture carbohydrate metabolism mannose blood form life cycle glycosome

项目摘要

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.

我们以前发现甘露糖抑制血型锥虫B的生长。gambiense（Tg）在饲养层细胞支持的培养物中培养。因此，有必要建立无饲养层的培养体系来研究高炉的代谢。通过使用MEM和含10%胎牛血清的L-15培养基的1：1混合物作为完全培养基，解决了该问题。ADVANTEC GB 140是测定β-C-甘露糖摄取量时最佳的滤膜<14>，以探讨其作用方式。虽然实验应该在没有葡萄糖的情况下进行，因为已经报道了与甘露糖的竞争运输，但葡萄糖是BF的一个重要能源。BF在无葡萄糖的Earle's BSS中37 ℃孵育时变得不活动，通过添加糖或甘油恢复。葡萄糖和甘油的作用相似，而甘露糖作用较小，半乳糖作用不明显。结果与T. B.布鲁氏菌（Tb）通过糖酵解途径利用半乳糖。甘油可以作为葡萄糖的能量来源，而不影响甘露糖的摄取。加入<14>2500倍于热甘露糖浓度的葡萄糖可抑制50%的C-甘露糖摄取。如果其他人报道的Tb的竞争性转运是真实的，那么相等的浓度可能足以达到50%的抑制。这些差异表明，虽然Tg被认为是Tb的亚种，但它们在生理特征上可能存在很大差异。糖酵解的中间代谢产物1，6-二磷酸果糖在比甘露糖低得多的浓度下意外地对BF造成损害。前循环，一个不同的生活史阶段，从BF，抵制这样的处理。电镜下观察到BF的胞质和糖体在加入甘露糖后2 h即发生降解。