Rojas Robert, Segovia Christopher, Trombert Annette Nicole, Santander Javier, Manque Patricio
Nucleus for Microbiology and Immunity, Center for Genomics and Bioinformatics, Faculty of Sciences, Universidad Mayor, Campus Huechuraba, Camino La Pirámide 5750, Huechuraba, Santiago, Chile,
Curr Microbiol. 2014 Oct;69(4):541-8. doi: 10.1007/s00284-014-0620-x. Epub 2014 Jun 4.
Crithidia fasciculata represents a very interesting model organism to study biochemical, cellular, and genetic processes unique to members of the family of the Trypanosomatidae. Thus, C. fasciculata parasitizes several species of insects and has been widely used to test new therapeutic strategies against parasitic infections. By using tunicamycin, a potent inhibitor of glycosylation in asparaginyl residues of glycoproteins (N-glycosylation), we demonstrate that N-glycosylation in C. fasciculata cells is involved in modulating glucose uptake, dramatically impacting growth, and cell adhesion. C. fasciculata treated with tunicamycin was severely affected in their ability to replicate and to adhere to polystyrene substrates and losing their ability to aggregate into small and large groups. Moreover, under tunicamycin treatment, the parasites were considerably shorter and rounder and displayed alterations in cytoplasmic vesicles formation. Furthermore, glucose uptake was significantly impaired in a tunicamycin dose-dependent manner; however, no cytotoxic effect was observed. Interestingly, this effect was reversible. Thus, when tunicamycin was removed from the culture media, the parasites recovered its growth rate, cell adhesion properties, and glucose uptake. Collectively, these results suggest that changes in the tunicamycin-dependent glycosylation levels can influence glucose uptake, cell growth, and adhesion in the protozoan parasite C. fasciculata.
fasciculata锥虫是一种非常有趣的模式生物，可用于研究锥虫科成员特有的生化、细胞和遗传过程。因此， fasciculata锥虫寄生于多种昆虫，已被广泛用于测试针对寄生虫感染的新治疗策略。通过使用衣霉素（一种糖蛋白天冬酰胺残基糖基化的有效抑制剂，即N-糖基化），我们证明fasciculata锥虫细胞中的N-糖基化参与调节葡萄糖摄取，对生长和细胞黏附产生显著影响。用衣霉素处理的fasciculata锥虫在复制和黏附于聚苯乙烯底物的能力上受到严重影响，并失去聚集形成小群体和大群体的能力。此外，在衣霉素处理下，寄生虫明显更短更圆，并且细胞质囊泡形成出现改变。此外，葡萄糖摄取以衣霉素剂量依赖性方式显著受损；然而，未观察到细胞毒性作用。有趣的是，这种作用是可逆的。因此，当从培养基中去除衣霉素时，寄生虫恢复其生长速率、细胞黏附特性和葡萄糖摄取。总的来说，这些结果表明，依赖衣霉素的糖基化水平变化可影响原生动物寄生虫fasciculata锥虫的葡萄糖摄取、细胞生长和黏附。
