Apfel H, Eisenbeiss W F, Meyer T F
Max-Planck-Institut für Biologie, Abteilung Infektionsbiologie, Tübingen, Germany.
Mol Biochem Parasitol. 1992 May;52(1):63-73. doi: 10.1016/0166-6851(92)90036-j.
This study describes the dynamics and the biochemical nature of changes in the surface of the filarial nematode Acanthocheilonema viteae after its transmission into the vertebrate host. Vector-derived third-stage larvae (mL3) were inoculated into naive Meriones unguiculatus and recovered from the tissues at different times post-infection until their moult to fourth-stage larvae (L4). Surface-specific labelling with fluoresceinated lectins revealed that the larvae are covered by a carbohydrate envelope. Although the mL3 envelope was strongly reduced one day after transmission, new surface carbohydrates appeared until the onset of moulting, some of which could also be identified on the surface of L4. In general, surface carbohydrates were partially shed by moving larvae, suggesting a loose association of these components in the epicuticle. The fate of cuticular lipids and proteins of L3 and L4 was monitored by external 125I-labelling and differential extraction of the components. Thin-layer chromatography of surface-labelled lipids revealed only minor changes 1 day after parasite transmission. Afterwards the number of lipids accessible to label decreased further until moulting was complete. Two-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis of surface-labelled proteins showed a consistent surface exposure of mL3 specific proteins until 1 day post-infection. Thereafter, the composition of surface-labelled proteins changed rapidly, resembling that of the L4 as early as several days before moulting. During this period individual differences in the composition of surface proteins were evident.
本研究描述了丝状线虫旋盘尾丝虫传播至脊椎动物宿主后,其体表变化的动态过程及生化特性。将媒介传播的三期幼虫(mL3)接种到未感染过的长爪沙鼠体内,并在感染后的不同时间从组织中获取幼虫,直至其蜕皮为四期幼虫(L4)。用荧光素标记的凝集素进行表面特异性标记显示,幼虫被一层碳水化合物包膜所覆盖。尽管在传播后一天mL3的包膜显著减少,但新的表面碳水化合物在蜕皮开始前持续出现,其中一些也能在L4的表面被识别。总体而言,移动的幼虫会部分脱落表面碳水化合物,表明这些成分在表皮层中结合松散。通过外部¹²⁵I标记和成分的差异提取来监测L3和L4的表皮脂质和蛋白质的命运。表面标记脂质的薄层层析显示,寄生虫传播后1天仅有微小变化。此后,可被标记的脂质数量进一步减少,直至蜕皮完成。表面标记蛋白质的二维十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳显示,mL3特异性蛋白质在感染后1天一直持续在表面暴露。此后,表面标记蛋白质的组成迅速变化,早在蜕皮前几天就类似于L4的组成。在此期间,表面蛋白质组成的个体差异明显。