Modha J, Roberts M C, Kennedy M W, Kusel J R
Division of Biochemistry and Molecular Biology, University of Glasgow, UK.
Parasitology. 1997 Jan;114 ( Pt 1):71-7. doi: 10.1017/s0031182096008025.
The lateral diffusion (DL) properties of the fluorescent lipid probe 5-N (octadecanoyl) aminofluorescein (AF18) inserted into the surface of muscle-stage larvae of Trichinella spiralis were investigated by fluorescence recovery after photo-bleaching. AF18 was not free to diffuse laterally in dormant larvae, and this remained unchanged after larval activation in vitro with trypsin and bile. However, a significant increase in surface fluidity of the probe was demonstrated (%R = 74.5; DL = 11.5 x 10(-9) cm2/sec) when larvae invaded intestinal epithelial tissue following oral infection of mice. Membrane-permeant photoactivatable caged cyclic AMP was used to analyse the putative mechanism responsible for this increase in lateral diffusion in the parasite surface. Although incubation of larvae with 1-50 microM caged cAMP had no effect on surface fluidity, incubation with 100 microM caged cAMP induced a substantial increase in the lateral mobility of AF18 (%R = 64.3; DL = 8.3 x 10(-11) cm2/sec) immediately following photo-activation of the caged messenger. This induced fluidity, however, was transient and the larval surface reverted to immobility within 15 min. These observations constitute the first reported measurement of the fluid properties of the surface of intracellular parasites, the first demonstration of the parasite surface fluidity altering as a result of host cell invasion and the first indication of a mechanism underlying changes in surface fluidity in parasitic helminths.
采用光漂白后荧光恢复技术,研究了插入旋毛虫肌幼虫表面的荧光脂质探针5 - N(十八烷酰)氨基荧光素(AF18)的侧向扩散(DL)特性。AF18在休眠幼虫中不能自由地进行侧向扩散,并且在用胰蛋白酶和胆汁进行体外幼虫激活后,这种情况保持不变。然而,当小鼠经口感染后幼虫侵入肠上皮组织时,探针的表面流动性显著增加(%R = 74.5;DL = 11.5×10⁻⁹ cm²/秒)。使用可透过膜的光激活笼形环磷酸腺苷来分析导致寄生虫表面侧向扩散增加的推定机制。尽管用1 - 50 μM笼形环磷酸腺苷孵育幼虫对表面流动性没有影响,但在用100 μM笼形环磷酸腺苷孵育后,笼形信使光激活后立即诱导AF18的侧向迁移率大幅增加(%R = 64.3;DL = 8.3×10⁻¹¹ cm²/秒)。然而,这种诱导的流动性是短暂的,幼虫表面在15分钟内恢复到不流动状态。这些观察结果构成了首次报道的细胞内寄生虫表面流体特性的测量,首次证明了寄生虫表面流动性因宿主细胞入侵而改变,以及首次表明了寄生蠕虫表面流动性变化的潜在机制。