Schindler J F, de Vries U
Institut für Anatomie, Universität Regensburg, Federal Republic of Germany.
Cell Tissue Res. 1988 Jul;253(1):115-28. doi: 10.1007/BF00221746.
The endodermal trophotaenial epithelium in goodeid embryos acts as a placental exchange site. Fine structural and cytochemical data indicate that the trophotaenial absorptive cells are endocytotically highly active. To test their micropinocytotic capacity and characterize the cellular mechanisms involved in membrane, solute and ligand movements, living embryos of Xenoophorus captivus were incubated in saline media containing horseradish peroxidase (HRP) and/or cationized ferritin (CF) in vitro, and the uptake of these tracer proteins examined by both time sequence analysis and pulse-chase procedures. In some embryos, the effects of prolonged exposure to CF injected into the ovarian cavity, was also investigated. Labelling of the free cell surface was detectable with CF only, but interiorization of both probes was quick from all incubation media. Adsorptive pinocytosis of CF and fluid-phase uptake of HRP sequentially labelled pinocytic vesicles, endosomes, and lysosome-like bodies. In addition, CF-molecules were sequestered within apical tubules and small vesicles. HRP was largely excluded from both organelles and ended up in the lysosomal compartment. For CF, two alternative pathways were indicated by the pulse-chase experiments; transcellular passage and regurgitation of tracer molecules to the apical cell surface. The latter procedure involves membrane and receptor recycling, in which apical tubules are thought to mediate. In double-tracer experiments, using an 8:1 excess of HRP, external labelling with CF was light or lacking after 1-3 min, and the initial uptake-phase produced pinocytic vesicles and endosomes that mainly contained HRP-reaction product. Prolonged incubation, however, resulted in densely CF-labelled plasmalemmal invaginations and pinocytic vesicles that predominantly carried ferritin granules. After 60 min, the vacuoles of the endosomal compartment contained either high concentrations of HRP-reaction product, both tracers side by side, or virtually exclusively CF.
古氏鱼胚胎中的内胚层滋养带上皮充当胎盘交换部位。精细结构和细胞化学数据表明,滋养带吸收细胞内吞作用高度活跃。为了测试它们的微胞饮能力并表征参与膜、溶质和配体转运的细胞机制,将圈养的异鳞古氏鱼活胚胎在含有辣根过氧化物酶(HRP)和/或阳离子化铁蛋白(CF)的盐溶液培养基中进行体外培养,并通过时间序列分析和脉冲追踪程序检测这些示踪蛋白的摄取情况。在一些胚胎中,还研究了长时间暴露于注入卵巢腔的CF的影响。仅用CF可检测到游离细胞表面的标记,但两种探针从所有培养基中内化都很快。CF的吸附性胞饮作用和HRP的液相摄取依次标记了胞饮小泡、内体和溶酶体样小体。此外,CF分子被隔离在顶端小管和小泡内。HRP在很大程度上被排除在这两种细胞器之外,最终进入溶酶体区室。对于CF,脉冲追踪实验表明有两条替代途径;示踪分子的跨细胞通道和反流到顶端细胞表面。后一过程涉及膜和受体循环利用,其中顶端小管被认为起介导作用。在双示踪实验中,使用过量8:1的HRP,1 - 3分钟后CF的外部标记很淡或没有,初始摄取阶段产生的胞饮小泡和内体主要含有HRP反应产物。然而,长时间孵育导致CF标记密集的质膜内陷和胞饮小泡,其中主要携带铁蛋白颗粒。60分钟后,内体区室的液泡要么含有高浓度的HRP反应产物,两种示踪剂并排存在,要么实际上只含有CF。