Erttmann K D, Gallin M Y, Eggert P, Büttner D W
Section of Molecular Biology, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany.
Trop Med Int Health. 1996 Oct;1(5):558-74. doi: 10.1111/j.1365-3156.1996.tb00081.x.
The distribution of an Onchocerca volvulus ankyrin, designated E1, was studied in different O. volvulus stages and other helminths by immunohistochemistry using rabbit antibodies raised against the recombinant E1 protein. In adult O. volvulus the protein designated E1 was localized to the extracellular clefts as well as to the cytoplasm adjacent to the cell membrane in the area of the basal labyrinth in hypodermis, intestine and uterus and to a lesser extent in oviduct and vas deferens. Neuronal cell bodies were also labelled. No labelling of the basal laminae, muscles or epithelia of ovary or testis was observed. Detection of the E1 protein was associated with embryonic development. Germ cells and early morulae showed no reaction; labelling was first seen in late morulae, corresponding to the stage of gastrulation, and increased in the following embryonic stages. In microfilariae the nerve ring and the cephalic space, which represents the anterior nerve-enriched portion of the body, were labelled. In third-stage larvae of O. volvulus labelling was associated with the hypodermis, and in those of Anisakis sp. the cytoplasm adjacent to the membrane of the excretory gland cell and the basal labyrinth of the hypodermis were labelled. Following anthelminthic treatment a disruption of the labelling pattern of the E1 protein was observed in adult O. volvulus with leakage of the protein into neighbouring areas. Damage to the worm was associated with reduction and finally loss of E1 protein labelling. No E1 protein was detected in dead adult worms, embryos or microfilariae. Labelling of the same organs was observed in 8 other Onchocerca species and in several other nematodes, but no reaction was seen in trematodes. The results indicate that the EI protein is associated with neuronal structures of O. volvulus, that its presence is developmentally regulated and that it has cross-reactive homologues in other nematodes. The results suggest that E1 is a functional protein. It may be useful for the assessment of parasite damage and death as well as in the characterization of the filarial nervous system.
利用针对重组E1蛋白产生的兔抗体，通过免疫组织化学方法研究了盘尾丝虫锚蛋白（命名为E1）在盘尾丝虫不同发育阶段及其他蠕虫中的分布情况。在成年盘尾丝虫中，命名为E1的蛋白定位于细胞外裂隙以及皮下组织、肠道和子宫基底迷路区域中与细胞膜相邻的细胞质中，在输卵管和输精管中的定位程度较低。神经元细胞体也有标记。未观察到卵巢或睾丸的基膜、肌肉或上皮有标记。E1蛋白的检测与胚胎发育相关。生殖细胞和早期桑葚胚无反应；标记最早出现在晚期桑葚胚，对应原肠胚形成阶段，并在随后的胚胎阶段增加。在微丝蚴中，神经环和头部间隙（代表身体前部富含神经的部分）有标记。在盘尾丝虫的第三期幼虫中，标记与皮下组织相关，而在异尖线虫的第三期幼虫中，排泄腺细胞膜相邻的细胞质和皮下组织的基底迷路有标记。驱虫治疗后，在成年盘尾丝虫中观察到E1蛋白标记模式的破坏，蛋白渗漏到邻近区域。蠕虫损伤与E1蛋白标记的减少及最终消失相关。在死亡的成年蠕虫、胚胎或微丝蚴中未检测到E1蛋白。在其他8种盘尾丝虫和几种其他线虫中观察到相同器官的标记，但吸虫中未观察到反应。结果表明，EI蛋白与盘尾丝虫的神经元结构相关，其存在受发育调控，且在其他线虫中有交叉反应性同源物。结果提示E1是一种功能蛋白。它可能有助于评估寄生虫的损伤和死亡以及丝虫神经系统的特征描述。
