Graf R, Raikhel A S, Brown M R, Lea A O, Briegel H
Cell Tissue Res. 1986;245(1):19-27. doi: 10.1007/BF00218082.
A polyclonal antibody was raised against trypsin purified from the midgut of blood-fed Aedes aegypti. Using this antibody and our modification of the peroxidase-antiperoxidase immunocytochemical reaction, strong activity was found in the lumen of the midgut at the light-microscopical level. The activity was localized mainly in the posterior part of the distensible, abdominal midgut, along the periphery of the blood bolus and within the peritrophic membrane. Immunoreactivity appeared 8 h after the blood meal and was most prominent around 24 h, coinciding with our previous spectrophotometric determinations of trypsin. At the electron-microscopical level, secretory granules, immunocytochemically labelled with anti-trypsin antibody and protein A-colloidal gold, were first detected about 12 h after the blood meal. At 18 h, the secretory pathway could be followed immunocytochemically from the formation of granules in the Golgi complex until their release by exocytosis in the midgut lumen. By 24 h, there was a reduction in secretory granules, and large lysosomes appeared. The process of secretion described for this mosquito is comparable to similar events in vertebrate secretory systems and the presence of an intracellular trypsinogen is suggested.
制备了一种针对从吸食血液的埃及伊蚊中肠纯化的胰蛋白酶的多克隆抗体。使用该抗体以及我们对过氧化物酶-抗过氧化物酶免疫细胞化学反应的改进方法，在光镜水平上发现中肠腔中有强烈活性。该活性主要定位于可扩张的腹部中肠的后部，沿着血团的周边以及围食膜内。免疫反应性在血餐后8小时出现，在24小时左右最为明显，这与我们之前对胰蛋白酶的分光光度测定结果一致。在电子显微镜水平上，用抗胰蛋白酶抗体和蛋白A-胶体金进行免疫细胞化学标记的分泌颗粒在血餐后约12小时首次被检测到。在18小时时，通过免疫细胞化学方法可以追踪分泌途径，从高尔基体中颗粒的形成到它们通过胞吐作用释放到中肠腔中。到24小时时，分泌颗粒减少，出现了大的溶酶体。描述的这种蚊子的分泌过程与脊椎动物分泌系统中的类似事件相当，并提示存在细胞内胰蛋白酶原。
