日本血吸虫酚氧化酶的组化定位研究

STUDIES ON HISTOCHEMICAL LOCALIZATION OF PHENOL OXIDASE IN SCHISTOSOMA JAPONICUM

目的 观察酚氧化酶 (phenol oxidase,PO)在日本血吸虫成虫的组织学分布。方法 酚氧化酶组化方法 :将 4 2 d虫龄的日本血吸虫活成虫置含 2 5 mm ol/ L 邻苯二酚的磷酸盐缓冲液 (p H6 .8)中 ,37℃孵育 30 m in后 ,转移虫体至载玻片上 ,吸去虫体表面液体 ,将其中一部分虫体置Olym pus显微镜下观察酚氧化酶在虫体的组织学分布。荧光组织化学方法 :经上述过程处理后 ,在另一部分虫体上滴加 2滴含 0 .0 5 %戊巴比妥钠的 PBS溶液 ,然后置 L eica荧光显微镜下观察酚氧化酶在虫体的组织学分布。结果 酶组织化学方法显示 ,日本血吸虫酚氧化酶仅分布于雌虫卵黄腺及子宫内虫卵卵壳表面 ,呈现棕褐色显色反应 ;雌虫卵巢和雄虫均未发现酚氧化酶活性。然而 ,荧光组织化学方法显示 ,酚氧化酶除主要分布于雌虫卵黄腺及子宫内虫卵卵壳表面 ,呈现强荧光外 ,还少量分布于雄虫体壁表层 ,呈现弱荧光反应。结论 不仅日本血吸虫雌虫含有酚氧化酶活性 ,而且雄虫也含酚氧化酶活性 ,只不过其含量少、活性低。荧光组织化学方法能更灵敏地显示日本血吸虫酚氧化酶活性 ,更适用于日本血吸虫酚氧化酶的组织学定位。

Objective To observe the histological distribution of phenol oxidase (PO) in Schistosoma japonicum.Methods Enzyme histochemical method: living female and male worms aged 42 d were incubated in pH 6.8 phosphate buffer containing 25 mM catechol at 37℃ for 30 minutes. Then some of those worms were observed for the histological distribution of their phenol oxidase under the Olympus microscope. Fluorescent histochemical method: after the process described above,other worms were given two drips of PBS containing 0.05% sodium phenobarbital,then were observed for the histological distribution of PO under the fluorescent microscope. Results PO of Schistosoma japonicum only located in the mature vitelline glands and on the eggshell surface of intrauterine eggs in mature female worms by the enzyme histochemical method. However,by the fluorescent histochemical method,PO was not only located in the mature vitelline glands and on the eggshell surface of intrauterine eggs in mature female worms,but also distributed throughout the body wall of male worms with a very weak activity. No PO activity was found in the ovaries of female worms by above two methods. Conclusion PO activity of S.japonicum could be detected both in female and male worms. The fluorescent histochemical method is more sensitive than the enzyme histochemical method in detecting PO activity of S.japonicum,So it is a better method for studying the histological localization of S. japonicum PO.