Wiltshire C J, Sutherland S K, Fenner P J, Young A R
Department of Pharmacology, University of Melbourne, Victoria, Australia.
Wilderness Environ Med. 2000 Winter;11(4):241-50. doi: 10.1580/1080-6032(2000)011[0241:oapcov]2.3.co;2.
To optimize venom extraction and to undertake preliminary biochemical studies of venom from the box jellyfish (Chironex fleckeri), the Irukandji jellyfish (Carukia barnesi), and the blubber jellyfish (Catostylus mosaicus).
Lyophilized crude venoms from box jellyfish tentacles and whole Irukandji jellyfish were prepared in water by homogenization, sonication, and rapid freeze thawing. A second technique, consisting of grinding samples with a glass mortar and pestle and using phosphate-buffered saline, was used to prepare crude venom from isolated nematocysts of the box jellyfish, the bells of Irukandji jellyfish, and the oral lobes of blubber jellyfish. Venoms were compared by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot test. Toxicity of some venoms was determined by intravenous median lethal dose assay in mice.
Different venom extraction techniques produced significantly different crude venoms for both box and Irukandji jellyfish. Irukandji and blubber venom SDS-PAGE protein profiles were established for the first time. Analysis of Western blot tests revealed that box jellyfish antivenin reacted specifically with the venom of each jellyfish. Toxicity was found in Irukandji jellyfish venom derived by use of the mortar-and-pestle method, but not in the lyophilized venom.
Glass mortar-and-pestle grinding and use of an appropriate buffer was found to be a simple and suitable method for the preparation of venom from each jellyfish species studied. This study contributes to biochemical investigations of jellyfish venoms, particularly the venom of the Irukandji jellyfish, for which there are, to our knowledge, no published studies. It also highlights the importance of optimizing venom extraction as the first step toward understanding the complex biological effects of jellyfish venoms.
优化三种箱形水母(曳手水母)、伊鲁坎吉水母和绵水母的毒液提取方法,并对其毒液进行初步生化研究。
通过匀浆、超声处理和快速冻融,将箱形水母触手及完整的伊鲁坎吉水母的冻干粗毒液用水制备。第二种方法是用玻璃研钵和研杵研磨样品并使用磷酸盐缓冲盐水,用于从箱形水母的分离刺丝囊、伊鲁坎吉水母的伞部及绵水母的口叶制备粗毒液。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹试验对毒液进行比较。部分毒液的毒性通过小鼠静脉注射半数致死剂量测定法确定。
不同的毒液提取技术对箱形水母和伊鲁坎吉水母产生了显著不同的粗毒液。首次建立了伊鲁坎吉水母和绵水母毒液的SDS-PAGE蛋白质图谱。蛋白质免疫印迹试验分析表明,箱形水母抗蛇毒素与每种水母的毒液发生特异性反应。用研钵-研杵法获得的伊鲁坎吉水母毒液具有毒性,而冻干毒液则无毒性。
发现用玻璃研钵和研杵研磨并使用合适的缓冲液是从所研究的每种水母制备毒液的简单且合适的方法。本研究有助于水母毒液的生化研究,特别是伊鲁坎吉水母毒液,据我们所知,尚无关于该毒液的已发表研究。该研究还强调了优化毒液提取作为理解水母毒液复杂生物学效应第一步的重要性。