Parracho Tiago, Morais Zilda
Centro de Investigação Interdisciplinar Egas Moniz (CiiEM), Egas Moniz Cooperativa de Ensino Superior, Quinta da Granja, Monte de Caparica 2829-511 Caparica, Portugal.
J Venom Anim Toxins Incl Trop Dis. 2015 Oct 5;21:40. doi: 10.1186/s40409-015-0037-4. eCollection 2015.
More than 200 Scyphozoa species have been described, but few have been properly studied regarding their chemical and genetic characteristics. Catostylus tagi, an edible Scyphozoa and the sole European Catostylidae, occurs in summer at Tagus and Sado estuaries. Neither a systematic comparison between the two Catostylus communities nor a chemical approach on their nematocytes had been carried out yet.
In order to achieve these purposes, optimisation of DNA extraction and of histochemical staining procedures were developed. Catostylus specimens from Tagus and Sado estuaries were compared by ribosomal 18S, 28S, and ITS1 partial sequencing. The morphochemistry of nematocytes was studied by optical and electronic microscopy.
Macroscopic and molecular results indicated that both communities belong to the same species, C. tagi. The hematoxylin and eosin staining allowed the visualisation of nematocyst genesis and indicated a basic character for the macromolecules on the shaft of euryteles and on the tubule of isorhizae and birhopaloids. By Masson's trichrome procedure, the basic properties of the tubules were confirmed and a collagenous profile for the toxins was suggested. Results of the alcian blue staining showed that the outer membrane of nematocyte may consist of macromolecules with acidic polysaccharides, consistent with NOWA and nematogalectin glycoproteins detected in Hydra, but also with poly-gamma-glutamate complex, chitin-like polysaccharides and hyaluronic acids. Through the von Kossa assays, calcium was detected; its position suggested interactions with polysaccharides of the membrane, with proteins of the contractile system or with both.
The optimisation of sample preparation for DNA extraction may facilitate further studies on little known jellyfish species. The improvement of the smear procedure simplified the use of stained reactions in zooplankton. Moreover, it was shown that good slide images might be acquired manually. The development of specific reactions, with traditional dyes and others, can give important contributions to clarify the chemical nature of the components of nematocytes. The characterisation of nematocyst toxins by staining tests is a goal to achieve.
已描述的钵水母纲物种超过200种,但对其化学和遗传特征进行适当研究的却很少。塔氏霞水母是一种可食用的钵水母,也是欧洲唯一的霞水母科物种,夏季出现在塔霍河和萨多河河口。目前尚未对这两个塔氏霞水母群落进行系统比较,也未对其刺细胞进行化学分析。
为实现这些目标,开发了DNA提取和组织化学染色程序的优化方法。通过核糖体18S、28S和ITS1部分测序,对来自塔霍河和萨多河河口的塔氏霞水母标本进行了比较。利用光学显微镜和电子显微镜研究了刺细胞的形态化学。
宏观和分子结果表明,这两个群落属于同一物种,即塔氏霞水母。苏木精和伊红染色使刺丝囊的发生可视化,并表明在宽刺丝囊的轴以及等刺丝囊和双鞭刺丝囊的小管上的大分子具有碱性特征。通过Masson三色染色法,证实了小管的碱性特性,并提示毒素具有胶原样特征。阿尔辛蓝染色结果表明,刺细胞的外膜可能由含有酸性多糖的大分子组成,这与在水螅中检测到的NOWA和线虫凝集素糖蛋白一致,但也与聚γ-谷氨酸复合物、几丁质样多糖和透明质酸一致。通过冯·科萨试验检测到了钙;其位置表明它与膜的多糖、收缩系统的蛋白质或两者都有相互作用。
DNA提取样品制备的优化可能有助于对鲜为人知的水母物种进行进一步研究。涂片程序的改进简化了浮游动物染色反应的使用。此外,结果表明手动可以获得良好的载玻片图像。利用传统染料和其他染料开发特异性反应,可为阐明刺细胞成分的化学性质做出重要贡献。通过染色试验对刺丝囊毒素进行表征是一个有待实现的目标。