Weber J
Zoological Institute, University of Zurich-Irchel, Switzerland.
J Cell Sci. 1995 Jan;108 ( Pt 1):403-12. doi: 10.1242/jcs.108.1.403.
The rhodamine derivatives tetramethyl-rhodamine-5/6-maleimide (TROMI) and tetramethyl-rhodamine-6-isothiocyanate (TRITC) were allowed to react with living Hydra vulgaris. The two fluorescent dyes stain the polyps to different degrees, apparently without impairing their viability and behaviour. Concerning nematocytes, TROMI preferentially couples to cytoskeletal elements only of mounted nematocytes whereas TRITC selectively reacts with structural components of cysts of late nematoblasts, which thereafter develop apparently normally into mature nematocytes. Hence TROMI-labelling indicates that nematocytes are mounted and ready for discharge; TRITC-labelling can be used as a tool to investigate the final maturation, migration and installation of nematocytes in Hydra. Together with a new non-fixative method to dissociate Hydra polyps into single, identifiable cells, the two labelling methods allow direct quantitative dynamic studies of nematocyte turnover and open new possibilities of investigating the regulation and the mechanisms of nematocyte supply and migration.
使罗丹明衍生物四甲基罗丹明-5/6-马来酰亚胺(TROMI)和四甲基罗丹明-6-异硫氰酸酯(TRITC)与活体普通水螅发生反应。这两种荧光染料对水螅虫体的染色程度不同,显然并未损害它们的活力和行为。关于刺细胞,TROMI仅优先与已附着的刺细胞的细胞骨架成分结合,而TRITC则选择性地与晚期刺丝囊母细胞的囊泡结构成分发生反应,此后这些细胞显然能正常发育成成熟的刺细胞。因此,TROMI标记表明刺细胞已附着并准备好释放;TRITC标记可作为一种工具,用于研究水螅中刺细胞的最终成熟、迁移和定位。这两种标记方法与一种将水螅虫体解离成单个可识别细胞的新型非固定方法相结合,能够对刺细胞更新进行直接定量动态研究,并为研究刺细胞供应和迁移的调控及机制开辟了新的可能性。