有毒的丝带蠕虫,Cephalothrixcf.simula(Palaeonemertea,Nemertea),具有极高浓度的河豚毒素(TTX)。尽管在该物种的卵中发现了TTX,尚未探索动物个体发育中毒素的命运。这里,使用高效液相色谱串联质谱和免疫组织化学与抗TTX抗体,我们检查了水平,profile,和TTX及其类似物(TTXs)在C.cf幼虫中的定位。整个受精后41天的模拟。用光学和电子显微镜对TTX积累部位的细胞进行了详细研究。新孵化的幼虫在所有细胞中都具有弱的TTX样免疫反应性。随着后续的发展,外胚层结构中TTX标记的强度,中胚层细胞和顶腺的顶柱增加。在外胚层结构中,在多纤毛中观察到强烈的TTX标记,II型颗粒状,I型粘液,和表皮的基底细胞,在口腔腺体的III型颗粒细胞中。在中胚层,TTX位于肌肉和单粒薄壁组织样细胞中。C.的卵和幼虫。simula包含五个TTX,有两种主要毒素-TTX和5,6,11-三脱氧TTX。TTXs的水平和相对比例在发育阶段之间没有显着差异,确认幼虫从母体卵中获得毒素并能够保留它。这项研究的结果为C.cf的TTX轴承装置的形成提供了见解。模拟物通过幼虫发育。
The toxic ribbon worm, Cephalothrix cf. simula (Palaeonemertea, Nemertea), possesses extremely high concentrations of tetrodotoxin (TTX). Although
TTX has been found in the eggs of this species, the fate of the toxin in the ontogeny of the animal has not been explored. Here, using high performance liquid chromatography with tandem mass spectrometry and immunohistochemistry with anti-
TTX antibodies, we examined levels, profile, and localization of
TTX and its analogues (TTXs) in larvae of C. cf. simula throughout 41 days post-fertilization. A detailed investigation of cells in sites of TTX-accumulation was performed with light and electron microscopy. Newly hatched larvae possessed weak
TTX-like immunoreactivity in all cells. With subsequent development, intensity of TTX-labeling in the ectodermal structures, mesodermal cells and apical cylinder of the apical gland increased. In the ectodermal structures, an intense TTX-labeling was observed in the multiciliated, type II granular, type I mucoid, and basal cells of the epidermis, and in the type III granular cells of the mouth gland. In the mesoderm,
TTX was localized in the muscle and unigranular parenchyma-like cells. Eggs and larvae of C. cf. simula contained five TTXs, with two major toxins - TTX and 5,6,11-trideoxyTTX. Level and relative proportion of TTXs did not differ significantly among developmental stages, confirming that larvae obtained toxins from maternal eggs and were able to retain it. The results of this study provide insights into the formation of TTX-bearing apparatus of C. cf. simula through the larval development.