Kolberg K J, Martin V J
Department of Biological Sciences, University of Notre Dame, Indiana 46556.
Development. 1988 Jun;103(2):249-58. doi: 10.1242/dev.103.2.249.
Planula larvae of Halocordyle disticha were examined for the presence of catecholamines using a multipronged approach. Transmission electron micrographs of planular sensory cells and ganglionic cells demonstrated dense-cored vesicles and electron-dense droplets in both cell types. These vesicles and droplets were similar in morphology to catecholamine-containing granules of vertebrates. Planulae processed with the SPG histofluorescence technique, specific only for catecholamines, exhibited blue-green fluorophores which were most prominent in the anterior ectoderm. Such fluorescence was associated with sensory cells, ganglionic cells and the neural plexus. Pretreatment of planulae with neuropharmacological agents which prevent reuptake (reserpine) or cause release (nicotine, ephedrine) of catecholamines caused a diminution of the fluorophores. Pretreatment of animals with 6-hydroxydopamine, which causes destruction of catecholamine-containing cells, prevented any fluorescent response. Ultrastructural examination of reserpine-treated planulae revealed a dramatic reduction in the populations of dense-cored vesicles and electron-dense droplets. Furthermore, many of the vesicles and droplets remaining in reserpinized animals appeared washed out, i.e. stained faintly. Exposure of planulae to exogenous norepinephrine caused premature, rapid metamorphosis and produced polyps with slightly stunted tentacles and pitted, irregular hypostomes. Exposure of planulae to nicotine caused similar effects. Rearing planulae in sea water containing alpha blockers, phentolamine and tolazoline, had no discernible effect on behaviour (motility, phototactic response) or gross morphology. However, planulae raised in sea water containing propranolol, a beta blocker, ceased all movement, became tack-shaped and died within 72 h. These results meet multiple criteria for the identification of catecholamines in hydrozoan planulae and suggest that such catecholamines may function as neurotransmitters, neurohormones or neuromodulators during larval development.
采用多管齐下的方法,对双叉扁螅的浮浪幼虫进行了儿茶酚胺检测。浮浪幼虫感觉细胞和神经节细胞的透射电子显微镜图像显示,这两种细胞类型中均存在有致密核心的囊泡和电子致密液滴。这些囊泡和液滴在形态上与脊椎动物含儿茶酚胺的颗粒相似。用仅对儿茶酚胺有特异性的SPG组织荧光技术处理的浮浪幼虫,呈现出蓝绿色荧光团,在前外胚层最为明显。这种荧光与感觉细胞、神经节细胞和神经丛有关。用可阻止儿茶酚胺再摄取(利血平)或导致其释放(尼古丁、麻黄碱)的神经药理学药物预处理浮浪幼虫,会导致荧光团减少。用6-羟基多巴胺预处理动物,可导致含儿茶酚胺细胞的破坏,从而阻止任何荧光反应。对利血平处理的浮浪幼虫进行超微结构检查发现,有致密核心的囊泡和电子致密液滴的数量显著减少。此外,利血平处理动物中剩余的许多囊泡和液滴似乎被洗脱,即染色很浅。将浮浪幼虫暴露于外源性去甲肾上腺素会导致过早、快速变态,并产生触手略发育不良、口盘有凹痕且不规则的息肉。将浮浪幼虫暴露于尼古丁也会产生类似的效果。在含有α受体阻滞剂酚妥拉明和妥拉唑啉的海水中饲养浮浪幼虫,对其行为(运动性、趋光反应)或总体形态没有明显影响。然而,在含有β受体阻滞剂普萘洛尔的海水中饲养的浮浪幼虫停止了所有运动,变成了大头针形状,并在72小时内死亡。这些结果符合鉴定水螅纲浮浪幼虫中儿茶酚胺的多个标准,并表明此类儿茶酚胺在幼虫发育过程中可能作为神经递质、神经激素或神经调节剂发挥作用。