Epperlein H H, Ziegler I, Perris R
Anatomisches Institut der Universität, Freiburg, Federal Republic of Germany.
Cell Tissue Res. 1988 Sep;253(3):493-505. doi: 10.1007/BF00219740.
The purpose of the present investigation was to provide and apply a methodological manual with which the distribution, patterning and relationship of melanophores and xanthophores can be analyzed during early amphibian development. For demonstration of the methods, which include ultrastructural, histochemical and biochemical approaches, Triturus alpestris and Ambystoma mexicanum (axolotl) embryos are used. These two species differ conspicuously in their larval pigment patterns, showing alternating melanophore bands in horizontal (T. alpestris) and vertical (axolotl) arrangements. With transmission- and scanning electron microscopy melanophores and xanthophores were distinguished by their different pigment organelles and surface structures. The presence of phenol oxidase (tyrosinase) was used to reveal externally invisible or faintly visible melanophores by applying an excess of 3,4 dihydroxy-phenylalanine (dopa). Xanthophores were made visible in fixed and living embryos by demonstrating their pterin fluorescence. In addition, pterins were analyzed by HPLC in embryos before and after pigmentation was visible.
本研究的目的是提供并应用一本方法手册，利用该手册可以在两栖动物早期发育过程中分析黑素细胞和黄色素细胞的分布、模式及关系。为演示这些方法，包括超微结构、组织化学和生物化学方法，使用了高山蝾螈和墨西哥钝口螈（美西螈）胚胎。这两个物种的幼体色素模式明显不同，水平排列（高山蝾螈）和垂直排列（美西螈）的黑素细胞带交替出现。通过透射电子显微镜和扫描电子显微镜，根据黑素细胞和黄色素细胞不同的色素细胞器和表面结构对它们进行区分。通过施加过量的3,4 - 二羟基苯丙氨酸（多巴），利用酚氧化酶（酪氨酸酶）的存在来揭示外部不可见或隐约可见的黑素细胞。通过展示黄色素细胞的蝶呤荧光，使其在固定和活体胚胎中可见。此外，在色素沉着可见之前和之后，通过高效液相色谱法对胚胎中的蝶呤进行分析。
