Identification of pigment cells during early amphibian development (Triturus alpestris,Ambystoma mexicanum) |
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Authors: | Hans-Henning Epperlein Irmgard Ziegler Roberto Perris |
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Institution: | (1) Anatomisches Institut der Universität, Freiburg, Germany;(2) Gesellschaft für Strahlen- und Umweltforschung, Institut für Experimentelle Hämatologie, München, Germany;(3) Developmental Biology Center, University of California, Irvine, California, USA;(4) Anatomisches Institut, Albertstr. 17, D-7800 Freiburg, Germany |
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Abstract: | Summary 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.Abbreviations DOPA
dihydroxy-phenylalanine
- FCS
fetal calf serum
- FIF
formaldehyde-induced fluorescence
- FITC
fluorescein isothiocyanate
- HPLC
high performance liquid chromatography
Dedicated to the memory of Dr. Michael Claviez |
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Keywords: | Neural crest Melanophores Xanthophores Pigments Triturus alpestris Ambystoma mexicanum (Amphibia Urodela) |
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