Identification of pigment cells during early amphibian development (Triturus alpestris,Ambystoma mexicanum)

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     

A new type of cell in the pulmonary epithelium of the newt Triturus alpestris Laur

Summary Single cells of a new type appear scattered among pneumocytes in the pulmonary epithelium. The surfaces of these cells communicate with the air space and display numerous finger-like microvilli. In comparison to pneumocytes, these cells have a more lucid cytoplasm and their apical parts contain large amounts of electron-lucent vesicles and electron-dense granules, which are probably released into the lumen of the lung. These secretory cells exhibit a yellow formaldehyde-induced fluorescence, which suggests that they belong to the class of APUD cells.     

Chromosome differentiation in the Triturus alpestris complex (Amphibia: Caudata)

Several individuals from distinct populations of 7 subspecies included in the Triturus alpestris complex have been analysed to obtain characteristic C-banding patterns.The differences found among the populations as regards the amount, complexity and distribution of heterochromatin bands have been employed to test a pattern of chromosome evolution.Relevant similarities and dissimilarities of these populations are discussed in the light of the tentative evolutionary history inferred from a rooted tree.     

The Ectomesenchymal-Endodermal Interaction-System (EEIS) of Triturus alpestris in Tissue Culture

A piece of head neural fold tissue from behind the prospective ear region of a Triturus alpestris neurula was cultivated, together with a piece of ventrolateral pharynx endoderm from the same neurula, in hanging drop cultures. This system, referred to as the Ectomesenchymal-Endodermal Interaction-System (EEIS), offers insight into the visible processes of attachment, migration and differentiation of neural crest cells emigrating from the piece of neural fold.
The neural crest cells were not found to move preferably in the direction of the pharynx endoderm. Therefore, instead of chemotaxis, the concept of contact inhibition¹ provides a more satisfactory explanation for the distribution pattern of emigrating neural crest cells.
During culture, the neural crest cells differentiate into neuroblasts, pigment cells, myoblasts, chondroblasts and, after about 11 days, into perichondrial cells. After 6 days, a large number of neural crest cells, now called mesenchymal cells, persist without any visible differentiation throughout the culture.
Chondroblasts only develop from neural crest cells which have been in contact with the pharynx endoderm, as opposed to all other crest cells differentiating in the EEIS.     

Vitamin A promotes DNA synthesis and mitosis in blastemas of Triturus alpestris

Vitamin A palmitate orally administered to young, post-metamorphic Triturus alpestris (Amphibia-Urodela), for 4 and 7 days after bilateral forelimb amputation at the middle of the zeugopodium, enhanced the mean DNA content of the blastema cells. Moreover, the number of cells in mitosis was nearly twice as high in vitamin A treated animals than in control ones.     

Autoradiographic evaluation of vitamin A effects on Triturus alpestris blastema cells

Autoradiographic studies on Triturus alpestris regenerates treated with vitamin A palmitate for 4, 7, 11 and 14 days after amputation, reveal a considerable diminution in the percentage of cells duplicating DNA, as judged by measurements of labelling indices. Based on previous observations, that vitamin A promotes DNA-synthesis, mitosis and growth of regenerates, we conclude that the lower labelling indices calculated are due to an acceleration of S-phase processes, resulting in a concomitant reduction in its duration.     

Beeinflussung des Eingeweidesitus beim Alpenmolch (Triturus alpestris) durch elektrischen Strom

Zusammenfassung Neurula-Keime (Harrison-Stadien 15–17) vom Alpenmolch wurden in einem Elektrophoresegerät 1 Std lang der Wirkung eines elektrischen Stromes ausgesetzt. Unter den im Larvenstadium fixierten Versuchstieren fanden sich Eingeweide-Irreguläre und -Inverse in einem gegenüber Kontrollen stark erhöhten, statistisch gesicherten Prozentsatz.

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Influence of the electrical current upon the situs viscerum of triturus alpestris

Summary Neurulae (Harrison 15–17) ofTriturus alpestris were kept within an electrical set up for 1 hour. At a later stage the larvae showed irregularities and inversions of their viscera in a much higher frequency than the untreated controls.

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Durchgeführt im Rahmen des Forschungsvertrages 041-65-10 BIO D mit Euratom. Mit Unterstützung der Deutschen Forschungsgemeinschaft durch Leihapparate im Heiligenberg-Institut.     

The Ectomesenchymal-endodermal Interaction System (EEIS) of Triturus alpestris in Tissue Culture

Differentiation of visceral cartilage was studied in the Ectomesenchymal-endodermal Interaction System (EEIS), a hanging-drop culture system containing a piece of head neural fold tissue from behind the prospective ear region of a Triturus alpestris neurula together with a piece of ventrolateral pharynx endoderm from the same embryo. Cartilage cells differentiated only from those neural crest cells which have been in contact with the pharynx endoderm. Cartilage differentiation occurred neither in cultures containing neural fold or pharynx endoderm alone nor in distance cultures in which both explants were separated from each other at a greater distance than usual. However, in some critical cultures of the EEIS containing pharynx endoderm, excised more ventrally from the regular site, accumulations of ectomesenchymal cells were observed near the endoderm which were not transformed into cartilage.
The differentiation of cartilage was classified into three stages: (1) prechondroblasts (day 1 to 5), (2) chondroblasts (about day 6) and (3) chondrocytes (about day 10).
The data obtained suggested that cellular contact between the two inductively correlated tissues was a prerequisite for the transmission mechanism while mitotic cell divisions were mainly responsible for the spread of the inductive message within the responding ectomesenchymal cells.     

Endocrine-like cells in the lungs of the newt,Triturus alpestris Laur.

Summary Endocrine-like cells (ELC) scattered in the basal part of the ciliated epithelium were identified in the lungs of the newt, Triturus alpestris. These ELC have a clear cytoplasm containing large amounts of secretory vesicles (69–180 nm in diameter), especially in their basal parts, but do not display formaldehyde-induced fluorescence. The ELC may be associated with nerve fibres.     

Relatedness,body size and paternity in the alpine newt,Triturus alpestris

Sexual selection has traditionally been investigated assuming that male quality is as skewed as patterns of male reproductive success can sometimes be. Recently, female choice has been investigated under the model of genetic compatibility, which assumes that each individual female has her own 'best' mate and there is no overall optimal choice for all females. We investigated female mate choice in the newt species Triturus alpestris, a member of a genus where female choice has been investigated only within the context of the optimal male (female choice for condition-dependent traits). We provided females with two males that differed in one condition-dependent trait (body size) and overall genetic composition. Both male body size and female body size did not influence paternity, but the degree of genetic relatedness between females and potential mates did. Two components of fitness (fecundity and hatching success) did not differ between singly and multiply sired clutches, indicating that females do not employ polyandry as a means of increasing offspring fitness through genetic bet-hedging. Instead, we hypothesize that females may mate initially for fertility assurance, but prefer less-related males as the most genetically compatible mates.     

Influence of vitamin A palmitate on the growth of regenerating Triturus alpestris forelimbs

The objective of this investigation was to analyse the effects of vitamin A palmitate orally administered to young postmetamorphic Triturus alpestris on the growth of regenerates. Vitamin A was administered for 4, 7, 11 and 14 days after amputation at the middle of the right zeugopod. Growth of regenerates was evaluated on the basis of external size measurements and blastema cell proliferation. Calculation of the percentages of cells in each cell-cycle phase and estimation of mitotic indices during the period of drug administration were performed by microdensitometric scanning of Feulgen-stained nuclei and counting of chromosome configurations, respectively. Vitamin A was found to induce polyploidism, to enhance transition from G1- to S-phase cell nuceli, to prolong the duration of G2-phase to the detriment of S-phase and to promote mitotic activity in the mesenchyme cells. In accordance with the above, an acceleration in the growth rates of treated blastemas with respect to controls was observed.     

Skull of the neotenic salamandrid amphibian Triturus alpestris and abbreviated development in the Tertiary Salamandridae

The skull of neotenic individuals of the Alpine Newt Triturus alpestris from the locality Drakolimni (Greece) is described on the basis of models made from magnified serial frontal sections. In order to recognize features associated with neoteny and paedomorphosis, the results were compared with normal development of contemporary Triturus and of other Caudata. The neotenic larvae from Greece correspond to advanced stages of metamorphosis in normal development of Triturus alpestris. Comparison with salamandrids from the Tertiary of Europe in which the hyobranchial skeleton was preserved, namely Brachycormus noachicus, Chelotriton paradoxus, and Palaeopleurodeles hauffi, revealed that both latter taxa were completely metamorphosed adults anatomically similar to their contemporary relatives Tylototriton, Echinotriton, and Pleurodeles, whereas Brachycormus, though apparently related to Chelotriton, was a neotenic amphibian. This is suggested by its incompletely metamorphosed but ossified hyobranchial skeleton. © 1996 Wiley-Liss, Inc.