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The influence of tail autotomy on agonistic behaviour in a territorial salamander.

Assessment of potentially asymmetrical characters (such as fighting ability and resident advantage) is often important in determining the outcome of agonistic interactions. Loss of body parts, a predator defence mechanism used by many animals, may lead to a reduction in fighting ability and may be easily assessed by competitors. We investigated the influence of tail loss on the expression of agonistic behaviour in the territorial red-backed salamander, Plethodon cinereus. Residents and intruders were matched for body size, and pairs were tested in all combinations of tailed or tailless residents with tailed or tailless intruders. Neither residents nor intruders altered their behaviour based on their own tail condition, but they did alter their behaviour based on the tail condition of their opponents. Intruders showed more aggression or less submission towards tailless residents than towards tailed residents. When contests were between residents and intruders of the same tail condition (both tailed or tailless), intruders were more aggressive towards residents when both were tailless than when both were tailed, indicating that tail loss does not directly hamper aggressive displays. In contests where the asymmetry between residents and intruders was small (based on tail condition and residency status), intruders showed more aggression and less submission than in contests where the asymmetry was large. Residents did not differ in their behaviour for most comparisons. Thus, for intruders, the tail condition of residents is an important determinant of agonistic behaviour displayed in territorial contests. For residents, factors other than tail condition (such as resource value) may be more likely to influence their behaviour. Copyright 1998 The Association for the Study of Animal Behaviour. Copyright 1998 The Association for the Study of Animal Behaviour.

Journal Article↗

Pigment patterns of larval salamanders (Ambystomatidae, Salamandridae): the role of the lateral line sensory system and the evolution of pattern-forming mechanisms.

In many species of salamanders, pigment cells derived from the neural crest give rise to a horizontal stripe pattern in hatchling larvae. A defining element of these horizontal stripe patterns is a region over the middle of the myotomes that is relatively free of melanophores. This study shows that formation of a "melanophore-free region" and horizontal stripe pattern in Ambystoma tigrinum tigrinum (family Ambystomatidae) correlates with the development of the trunk lateral line sensory system. Moreover, prevention of lateral line development results in greater densities of melanophores in the middle of the flank, essentially eliminating the melanophore-free region in this taxon. A phylogenetic survey also revealed that ablation of the lateral lines has qualitatively similar effects on melanophores in seven of eight additional taxa (Ambystomatidae: A. barbouri, A. maculatum, A. talpoideum; Salamandridae: Notophthalmus viridescens, Pleurodeles waltl, Taricha granulosa, T. rivularis). In Taricha torosa, however, a superficially similar melanophore-free region forms prior to lateral line development, and ablation of the lateral lines does not perturb the horizontal stripe pattern. Finally, heterospecific grafting experiments demonstrated that T. torosa lateral lines are competent to generate a melanophore-free region, and T. torosa melanophores are competent to respond to cues associated with the lateral lines. These results indicate that lateral line-dependent pattern-forming mechanisms are common and probably ancestral within the families Ambystomatidae and Salamandridae and suggest that these ancestral mechanisms have been retained in T. torosa as redundant, lateral line-dependent mechanisms for stripe formation have evolved.

Ambystomatidae↗

When neural crest and placodes collide: interactions between melanophores and the lateral lines that generate stripes in the salamander Ambystoma tigrinum tigrinum (Ambystomatidae).

A prominent element of the early larval pigment pattern in the salamander Ambystoma tigrinum tigrinum (family Ambystomatidae) is a horizontal stripe over the lateral surface of the myotomes where otherwise abundant, neural crest-derived melanophores are not found. This study examines the formation of this "melanophore-free region". When the trunk lateral lines were ablated (by removing cranial lateral line placodes), the melanophore-free region did not form; instead, melanophores populated the middle of the flank and the distribution of yellow, neural chest-derived zanthophores was perturbed. Time-lapse videomicrography demonstrated that during normal development, the melanophore-free region is established because melanophores retreat from the midbody lateral line primordium as it migrates caudally along the inner side of the epidermis. Melanophores do not repopulate the middle of the flank after primordium migration and heterochronic grafting experiments suggest that extracellular factors contribute to maintaining the melanophore-free region during these later stages. Finally, photographic series, microsurgical manipulations, electron microscopy, and staining for molecules of the extracellular matrix (peanut agglutinin-binding components, tenascin, chondroitin sulfate proteoglycans, fibronectin, laminin) suggest that several factors contribute to establishing and maintaining the melanophore-free region, including steric effects of the lateral lines, interactions between melanophores and xanthophores, lateral line-dependent alterations of the subepidermal basement membrane, and a general elaboration of the extracellular matrix. Lateral line effects on melanophores are inferred to be a shared, ancestral feature of pigment pattern development for the families Ambystomatidae and Salamandridae (D.M. Parichy, Dev. Biol. 174, 265-282. 1996). The results of this study thus provide insights into a phylogenetically primitive mechanism for stripe formation, and a context for interpreting evolutionary innovations in pattern-forming mechanisms.

Ambystoma↗

Contribution of ventral and dorsal mesoderm to primitive and definitive erythropoiesis in the salamander Hynobius retardatus.

Previously, we found that the conversion of hemoglobins (Hbs) from the larval to the adult type occurred within a single erythroid cell population in a salamander, Hynobius retardatus ("Hb switching" model), whereas the transition involves replacement of red-blood-cell (RBC) populations ("RBC replacement" model) in many amphibians (M. Yamaguchi, H. Takahashi, and M. Wakahara, 2000, Dev. Gene Evol. 210, 180-189). To further characterize the Hb transition, developmental changes in the erythropoietic sites have been intensively analyzed using larval- and adult-specific globin antibodies and globin and GATA-3 RNA probes. Cells of the ventral blood island (VBI) and the dorsolateral plate (DLP) in embryos differentiate in situ to erythroid cells that contain larval globin mRNA, suggesting that both the VBI and the DLP contribute to "primitive" erythropoiesis. In contrast, the expression pattern of the GATA-3 gene suggests that cells of the DLP may contribute to "definitive" hematopoiesis. In order to determine whether it is possible to define a definitive erythropoiesis in H. retardatus or not, further experiments were done: (1) when metamorphosing larvae were treated with phenylhydrazine to induce anemia and then bled at the postmetamorphic stage after recovery from the anemia, a precocious Hb transition was observed in these animals; (2) an RBC population expressing only adult Hb was confirmed by subtracting the number of RBCs expressing larval Hb from the total number of RBCs during metamorphosis. All these results support the existence of a definitive erythroid cell population that contributes only adult RBCs in this species.

Amino Acid Sequence↗

Seasonal changes in plasma concentrations of sex steroids in the salamander Hynobius nigrescens.

Changes in plasma concentrations of androgens, estradiol-17 beta, and progesterone were examined throughout the year in Hynobius nigrescens. The highest mean concentration of androgens (312.7 ng/ml) was found in males in the process of spermiation and having just entered the breeding pond in early spring. Plasma estradiol-17 beta concentration was at its base level in females both before and after the animals had entered the pond. Plasma progesterone concentrations were low throughout the year except during the aquatic phase in March when females possessed some ova within the coelom and many ova in the oviducts and ovisacs after the completion of ovulation. During the terrestrial phase, or nonbreeding season, plasma concentrations of androgens and estradiol-17 beta were in approximate correspondence with the development of sex accessory structures. This is the first documentation of plasma concentrations of sex steroids in hynobiid salamanders.

Animals↗

Interactions of androgens and estradiol on sex accessory ducts of larval tiger salamanders, Ambystoma tigrinum.

Immature tiger salamander larvae were treated with 12.5 or 25 micrograms of estradiol, testosterone, or dihydrotestosterone (DHT), or 12.5 micrograms of estradiol combined with 12.5 micrograms of either testosterone or DHT. Müllerian duct epithelium was more stimulated by combined steroid treatment than by any steroid alone. Estradiol antagonized the action of DHT in the Wolffian duct. Both of the androgens and estradiol when administered alone at the higher dose stimulated enlargement of connective tissue surrounding the ducts, but the combined 12.5 micrograms androgen/12.5 micrograms estrogen treatment was more effective even though the total steroid administered was the same. The effectiveness of DHT on müllerian cells of this species is evidence against a required aromatization of androgens to explain paradoxical steroid effects and suggests that fundamental differences may exist in steroid receptors of müllerian ducts, connective tissue, and Wolffian ducts. A possible role for the urodele duct system for assessing estrogenic activity of environmental contaminants is discussed.

Ambystoma↗

Interactions of gonadal steroids and pesticides (DDT, DDE) on gonaduct growth in larval tiger salamanders, Ambystoma tigrinum.

In view of the current worldwide decline in amphibian populations, exploratory studies are needed to assess the potential for environmental contaminants to act as endocrine disrupters of the amphibian reproductive system. The present study investigated the effects of DDT dichlorodiphenyltrichloroethane (DDT) and dichlorodiphenyldichloroethylene (DDE) on the development of amphibian gonaducts. Larval male and female tiger salamanders (Ambystoma tigrinum), with immature gonads, were immersed in a sublethal solution of p,p'-DDE or technical-grade DDT (80% p,p'-DDT and 20% o,p'-DDT). Additionally, larvae were injected with the steroid hormones estradiol or dihydrotestosterone (DHT). Morphometrics were used to analyze the effects and interactions of steroid and pesticide treatments on larval gonaducts. Estradiol and DHT stimulated cell proliferation and hypertrophy of the müllerian duct epithelium in both sexes. Wolffian duct epithelium, however, was stimulated only by DHT treatment. The pesticide DDT antagonized the estrogenic actions of the steroid treatments, and p,p'-DDE acted as an estrogen on the müllerian ducts of females only. The müllerian ducts of males, and the wolffian ducts of both sexes, were unaffected by DDT or DDE alone. While confirming the previously reported estrogenic actions of estradiol and DHT on urodelean gonaducts, the results contradict the expected estrogenic actions of DDT and antiandrogenic actions of p,p'-DDE. Instead, in A. tigrinum, technical-grade DDT had an antiestrogenic action and p,p'-DDE an estrogenic action.

Animals↗

Humoral regulation of hemoglobin transition from larval to adult types in a salamander, Hynobius retardatus.

In amphibians hemoglobin (Hb) transition from larval to adult types occurs during metamorphosis. Hemoglobin transition in the salamander Hynobius retardatus also occurs during metamorphosis, but depends on activity of the pituitary gland, rather than that of the thyroid gland. These findings were supported by the fact that the transplanted pituitary gland to hypophysectomized (Hx) larvae exerted accelerating effects on Hb transition. Contrary to this, a homogenate of the pituitary gland from Xenopus laevis had no accelerating effect on Hb transition when injected into Hx larvae of H. retardatus. Whereas exogeneously applied triiodothyronine (T3) or a combination of T3 and hydrocortisone had an accelerating effect on Hb transition in the Hx larvae, a single treatment with hydrocortisone in the Hx larvae had no effect. When an inhibitor of synthesis of adrenocortico-steroid hormones, metyrapone, was applied in combination with goitrogens to intact larvae, Hb transition from larval to adult types was suppressed compared with normal controls, suggesting that the thyroid-corticoid system is involved in Hb transition. These results suggest that Hb transition in H. retardatus is controlled by two independent accelerating factors, from both the pituitary gland and the thyroid-corticoid system. These control mechanisms are different from the general regulation of metamorphosis that occurs in many amphibians. A tentative model of hormonal controls in Hb transition from larval to adult types during the metamorphosis in H. retardatus is presented.

Adrenal Cortex Hormones↗

Ancestry of an isolated subspecies of salamander, Ambystoma tigrinum stebbinsi Lowe: the evolutionary significance of hybridization.

Most phylogenetic systematists assume speciation results in dichotomously branching phylogenies. Hybridization that gives rise to a new lineage can produce character homoplasty that might obscure a species' true history. We report the results of a restriction-enzyme analysis of mitochondrial DNA (mtDNA) variation in three tiger salamander subspecies (Ambystoma trigrinum mavortium, Ambystoma tigrinum nebulosum, and Ambystoma trigrinum stebbinsi) and compare the results to studies of morphological and allozymic variation in these taxa. Allozymically, A. t. mavortium and A. t. nebulosum share most of their genomes (although each has several unique alleles), yet color pattern and mtDNA haplotypes are distinct. Color pattern and allozyme data suggest that A. t. stebbinsi shares a common ancestor with A. t. mavortium, while the A. t. stebbinsi mtDNA haplotype is derived from an A. t. nebulosum haplotype. Thus, our data suggest that A. t. stebbinsi originated through hybridization between A. t. mavortium and A. t. nebulosum. That hybridization can produce recognizably distinct evolutionary entities has long been recognized for plants, but the evolutionary significance of hybridization in animals should be examined more closely. Conservation agencies must recognize that hybrids and hybrid taxa are not necessarily evolutionary "mistakes," and they might have significant importance in the production of natural biodiversity.

Animals↗

Molecular phylogenetic relationships of neotropical salamanders of the genus Pseudoeurycea.

Pseudoeurycea, with 34 described species, is one of the most diversified groups of neotropical salamanders. I generated a phylogenetic hypothesis of relationships for Pseudoeurycea and related taxa, based on DNA sequences of 16S, Cyt b, and ND4 mitochondrial genes. The analyses include 27 species of Pseudoeurycea and samples from the monotypic Lineatriton, Ixalotriton, and Parvimolge. All phylogenetic analyses resulted in a paraphyletic Pseudoeurycea. Ixalotriton and Pseudoeurycea parva always form a monophyletic group. P. parva is transferred to Ixalotriton based on morphological and molecular grounds. The phylogenetic position of the newly defined Ixalotriton clade is uncertain since it is part of an unresolved basal polytomy. Parvimolge is closely related to Pseudoeurycea, and it is also part of the basal polytomy. Lineatriton, a highly specialized taxon, is deeply nested within Pseudoeurycea. In order to provide a taxonomic arrangement consistent with the monophyly of the different units that reflects both evolutionary history and morphological specialization, Pseudoeurycea should be split into several taxonomic units.

Animals↗

A tandemly repeated DNA family originated from SINE-related elements in the European plethodontid salamanders (Amphibia, Urodela).

We have characterized a highly repetitive family, named Hy/Pol III, in the genome of the European salamanders Hydromantes (Plethodontidae). This family consists of short, tandemly repeated sequences organized in clusters, scattered through the genome as shown both by in situ hybridization to chromosomes and by Southern blot hybridization. The repeat unit is about 200 bp in length and it is a composite element since it contains a SINE-like retroposon with a tRNA structure, flanked by two short direct repeats. The whole element itself is bordered by two other direct repeats. The sequence data suggest that two elements, presumably derived from polymerase III transcripts, have been inserted one into the other, giving rise to the observed composite structure. During evolution the Hy/Pol III family was then amplified by tandem duplication at the DNA level. The inferred relationships between Hy/Pol III members from three representative species of the European Hydromantes suggests that a subfamily structure characterizes the evolutionary history of this family.

Animals↗

Optic nerves in plethodontid salamanders (amphibia, urodela): neuroglia, fiber spectrum and myelination.

In five species of lungless salamanders, family Plethodontidae, which all show highly developed visual abilities, the ultrastructure of the optic nerve was investigated and the total number of retinal ganglion cell axons, the percentage of myelinated axons, and the volume densities of glia and axons were determined. More than 80% of all axons were smaller than 0.4 micron and only 2-3% were larger than 0.8 micron. In individual nerves the degree of myelination varied between 1 and 9% which is in the range reported for other amphibian species. The miniaturized and highly paedomorphic species Batrachoseps attenuatus was an exception because only very few or even no myelinated axons were present in the nerve, which is unique among gnathostome vertebrates. The five investigated species had total numbers of axons ranging from 26,000 in Batrachoseps attenuatus to about 50,000 in Plethodon jordani. These numbers are the lowest found among vertebrates with an elaborated visual system. The amount of glial material in the optic nerve varied between 25 and 50%, with larger nerves possessing more glia than smaller ones. Ultrastructural analysis revealed that the optic nerve of each species contained both astrocytes and oligodendrocytes, although often in immature form. In Batrachoseps attenuatus the glia showed features of both astrocytes and oligodendrocytes which reflect an undifferentiated state.

Animals↗

The lateral spread of signal between bipolar cells of the tiger salamander retina.

When mapped with a small spot of light, the central receptive fields of bipolar cells in the salamander retina are much larger than the extent of bipolar cell dendrites. Furthermore responses of bipolar cells to distant spots of light are considerably delayed relative to proximal spots. Using quantitative modelling, electrical coupling between bipolar cells is examined and rejected as a sufficient explanation of the data. An active process appears to shape signal waveform as signals spread laterally in the bipolar cell layer. Chemical synaptic coupling between bipolar cells is considered and shown to be inconsistent with the data. It is suggested that local, transient negative feedback from amacrine cells is involved in shaping bipolar cell signals.

Animals↗

Observations on the blood-testis barrier in a frog and a salamander.

A blood-testis barrier has been demonstrated in a frog, Rana esculenta, and in a salamander, Salamandra salamandra, using lanthanum as an electron-dense marker during fixation. The tracer penetrates the interstices between somatic follicle or Sertoli cells and germ cells in regions of the testis containing spermatogonia and spermatocytes, up to the level of punctate tight junctions. The latter can be localized between the somatic cells that line seminiferous units containing spermatids and mature spermatozoa. The barrier thus appears to be established after meiosis in both species investigated, although spermatids of different developmental stages can be found in open compartments of the testis in S. salamandra.

Animals↗

Regional differences in cell density and cell genesis in the olfactory epithelium of the salamander, Ambystoma tigrinum.

The olfactory epithelium undergoes continuous regeneration. The present quantitative study uses tritiated thymidine autoradiography to investigate regional differences in the rate of olfactory epithelial cell genesis in the tiger salamander. There was a significant gradient in the incorporation of thymidine from the posterior to the anterior in the nasal cavity: the posterior epithelium underwent cell genesis much faster than the anterior. Additionally, the posterior epithelium was thinner and contained fewer cells than the anterior, although the proportions of receptor, supporting and basal cells remained about the same throughout the epithelium. After 5 or 20 days most of the labelled cells were found in the basal cell layer, although there were a few labelled supporting cells. This confirms observations in other species that there are two populations of dividing cells in the olfactory epithelium: the basal cells which give rise to receptor cells, and the supporting cells. The gradients in epithelial thickness, receptor cells, and the rate of cell genesis parallel a gradient in responsiveness to odorants observed in electrophysiological studies (Mackay-Sim et al. 1982; Mackay-Sim and Shaman 1984). The significance of these anatomical and physiological gradients is presently unclear.

Ambystoma↗

The light microscopic localization of substance P- and somatostatin-like immunoreactive cells in the larval tiger salamander retina.

Light microscopic immunocytochemistry was utilized to localize the populations of substance P (SP)- and somatostatin (SOM)-like immunoreactive cells in the larval tiger salamander retina. Of 104 SP-immunostained cells observed, 82% were Type 1 amacrine cells. Another 8% of the SP-cells were classified as Type 2 amacrine cells, while 10% of the SP-cells had their cell bodies located in the ganglion cell layer and were designated as displaced amacrine cells. Each type of SP-like immunoreactive cell was observed in the central and peripheral retina. SP-immunopositive processes were observed in the inner plexiform layer as a sparse plexus in sublamina 1 and as a denser network of fibers in sublamina 5. Seventy-eight percent of the 110 somatostatin-immunopositive cells observed were designated as Type 1 amacrine cells. Another 12% of SOM-cells were classified as displaced amacrine cells, while only two SOM-immunopositive Type 2 amacrine cells were observed. Nine percent of the SOM-cells were designated as interplexiform cells, based on their giving rise to processes distributing in the outer plexiform layer as well as processes ramifying in the inner plexiform layer. Each type of SOM-immunoreactive cell was observed in the central and peripheral retina, with the exception of the Type 2 amacrine cells, whose somas were only found in the central retina. Lastly, SOM-immunopositive processes in the inner plexiform layer appeared as a fine plexus in sublamina 1 and as a somewhat denser network of fibers in sublamina 5.

Ambystoma↗

Scanning electron microscopy of microcorrosion casts of the vascular bed in the skin of the spotted salamander, Salamandra salamandra L.

Microcorrosion casts of blood vessels in the skin of the spotted salamander, Salamandra salamandra L., were studied using scanning eletron microscopy. The investigated vessels include a subepidermal network of respiratory capillaries and the vessels of poison glands. A hypothesis is proposed, according to which both types of vascular beds possess a common origin. Probable factors involved in the differentiation of the primary netword of the subepidermal vessels in larvae are indicated and speculation concerning the supposed mosaic type of circulation in the capillaries of the subepidermal respiratory bed is presented.

Animals↗

The blocking effect of l-cis-diltiazem on the light-sensitive current of isolated rods of the tiger salamander.

The effect of the organic compound l-cis-diltiazem on the light-sensitive current of isolated rods of the tiger salamander was analysed by rapidly changing the extracellular medium using the method of Hodgkin et al. (1985). Addition to the extracellular medium of small amounts of l-cis-diltiazem rapidly inhibits the photocurrent. Complete suppression of the current was observed with 1 mM l-cis-diltiazem. Half blockage of the photocurrent occurred with about 150 microM l-cis-diltiazem. The blocking effect of l-cis-diltiazem was enhanced by light and by a reduction of extracellular Na. A concentration of l-cis-diltiazem of 140 microM, which suppresses one third of the photocurrent, was able to completely suppress the photocurrent carried by Ba. It is suggested that l-cis-diltiazem blocks the light-sensitive channel, possibly competing with cyclic guanosine-3'-5'-monophosphate (cGMP) for an internal regulatory site.

Ambystoma↗