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PubMed1980

The effect of prolonged light and dark adaptation on lens regeneration in the adult newt Triturus viridescens.

Light intensity has been shown to affect lens regeneration in adult newts. Experiments were designed to study the effects of different light controlled environments on lens regeneration in the newt T, viridescens. Animals were preadapted to three light conditions: control, light from below and dark for ninety (90) days prior to lentectomy. The newts remained in the controlled environments post-lentectomy to test the effects of light preadaptation on lens regeneration. The results indicate that controlled light sources from below cannot stimulate the ventral iris to regenerate a new lens or inhibit the dorsal iris from normal lens regeneration. Lens regeneration in preadapted dark animals is similar in rate and amount to the control group indicating a totally dark environment does not always inhibit lens regeneration.

Adaptation, Ocular↗

PubMed1978

Hematologic and hemopoietic alterations following experimental benzene exposure in newts (Triturus cristatus).

Changes in the hematological picture in newts have been investigated utilizing benzene exposure. The first deviation occurs in circulating white cells that become more numerous. Subsequently there is an increase of immature cells on the cortical zone of the liver. About two weeks after the first treatments a decrease of circulating leukocytes begins and the immature white cells continue to increase both in circulation and in the cortical zone of the liver. The red cells present a decrease after one month of treatment. This is preceeded by a decrease in hemoglobin percentage and an increase in the size of red cells and in the number of immature red cells. The data concerning animals treated with an hemolytic agent and with benzene demonstrates an early action of benzene also on red cells. The results suggest a maturation in capacity caused by benzene in both white and in red cells. The intensity of alteration seems proportional to the frequency of treatments. The onset of a particular modification, however, is related to the time interval following the first treatment.

Animals↗

PubMed1976

[Cytochemical analysis of muscle cell histones of Triturus cristatus limbs a) in normal regeneration, b) in regeneration arrested by r-irradiation, and c) in regeneration restored by implants].

The muscular cells of regenerating limbs and of limbs in which regenerative power is restored, show an important decrease in the amount of cytophotometrically detected histones. This decrease is owing to the arginine rich fraction and to the lysine rich fraction. The muscular cells of irradiated limbs show a decrease in the amount of histones. This decrease is owing only to the arginine rich fraction and continues after the thirieth day of irradiation and amputation.

Animals↗

PubMed2006

A molecular assessment of phylogenetic relationships and lineage accumulation rates within the family Salamandridae (Amphibia, Caudata).

We examine phylogenetic relationships among salamanders of the family Salamandridae using approximately 2700 bases of new mtDNA sequence data (the tRNALeu, ND1, tRNAIle, tRNAGln, tRNAMet, ND2, tRNATrp, tRNAAla, tRNAAsn, tRNACys, tRNATyr, and COI genes and the origin for light-strand replication) collected from 96 individuals representing 61 of the 66 recognized salamandrid species and outgroups. Phylogenetic analyses using maximum parsimony and Bayesian analysis are performed on the new data alone and combined with previously reported sequences from other parts of the mitochondrial genome. The basal phylogenetic split is a polytomy of lineages ancestral to (1) the Italian newt Salamandrina terdigitata, (2) a strongly supported clade comprising the "true" salamanders (genera Chioglossa, Mertensiella, Lyciasalamandra, and Salamandra), and (3) a strongly supported clade comprising all newts except S. terdigitata. Strongly supported clades within the true salamanders include monophyly of each genus and grouping Chioglossa and Mertensiella as the sister taxon to a clade comprising Lyciasalamandra and Salamandra. Among newts, genera Echinotriton, Pleurodeles, and Tylototriton form a strongly supported clade whose sister taxon comprises the genera Calotriton, Cynops, Euproctus, Neurergus, Notophthalmus, Pachytriton, Paramesotriton, Taricha, and Triturus. Our results strongly support monophyly of all polytypic newt genera except Paramesotriton and Triturus, which appear paraphyletic, and Calotriton, for which only one of the two species is sampled. Other well-supported clades within newts include (1) Asian genera Cynops, Pachytriton, and Paramesotriton, (2) North American genera Notophthalmus and Taricha, (3) the Triturus vulgaris species group, and (4) the Triturus cristatus species group; some additional groupings appear strong in Bayesian but not parsimony analyses. Rates of lineage accumulation through time are evaluated using this nearly comprehensive sampling of salamandrid species-level lineages. Rate of lineage accumulation appears constant throughout salamandrid evolutionary history with no obvious fluctuations associated with origins of morphological or ecological novelties.

Animals↗

PubMed2005

Phylogeography of two European newt species--discordance between mtDNA and morphology.

The newts Triturus vulgaris and Triturus montandoni are sister species that exhibit contrasting levels of intraspecific morphological variation. Triturus vulgaris has a broad Eurasiatic distribution encompassing both formerly glaciated and unglaciated areas and shows substantial morphological differentiation in the southern part of its range, while T. montandoni, confined to the Carpathians, is morphologically uniform. We analysed sequence variation of two mtDNA fragments of the total length of c. 1850 bp in 285 individuals of both species collected from 103 localities. Phylogenetic analysis of 200 unique haplotypes defined 12 major clades, their age estimated at c. 4.5-1.0 million years (Myr). Most of the older clades were found in the southern part of the range, and also in central Europe, mainly in Romania. The distribution of mtDNA clades points to the existence of several glacial refugia, located in the Caucasus region, Anatolia, the Balkan Peninsula, Italy, and more to the north in central Europe. The concordance between mtDNA based phylogeny and the distribution of T. vulgaris subspecies was weak. Triturus montandoni haplotypes did not form a monophyletic group. Instead they were found in six clades, in five of them mixed with T. vulgaris haplotypes, most likely as a result of past or ongoing hybridization and multiple introgression of mtDNA from T. vulgaris to T. montandoni. Patterns of sequence variation within clades suggested long-term demographic stability in the southern groups, moderate and relatively old demographic growth in the populations inhabiting central Europe, and high growth in some of the groups that colonized northern parts of Europe after the last glacial maximum.

Animals↗

PubMed2004

[Distribution of satellite DNA TkS1 in genomes of salamanders (Salamandridae)].

Genomes of 22 species and subspecies of salamandrids (Triturus, Cynops, Neurergus, Notophthalmus, Pachytriton, and Pleurodeles ) were studied. Satellite TkS1 (32-33 bp) was found in all examined species, both in direct and reversed orientations. The use of single-primer PCR with oligonucleotides, which are homologous to the most conservative part of satellite TkS1, allowed us to reveal DNA fragments, flanked by single copies (or small tandem repeats) of satellites TkS1 having various (5' and 3') orientation. Such fragments were observed in Pleurodeles waltl, Triturus a. alpestris, T. vulgaris lantzi, T. v. vulgaris, Neurergus crocatus and T. vulgaris graecus. The length of amplified DNA fragments in three subspecies of T. vulgaris differed. This might be connected with unequal amounts and different distribution of 5' and 3' copies of satellite TkS1 in their genomes. Patterns of DNA amplification in Triturus a. alpestris and Neurergus crocatus were quite similar. Only two species (Triturus a. alpestris and T. v. vulgaris) had approximately equal amounts and similar distribution of 5' and 3' copies of satellite TkS1 in their genomes.

Animals↗

PubMed1976

Muscle satellite cells in urodele amphibians: faciliatated identification of satellite cells using ruthenium red staining.

The ruthenium red (RR) stained forelimb musculature of three species of urodeles Triturus (Notophthalmus) viridescens, Amblystoma maculatum, Amblystoma opacum in various stages of growth were examined with the electron microscope for the presence of satellite cells. It was found that RR staining facilitated greatly the identification of satellite cells. In young larvae of all three species satellite cells were detected with a frequency of 29% to 48% per total number of nuclei. In adult Triturus and Amblystoma maculatum satellite cells were no longer detected; instead "pericytes" as described by Hay ('74) were seen with a frequency of 12% and 3% respectively. During metamorphosis of Triturus satellite cells, with part of their myofiber-satellite cell intercellular space filled with basement membrane material, occurred at a peak frequency. The cells presumably are intermediate in the formation of "pericytes." At ten days after metamorphosis satellite cells and intermediate cells were no longer detected and the limb musculature contained only "pericytes" similar to the ones observed in adult newts. The significance of the presence of satellite cells in relation to limb regeneration and muscle regeneration is discussed.

Ambystoma↗

PubMed1997

Characterization of pars intermedia connections in amphibians by biocytin tract tracing and immunofluorescence aided by confocal microscopy.

Biocytin, recently introduced in neuroanatomical studies, was used as a retrograde tract tracer in combination with immunofluorescence in order to analyse the neurochemical characters of some central neuronal projections to the pars intermedia in two amphibian species, the anuran Rana esculenta and the urodele Triturus carnifex. After biocytin insertions in the pars intermedia, neurons became retrogradely labelled in the suprachiasmatic hypothalamus and the locus coeruleus of the brainstem in both species. Some scattered biocytin-labelled neurons were observed in the preoptic area. Moreover, working on the same sections, immunofluorescence revealed a number of codistributions and, in some cases, colocalization in the same neurons of biocytin labellings and immunopositivity for (1) tyrosine hydroxylase in the suprachiasmatic hypothalamus and the locus coeruleus of Rana and Triturus, (2) gamma-aminobutyric acid in the suprachiasmatic hypothalamus of Rana and Triturus and (3) neuropeptide Y in the suprachiasmatic hypothalamus of Rana. The specificity of such colocalizations was fully confirmed using dual-channel confocal laser scanning microscopy analysis.

Afferent Pathways↗

PubMed1996

Nitric oxide synthase activity in the olfactory bulb of anuran and urodele amphibians.

Nitric oxide synthase activity was studied by means of NADPH-diaphorase activity and nitric oxide synthase immunoreactivity in the main and accessory olfactory bulbs of the frog Rana perezi and the newt Triturus marmoratus. In both species, NADPH-diaphorase staining was observed in all olfactory fibers. Vomeronasal fibers were NADPH-diaphorase-positive in Triturus but they were NADPH-diaphorase-negative in Rana. Nitric oxide synthase immunoreactivity was not observed in the primary afferents in any case. Granule cells were NADPH-diaphorase-positive and nitric oxide synthase-immunopositive in the main and accessory olfactory bulb of Rana, and in the main olfactory bulb of Triturus. The homogeneous NADPH-diaphorase staining of olfactory fibers is similar to what has been reported in teleosts, and it contrasts with the spatial segregation of NADPH-diaphorase-positive and -negative olfactory projections in rodents. These results confirm the interspecies variability of the NADPH-diaphorase/nitric oxide synthase distribution in the olfactory system of vertebrates.

Animals↗

PubMed1974

Analysis of the C-value paradox by molecular hybridization.

Poly(A)-containing RNA was isolated from ovaries of Xenopus laevis laevis and Triturus cristatus carnifex and used as a template for the synthesis of radioactive complementary DNA with RNA-dependent DNA polymerase. When annealed with an excess of homologous DNA, the complementary DNA is rendered double-stranded with kinetics that suggest that the coding sequences are single-copy in both these organisms. In Triturus, these sequences are distinct from the majority of the genome, which consists of repeated sequences, and distinct from the ribosomal cistrons, which are present in proportion to the increase in C-value relative to the Xenopus genome. Moreover, the number of different poly(A)-containing molecules in the ovary (sequence complexity) is the same in Xenopus and in Triturus.

Adenine Nucleotides↗

PubMed1992

High resolution scanning electron microscopy of the nuclear envelope: demonstration of a new, regular, fibrous lattice attached to the baskets of the nucleoplasmic face of the nuclear pores.

The nuclear envelope (NE) of amphibian oocytes can be readily isolated in relatively structurally intact and pure form and has been used extensively for structural studies. Using high resolution scanning electron microscopy (HRSEM), both surfaces of the NE can be visualized in detail. Here, we demonstrate the use of HRSEM to obtain high resolution information of NE structure, confirming previous data and providing some new information. NEs, manually isolated from Triturus cristatus oocytes, have been mounted on conductive silicon chips, fixed, critical point dried and coated with a thin, continuous film of chromium or tantalum and viewed at relatively high accelerating voltage in a field emission scanning electron microscope with the sample within the objective lens. Both nucleoplasmic and cytoplasmic surfaces of the nuclear pore complexes (NPC) have been visualized, revealing the cytoplasmic coaxial ring, associated particles, central plug/transporter and spokes. The nucleoplasmic face is dominated by the previously described basketlike structure attached to the nucleoplasmic coaxial ring. In Triturus, a novel, highly regular flat sheet of fibers, termed the NE lattice (NEL) has been observed attached to the distal ring of the NPC basket. The NEL appears to be distinct from the nuclear lamina. Evidence for the NEL is also presented in thin TEM sections from Triturus oocytes and GVs and in spread NEs from Xenopus. A model is presented for NEL structure and its interaction with the NPCs is discussed.

Amphibians↗

PubMedDate not supplied

Expression of PCNA positivity in the brain of normal adult heterothermic vertebrates: further observations.

As part of our study of non-experimentally induced encephalic proliferation in unequivocally adult individuals of several heterothermic Vertebrates (Podarcis sicula, Triturus carnifex, Rana esculenta, Carassius carassius), we deal here with areas not considered in previous investigations, i.e. various encephalic regions (except the telencephalon) in Podarcis sicula, Triturus carnifex and Rana esculenta, the diencephalon and medulla oblongata in Carassius carassius, and the olfactory bulbs in the two Amphibians. In the previous and current research, we have used Proliferating Cell Nuclear Antigen (PCNA) as a marker. PCNA is a ubiquitous intracellular antigen of the cycline family (proteins that regulate the cell cycle), which acts as an auxiliary protein to DNA polymerase delta; it can be detected immunocytochemically with monoclonal antibodies to reveal cell cycle phases that coincide with DNA synthesis. Spontaneous proliferation events, revealed by PCNA positivity, were constantly present in this study, being substantial in the olfactory region and diencephalon, very modest in the mesencephalon and myelencephalon, and absent in the cerebellum. In particular, signs of proliferation were abundant in the epithelium lining the cavities of the olfactory bulbs, while they were of different magnitude in tracts (with multiple and comparatively different sites related to the dorsal and/or ventral thalami) of the ependyma that delimits portions of the III ventricle and also, in all the species examined, at the level of the preoptic and infundibular recesses. Such signs were rare in the ependymal epithelium of the mesencephalic ventricle in Podarcis sicula and the rhombencephalic ventricle in all four species examined. This immunoreactivity was also observed in extra-ependymal areas: in the internal granular layer of the olfactory bulbs in Triturus carnifex and Rana esculenta; in the diencephalic nuclei of the habenula in Podarcis sicula, in both Amphibians and in Carassius carassius; in the mesencephalic tectum in Podarcis sicula and in the two Amphibians. As in our previous studies, the current immunocytochemical picture revealed by PCNA positivity generally agrees with literature reports on the presence of normal proliferation in the areas investigated here. These literature sources consist primarily of the observations of Kirsche (1967), emerging from his preceding experimental investigations, and of confirmatory data from studies in subsequent decades by other researchers obtained with tests different from our marker. Nevertheless, the number of studies that deal with the species considered in the present research, or species closely related to them, is rather limited.

Aging↗

PubMed1978

A portion of all major classes of histone messenger RNA in amphibian oocytes is polyadenylated.

Ovaries of the amphibian Triturus viridescens contain a considerable amount of 7 to 12 S RNA which fractionates with poly(A)+ RNA on oligo(dT)-cellulose column chromatography and directs the synthesis of all five classes of histones in a wheat germ cell-free system. The polyadenylate tracts associated with this 7 to 12 S poly (A)+ RNA are heterogeneous in length, ranging from approximately 60 to 120 nucleotides. Partially purified subfractions of histone mRNA templates were isolated from this RNA by preparative polyacrylamide gel electrophoresis. The correlation between the poly(A) content, the template activity in vitro, and the cell-free products encoded by these RNA subfractions suggests that Triturus ovary RNA contains poly(A)+ histone mRNA. The 7 to 12 S poly(A)+ RNA, but not 7 to 12 S poly(A)- RNA, is an effective template for the avian myeloblastosis virus reverse transcriptase-directed synthesis of complementary DNA (cDNA) in the presence of oligo(dT) primer. In the absence of primer, virtually no cDNA is synthesized. When cDNA complementary to 7 to 12 S poly(A)+ RNA is hybridized in situ to Drosophila melanogaster polytene chromosome preparations. cDNA hybrids are found primarily in the region 39D-E, the locus of the histone genes in Drosophila. This cDNA also hybridizes to the sea urchin histone gene sequences contained in the chimeric bacterial plasmids pSp2 and pSp17. These results provide strong evidence for the reality of polyadenylated histone mRNA in Triturus ovary.

Animals↗

PubMed1995

The role of vertical and planar signals during the early steps of neural induction.

The classical Einsteck-test (Spemann and Mangold, Roux Arch. Dev. Biol. 100: 599-638, 1924) and data from total exogastrulae (Holtfreter, 1933) suggest that vertical signals are transmitted between the chordamesoderm (organizer) and reacting ectoderm in the early phase of neural induction. In contrast to these results with Axoloti (urodeles), several authors observed the expression of neural specific genes in Xenopus exogastrulae, isolated dorsal blastopore lip with adjacent ectoderm (open-face explants) and Keller-sandwiches. Our data with Xenopus (anurans) also show that the expression of neural specific genes takes place in exogastrulae. However, when we prepared open face explants and exogastrula-like structures by microdissection at very early gastrula stage, the signal of a class II beta-tubulin, characteristic of terminal neural differentiation, is not found in the ectoderm. These results suggest that planar signals transmitted from the chordamesoderm into the ectodermal part can fairly be excluded under these experimental conditions. In similar experiments with Triturus alpestris we could not observe either the differentiation of neural structures in the ectodermal part of exogastrulae. These results confirm earlier experiments of Holtfreter performed with Ambystoma mexicanum (Axoltl) embryos. On the basis of the published data of different authors and our results, we cannot exclude the existence of planar signals for early and/or transient expressed genes before the onset of gastrulation in Xenopus, which make the neuroectoderm susceptible for the response to vertical signals during gastrulation. On the other hand our experiments with Triturus alpestris suggest that planar neural signals are unlikely in this species. These differences between Triturus and Xenopus embryos are discussed in the context of the peculiarities in morphological structure, competence and speed of development of the two species.

Ambystoma mexicanum↗

PubMed1996

Factors responsible for the establishment of the body plan in the amphibian embryo.

A central topic of embryology is the establishment of the body plan during embryogenesis. Starting with maternal factors distributed in the early cleavage stages in distinct patterns and gradients cell-to-cell interactions including early embryonic induction result in the formation of mesoderm and the organizer area. While many facts are known about the role of growth factors like activin (closely related to the vegetalizing factor), processed Vg1, BMPs and FGF for mesoderm formation, the establishment of the central nervous system is not yet well understood. However, there is growing evidence that neural induction is a multistep process at the level of the dorsal mesoderm (organizer) and the reacting neuroectoderm. Therefore the existence of only one neuralizing factor is unlikely. We report about data that follistatin protein is not a direct neural inducer. Furthermore our comparative studies of Xenopus and Triturus exogastrulae indicate that planar signals are unlikely in the Triturus embryo (urodeles) during the early steps of neural induction. Vertical signals emanating from the chordamesoderm are essential for the terminal neuralization and regionalization of the central nervous system during gastrulation for both Xenopus and Triturus. The putative role of neuralizing factors and BMP/activin-like molecules for the stabilization or shift of neuroectoderm into different pathways of differentiation (epidermis or neural default state) is discussed.

Amphibians↗

PubMed1983

Water balance of five amphibian species at different stages and phases, as affected by hypophysial hormones.

Five amphibian species were studied for the effect of hypophysial hormones on their water balance. The species were three anurans, Rana ridibunda, Bufo viridis and Pelobates syriacus, and two urodeles Salamandra salamandra and Triturus vittatus. In the first four species different stages of development were studied, in the newt both the terrestrial and aquatic phases of the adult were examined. The hormones used were oxytocin (OXY), arginine vasotocin (AVT) and prolactin (PL). Oxytocin caused most water retention when compared with the other hormones, especially responding were juveniles of Rana and Bufo, but also the terrestrial phase of the adult newt Triturus. Arginine vasotocin affected mostly juvenile Pelobates. Prolactin caused water retention in juvenile Rana and in the terrestrial phase of Triturus. In general the hormones affected the juvenile stages more than either larvae or adults.

Amphibians↗

PubMed1975

Microfilaments in the external surface layer of the early amphibian embryo.

A comparison was made by transmission electron microscopy of the microfilaments in the surface layers of the early embryos of Triturus alpestris and Xenopus laevis at stages of development up to neurulation. Actin-like filaments which bound heavy meromyosin (HMM) were found in cell extracts of all stages, but were comparatively rare in the newly fertilized egg. Ten nm microfilaments were present throughout development in Xenopus, and from the mid-neurula stage in Triturus. Both kinds of microfilament were located in the circumferences of superficial ectoderm cells at the level of the apical junctions, the 10 nm microfilaments in association with desmosomes which began to develop before gastrulation in Xenopus. The accumulations of microfilaments in the apical constrictions, which form in ectoderm cells of Triturus early gastrulae when dissociated in a calcium-free medium, suggest that they are contractile elements. In the absence of such accumulations in the cell apices, the reverse curling exhibited by Xenopus ectodermal explants is attributed rather to a separation of the cells' lateral borders. Cytochalasin B (5 mug/ml) caused ectodermal explants from the early gastrulae of both species to disaggregate. With the rupture of the apical junctions there was a disorganization of the associated microfilamentous layer.

Actins↗

PubMed1978

The effects of alpha and beta adrenergic agents on spleen cell antigen binding in four amphibian species.

Adults of two urodele amphibian species (Triturus cristatus carnifex and Cynops hongkongensis) and two anuran species (Rana temporaria and Xenopus laevis laevis) were immunized with a 25% suspension of sheep or horse erythrocytes. After eight or 14 days, splenic lymphocytes were removed, and their specific red cell-binding capacities tested by immunocytoadherence. Antigen-binding cells were classified as high-dose nonsecretory (S-) or secretory (S+), according to whether they bound a single layer or several layers or erythrocytes. The stimulation of both alpha and beta adrenoreceptors reduced the numbers of S+ rosettes formed by Triturus and Cynops lymphocytes, whereas a beta agonist increased and an alpha agonist decreased S+ rosette formation by Rana and Xenopus splenic lymphocytes. These effects were blocked by alpha and beta adrenoreceptor antagonists. Low-dose immunization of Xenopus with a 0.0025% suspension of sheep erythrocytes gave a minimal number of S+ rosettes two and eight days after immunization, and beta adrenoreceptor stimulation had no effect on antigen binding. These results are discussed in terms of the distribution of alpha and beta adrenoreceptors in amphibians and possible relationships between S+ and high-dose S- antigen-binding cells, and support the view that functional lymphocyte heterogeneity exists in these lower vertebrates.

Amphibians↗