The Sexual Behaviour of the Great Crested Newt,Triturus cristatus (Amphibia: Salamandridae)

The sexual behaviour of Triturus cristatus is described. The first stage of sexual behaviour consists of an orientation phase in which the male approaches the female, sniffs her and moves in front of and perpendicular to her. The second stage is a prolonged period of static display with two different tail movements, called Fan and Lash, in 13.8 ± 1.3 bouts (x? ± SE) with intervals of 10.5 ± 0.3 s between bouts. Each bout contains 6.7 ± 0.3 regular Fan beats at a frequency of 0.5–0.8 Hz. This Fanning provides olfactory and mechanical stimulation to the female. 15% of bouts also contain a violent Lash, in which the male rapidly slaps the female's flank. Some Lashes are used to stop the female when she tries to move away. In the third stage of the sexual sequence, the male turns away, Creeps for 6.7 ± 0.3 s (x? ± SE) and then Quivers his tail. A receptive female follows and touches his tail, and the male deposits a spermatophore and then turns to one side with his tail folded along his flank in a position called Brake. The female now approaches again and the male steps away sideways in a movement called Rebrake. After several Rebrakes the male stops, the female touches his tail and the spermatophore may now be picked up in her cloaca. This sequence can be repeated several times during one encounter. This behaviour is discussed in relation to previous studies of courtship in this genus.     

Heterochromatin characterization of sex chromosomes in Triturus marmoratus (Urodela, Salamandridae).

The sex chromosomes of the Iberian marbled newt, Triturus marmoratus, were studied using various banding techniques, including restriction enzyme/nick translation (RE/NT) procedures. Four types of heterochromatin on the sex chromosomes could be distinguished: (1) distamycin A/DAPI and chromomycin A3/distamycin A positive, EcoRI/NT negative, and HaeIII/NT and HinfI/NT positive; (2) distamycin A/DAPI and chromomycin A3/distamycin A positive, but RE/NT negative; (3) AT rich, but RE/NT negative; and (4) distamycin A/DAPI and chromomycin A3/distamycin A positive, EcoRI/NT and HinfI/NT negative, but HaeIII/NT positive. These data suggest a common origin for the terminal heterochromatic domains of both the X and Y chromosomes in this species.     

Banding patterns on lampbrush chromosomes of Triturus marmoratus (Amphibia Urodela) by the Giemsa stain

Lampbrush chromosome preparations from the newt species Triturus marmoratus have been submitted to a banding procedure by using a Giemsa stain technique (C-banding) as well as variants of the method. Centromeres, most of telomeres, the nucleolus organizing region and some segments along the chromosome axes appear to be differently stained. The centromere positions have been indicated on the maps of the lampbrush complement of the species. The possible relationships between banding and chromosome structure and organization are briefly discussed.     

Field observations of the sexual behaviour of the smooth newt, Triturus vulgaris vulgaris (Amphibia: Salamandridae)

This study reports observations of smooth newts, Triturus vulgaris vulgaris , breeding in a pond in southern England. Efforts were made to collect data on the timing of oviposition and on sexual behaviour during the breeding seasons of 1985 and 1986. Despite some between-year differences in the timing of certain events, the qualitative pattern of reproduction was similar in both years. Courtship interactions were seen most frequently during a relatively short period of time before females began laying their eggs, which occurred in a highly synchronized manner. It seems likely that the probability of any one courtship encounter resulting in spermatophore transfer was low. Males competed for mates by chasing females and by interfering with one another's courtships, but no overt aggression was seen. Females appeared to find such male-male interactions 'aversive'. Scramble competition between males was most intense when the majority of females in the population were laying their eggs and were unresponsive to courtship. The mating system of the smooth newt most closely resembles a lek in which the intensity of sexual selection among males varies as a function of female availability.     

The Cloaca and Spermatheca of the Female Smooth Newt, Triturus vulgaris L. (Amphibia: Urodela: Salamandridae)

The anatomy and histology of the cloaca and spermatheca of 13 female smooth newts ( Triturus vulgaris ) were studied by light microscopy. The cloaca consists of a short, anterior tube (which receives the oviducts), which opens into a larger, more posterior chamber. The spermatheca, which is the only gland in the cloaca, consists of a mass of exocrine, acinar tubules which empty individually and dorsolaterally into the posterior part of the cloacal tube and all but the most posterior part of the cloacal chamber. Stored sperm are most abundant in the spermathecal tubules during the peak of the breeding season (in May and June); during their period of storage, the sperm do not appear to make intimate contact with the epithelial cells lining the spermathecal tubules. Longterm storage of viable sperm from one breeding season to the next is probably absent in the smooth newt. Sperm storage by and multiple insemination of females both occur in this species and are necessary preconditions for competition between the sperm of different males for the fertilization of eggs. However, sperm competition has not been demonstrated in the smooth newt.     

High aquatic niche overlap in the newts Triturus cristatus and T. marmoratus (Amphibia,Urodela)

We studied spatial niche metrics of large-bodied newts (Triturus cristatus and T. marmoratus) in three breeding ponds in western France. Adults and larvae were sampled with underwater funnel traps. Larvae were identified to the species with diagnostic microsatellite DNA markers. The distribution of adult T. cristatus and T. marmoratus across pond regions differed in one out of six cases, no differences were observed between larvae (two ponds studied). Niche overlap and niche breadth indices across resource states defined as pond regions or individual traps were high (Schoener's C: pond regions 0.60–0.98, traps 0.35–0.71; Levins' B: pond regions 0.71–0.98, traps 0.35–0.76). Adults of large-bodied newts significantly differed in resource use from small-bodied newts (T. helveticus). The results are discussed in view of the occurrence of interspecific breeding attempts, and the unpredictable ecological characteristics of newt breeding ponds.     

Experimental hybridization within the genus Triturus (Urodela: Salamandridae)

Spermatogenesis in the F₁ hybrid (2n=24=12 +12 ) between the closely related newt species T. cristatus carnifex and T. marmoratus was apparently normal up to pachytene. Many unpaired chromosomes were present at diplotene and a typical diakinesis was lacking. Primary spermatocytes at meta-and meta-anaphase contained up to 12 regular intergenomal bivalents and a corresponding number of univalents when less then 12 II. Most chiasmata were terminal or subterminal, some intercalary. Chiasmata between corresponding heterospecific chromosomes can be reported as true: real crossing over has taken place, proving the presence of primary chromosomal homologies between the 2 sets of the parental species. Evidence for recombination is based on the segregation of particular markers (i.e., subterminal C-bands and NORs) observed in certain chromosomes at metaphase II. One chromatid of single chromosomes can show the T. cristatus phenotype and the other the T. marmoratus phenotype. A few primary spermatocytes contain a certain number of irregular associations (intragenomal or intrahaploid bivalents, irregular intergenomal bivalents, chromosome multivalents) joined by chiasmata which can be defined as anomalous. Other abnormalities concern the occurrence of interlocked bivalents which occasionally show an anomalous exchange between heterologous chromatids. — Cytogenetic criteria useful to evaluate the taxonomic relationships between different species have been discussed as well as some possible trends in chromosome evolution and speciation within the genus Triturus.     

Breeding dynamics of Triturus carnifex at a pond in northwestern Italy (Amphibia, Urodela, Salamandridae)

A population of the Italian crested newt ( Triturus carnifex ) was studied in 1984 at a pond near Druento (Turin, Piedmont, northwestern Italy). Animals entering and leaving the pond fell into paired pit-fall traps during both spring and autumn migrations. A positive correlation found between rainfall and migration, a shortening of migration periods and summer inactivity suggest that, in the Mediterranean region, air moisture may be a limiting factor for newt activity. During their aquatic period adults showed strong individual differences in weight changes that cannot be explained on the basis of initial body weight or length. High variability in crested newt life history traits may be the result of repeated bottlenecks caused by environmental stress and high population isolation.     

The cycle of follicular and interstitial cells (Leydig cells) in the testis of the marbled newt, Triturus marmoratus (Caudata, Salamandridae)

Ultrastructural examination of the marbled newt (Triturus marmoratus) testis throughout the annual cycle revealed that during the period of testicular quiescence (November-February), primordial germ cells proliferate within cords of filament-rich epithelial cells that will become follicular cells (FCs). Fibroblast-like cells surround the FCs and form the lobule-boundary interstitial cells (ICs). During the period of germ cell development from primordial germ cells to round spermatids (March-June), the FCs surrounding the developing germ cells contain scanty cytoplasm with abundant rough endoplasmic reticulum and scarce filaments. With spermatid elongation (July-August), the FC size grows, its nucleus becomes irregularly outlined, and its cytoplasm displays abundant smooth endoplasmic reticulum, residual bodies, lipid droplets, and large vacuoles. After spermatozoon release by the FCs (August-September), the adjacent ICs increase their size and transform into Leydig cells with abundant smooth endoplasmic reticulum, mitochondria with tubular cristae, and lipid droplets. During the period of testicular quiescence (November-February), the Leydig cells undergo involution, eventually developing the morphological attributes of mesenchymal cells. Intermingled among these cells, cords of filament-rich cells are observed. During this period of the cycle, spermatozoon cysts supported by FCs are present. At the beginning of the germ cell proliferation period (March), these spermatozoa are released, and the adjacent ICs undergo a transformation into Leydig cells similar to those observed in August-September. Maturation and involution of ICs occur when testosterone levels are known to be rising and falling, respectively.     

Celestial orientation in the marbled newt (Triturus marmoratus)

Orientation toward breeding ponds plays an important role in the seasonal movements of amphibians. In this study, adult marbled newts were tested in a circular arena to determine sensory cues used to locate breeding ponds. Animals were collected from a temporary pond situated in northern Spain, taken to the experimental site 340 m distant, and tested for orientation under a variety of conditions (i.e., orientation under a clear night sky, orientation under an overcast night sky, and orientation under a clear night sky in the presence of an altered geomagnetic field). These investigations have demonstrated that the marbled newt is able to orient using celestial cues. Animals chose a compass course in the direction of their breeding pond only when celestial cues were available. Conversely, the ambient geomagnetic field does not seem to be relevant to orientation of marbled newts since they were unable to orient themselves using the ambient geomagnetic field in the absence of celestial cues. Electronic Publication     

Intermediate heterochromatic zones in Triturus (Amphibia: Caudata)

Employment of the C-banding procedure disclosed the presence of slightly stained bands in some chromosome regions of several Triturus species. These regions have been previously described in other organisms as 'grey C-bands'. Possible explanations for the presence of these bands in Triturus karyotypes are discussed.     

Two dispersed highly repeated DNA families of Triturus vulgaris meridionalis (Amphibia,Urodela) are widely conserved among Salamandridae

Two BamHI families of repeated sequences were characterized from the genome of the Italian smooth newt, Triturus vulgaris meridionalis (Amphibia, Urodela). The first family, which is divided into subfamilies, consists of tandemly arranged arrays whose basic repeat is around 398 bp long; these arrays are dispersed throughout the entire chromosome sets of the various species of Triturus tested. Moreover the family is widely conserved among Salamandridae, being detected by genomic DNA blotting of Notophthalmus viridescens, Taricha granulosa, Salamandrina terdigitata and Euproctus platycephalus. The second BamHI family is represented by a cloned sequence of 419 bp, which is dispersed in the chromosome set of several species of Triturus. The sequence is also conserved in S. terdigitata and in E. platycephalus but is not detectable in N. viridescens or T. granulosa. The cloned sequence is most probably only part of a longer unit interspersed within the Triturus genome.by H.C. Macgregor     

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.     

First Evidence of an Altitudinal Variation in the Brain Size of Triturus alpestris alpestris (Laur.) (Amphibia,Caudata, Salamandridae)

This study deals with geographical variations in brain-body weight relationships in the alpine newt, Triturus alpestris alpestris. According to the canonical axis, C.A., the mean allometry in this species is 0.473±0.03; an allometry value of 0.5 is considered to be characteristic of Caudata. No significant difference was observed between the four reduced major axes (R.M.A.) of lowland animal samples (mean value of the pooled samples, C.A. = 0.469±0.210). In contrast, between the mountain samples a significant difference exists, the R.M.A. in the sample from l'Alpe d'Huez (Isère, France) being 0.331±0.57 as compared to 0.72±0.18 in the sample from Durmitor (Montenegro, Yugoslavia). In spite of this difference, their encephalisation index (Ie) did not differ significantly, having on average a value of 118.18±8.5, against īe = 93.8±8.0 for lowland samples. A significant difference in encephalisation index between mountain and lowland alpine newts thus exists. This is probably the first observation in vertebrates of an encephalisation index increase with altitude. A biological hypothesis is proposed to explain this peculiar observation.     

Heterochromatic DNA in Triturus (Amphibia,Urodela)

The MspI family of highly repeated sequences is a centromeric satellite DNA representing about 1% of the genome of the Italian smooth newt, Triturus vulgaris meridionalis. We have studied the structure, genomic organization, chromosomal localization and conservation across species of this family. MspI sequences are around 197 bp long, as shown by sequencing of three cloned units. The family is organized in large clusters of tandemly arrayed units, present at almost all the centromeres of T.v. meridionalis, and is well conserved in the T.v. vulgaris subspecies. Conserved MspI sequences are also present in the related species T. helveticus, where they appear to be clustered at the centromeres of only a few chromosomes. MspI sequences are not found in other Triturus species analysed. The correlation of these sequences with the overall distribution pattern of heterochromatin and the extent of their conservation within the genus Triturus, are discussed.     

Chromosome location of the genes for 28S, 18S and 5S ribosomal RNA in Triturus marmoratus (Amphibia Urodela)

The localization of the 28S, 18S and 5S rRNA genes in the mitotic chromosomes, and of the 5S rRNA genes in the lampbrush chromosomes of Triturus marmoratus has been studied by RNA/DNA in situ hybridization. The 28S and 18S genes are located in a subterminal position, and the 5S genes in an intermediate position, on the long arm of mitotic chromosome X. In situ hybridization on lampbrush chromosomes has shown that the 5S genes are located at or near a dense matrix loop landmark. The cytogenetic implications of these findings are briefly discussed.     

The maps of the lampbrush chromosomes of Triturus (Amphibia urodela)

The lampbrush chromosomes of Triturus vulgaris meridionalis were isolated from the germinal vesicle of medium and large-sized oocytes and studied with phase-contrast microscope. The maps were constructed on the basis of the lengths and major morphological features of the chromosomes. The length of each map is equal to the mean of the relative lengths of the corresponding chromosome from different oocytes (the relative length of each chromosome is represented by the ratio between its absolute length and that of chromosome XII from the same complement, conventionally considered as 100 units long). The maps arranged in decreasing length order, were oriented according to the most frequent position of chiasmata, as centromeres were not always evident. — Chromosomes VI and XI bear a sphere in subterminal position. Landmarks typical for T. vulgaris meridionalis are the loops inserted on chromosomes VIII (47 units), X (23 units), XI (34 units) and XII (34 units) frequently presenting themselves under the form of double loop bridges of considerable extension. On chromosomes I (4 units), VI (13 units), X (4 units) and XI (36 units) giant bodies were found that are sometimes comparable to dense-matrix loops. Chromosome XI includes a nucleolus-organizing region, sometimes identifiable by the presence of an inserted nucleolus. Normal and granular loops (much extended at times), axial granules, globules, and loopless bars supplement the morphology of the lampbrush chromosomes of this species.     

Spermathecae of Salamandrina terdigitata (Amphibia: Salamandridae): Patterns of sperm storage and degradation

The spermathecae of female Salamandrina terdigitata were observed using light and transmission electron microscopy during the fallspring period of sperm storage and secretory activity and during the summer stasis. When sperm are stored inside the spermathecae, the product synthesized by spermathecal epithelial cells is exported into the lumen, where it bathes the sperm. During sperm storage some spermatozoa undergo degradation by the spermathecal epithelium. This process, which includes sperm capture by the apical microvilli, formation of endocytic vacuoles and production of lysosomes, becomes prominent shortly after oviposition. In many instances, cells filled with vacuolized spermatozoa and/or residual bodies undergo desquamation from the spermathecal epithelium and enter the lumen together with residual sperm. Desquamated cells, together with residual sperm, are a common feature in the spermathecal lumina at the end of the egg-laying season. Concomitant to the activity of the spermathecal epithelium, macrophages move into the spermathecae from the stroma and contribute to the degradation of both the residual sperm and desquamated epithelial cells. As a result of this degradation activity, spermathecae observed during the short summer stasis appear devoid of secretory product and sperm. By late summer, however, the spermathecae already show early signs of an imminent resumption of biosynthetic activity. © 1995 Wiley-Liss, Inc.