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PubMed1999

In vitro nitric oxide effects on basal and gonadotropin-releasing hormone-induced gonadotropin secretion by pituitary gland of male crested newt (Triturus carnifex) during the annual reproductive cycle.

The objective of this study was to test the possible nitric oxide (NO) involvement in pituitary gonadotropin secretion in the male crested newt, Triturus carnifex. Pituitaries were incubated in vitro with medium alone, GnRH, NO donor (NOd, sodium nitroprusside), NO synthase inhibitor (NOSi, Nomega-nitro-L-arginine methyl ester), cGMP analogue (cGMPa, 8-bromo-cGMP), soluble guanylate cyclase inhibitor (sGCi, cystamine), GnRH plus NOSi, GnRH plus sGCi, and NOd plus sGCi during the annual reproductive cycle: pre-reproduction, reproduction (noncourtship and courtship), and the refractory, recovery, and estivation periods. To determine pituitary gonadotropin secretion indirectly, newt testes were superfused in vitro with preincubated pituitaries, and androgen release was determined. NO synthase (NOS) activity and cGMP levels were assessed in the preincubated pituitaries. Medium alone- and GnRH-preincubated pituitary increased androgen secretion during pre-reproduction, noncourtship, courtship, and recovery; the GnRH-induced increase was higher than the medium alone-induced increase during pre-reproduction, noncourtship, and recovery. NOd and cGMPa increased androgens in all reproductive phases considered except courtship; the NOd- and cGMP-induced increase was higher than the medium alone-induced increase during pre-reproduction, noncourtship, and recovery. NOS activity was highest during courtship and lowest during the refractory and estivation periods. GnRH increased NOS activity during pre-reproduction, noncourtship, and recovery. Cyclic GMP levels were highest during courtship and lowest during the refractory period and estivation. GnRH increased cGMP levels during pre-reproduction, noncourtship, and recovery, while NOd did so during all reproductive phases considered. These results suggest that basal and GnRH-induced gonadotropin secretion are up-regulated by NO in the pituitary gland of the male Triturus carnifex.

Androgens↗
PubMed1977

Early steroid metabolism in Xenopus laevis, Rana temporaria and Triturus vulgaris embryos.

Embryos of Xenopus laevis, Rana temporaria and Triturus vulgaris exposed to radioactive pregnenolone have been found to convert it to progesterone. Incubations with radioactive progesterone showed that it was actively metabolized by oocytes and embryos. In Xenopus incubations progesterone was converted to 5alpha-pregnane-3,20-dione, 17alpha-hydroxy-4-pregnen-3-one, 4-androstene-3,17-dione and 17alpha-20alpha-dihydroxy-4-pregne-3-one, indicating 5alpha-reductase, 17alpha-hydroxylase, 19-20-desmolase and 20alpha-hydroxylase activities. In oocytes of Triturus and Rana no evidence of 19-20-desmolase was found. In Rana oocytes were also not evidence of 17alpha-hydroxylase activity. All identified activities except 20alpha-hydroxylase were common to embryos of all three species. It is suggested that the steroid enzyme activities present in the embryos are not solely derived from the oocytes but synthentized during early development. Possible meaning of this kind of metabolism during differentiation remains open.

Animals↗
PubMed1988

The first sequenced normal hemoglobin lacking histidine in position 146 of the beta-chains. The primary structures of the major and minor hemoglobin components of the great crested newt (Triturus cristatus, Urodela, Amphibia).

The hemoglobin of the Great Crested Newt (Triturus cristatus), an animal maintaining the gas exchange to about 85% through the skin, consists of a major (HbM = 65%) and a minor (Hbm = 35%) component. The primary structures of the four chains are presented. They could be separated by reversed-phase HPLC and were cleaved with trypsin and additionally by acid hydrolysis. Both the native chains and their peptides were sequenced by liquid and gas phase sequenators. At the N-terminus the alpha M-chains are by one amino-acid residue longer and the beta M-chains by one residue shorter, resulting in a chain length of 142 and 145, respectively. The alpha m-chains are of normal length whereas in the beta m-chains the C-terminal histidine in position 146 is missing. Both alpha-chains differ by 50 residues (35.2%) and the beta-chains by 63 (43.2%). The alpha-chains were compared with those of other salamandroid hemoglobins. The difference to human hemoglobin is marked by 61 (43.3%) amino-acid substitutions in both alpha-chains and by 78 (53.4%) in both beta-chains. Numerous heme contacts and positions involved in the subunit interface are affected by replacements. The most interesting of them were studied by molecular modeling. The importance of the missing beta m-146(HC3)His and of the substitution of several amino-acid residues involved in the binding of organic phosphates is discussed with respect to the reduced Bohr effect of Triturus cristatus hemoglobin.

Amino Acid Sequence↗
PubMed2000

Evolutionary relationships among Europan newts (genus Triturus) as inferred from two mtDNA fragments.

European newts (genus Triturus) are widely studied, but their phylogeny is not yet unambiguously resolved. Fragments of mitochondrial DNA experiencing different rates of evolution (the ATPase and 12S rDNA genes) were sequenced in order to test a phylogenetic hypothesis derived from biochemical and behavioural data. Well supported branches of the existing phylogeny also gained support in our study. Within the subgenus Palaeotriton (the group of small-bodied newts) the monophyletic origin of the hypothesized T. boscai-T. italicus clade remained ambiguous, whereas strong support was gained for the sister-taxon relationship of T. vulgaris and T. montandoni. The position of T. vittatus within the subgenus Triturus as a sister taxon to the clade of big-bodied newts (T. marmoratus and T. cristatus superspecies) was also supported. However, the phylogenetic position of the medium-sized newt, T. alpestris could not be clarified.

Adenosine Triphosphatases↗
PubMed1990

The development of the larval pigment patterns in Triturus alpestris and Ambystoma mexicanum.

1. Melanophores and xanthophores are pigment cell derivatives of the NC. In amphibian embryos they migrate from their original position on the neural tube dorsally (into the dorsal fin) as well as laterally (between somites and epidermis) and arrange themselves into typical pigment patterns of the skin. We investigated pigment pattern formation in two species of tailed amphibians, Triturus alpestris (alpine newt) and Ambystoma mexicanum (Mexican axolotl). In larvae of T. alpestris alternating longitudinal stripes or bands of melanophores and xanthophores develop, whereas in larvae of A. mexicanum a barred pattern with alternating transverse bands of melanophores and xanthophores is formed. Iridophores, a third type of pigment cell, are present later in both species and therefore play no role during early larval pigment pattern development. Visibly differentiated melanophores and xanthophores can be distinguished from each other under the light microscope by their contents of black melanins and yellow pterins respectively. With the dopa reaction (indicates tyrosinase in melanophores), and ammonia treatment (stimulates pterin fluorescence in xanthophores), the pigment cell phenotypes can be visualized even before their normal visible differentiation. In the TEM, melanophores and xanthophores can be distinguished from each other by their morphologically distinct pigment organelles and in the SEM by their different surface structure. 2. Because of the NC origin of melanophores and xanthophores and the ease with which these cells can be demonstrated even before they are visible from outside, their different arrangements in Triturus and axolotl embryos offer suitable model systems for studying the migration, interaction and localization of NC derivatives in relation to specific environmental influences. The environment of NC cells are the neural tube, epidermis, somites and lateral plate mesoderm, and the subepidermal ECM, a network of collagen fibrils associated with glycosaminoglycans, proteoglycans and glycoproteins. 3. Development of the pigment pattern in T. alpestris: Melanophores and xanthophores start to leave the NC at stage 28, melanophores slightly earlier than xanthophores. Both cell types become scattered in the dorsolateral trunk. In contrast to melanophores in the axolotl, melanophores in T. alpestris cannot be demonstrated with the dopa reaction before they become visibly black. From stage 29+ onwards, melanophores start to accumulate in zones alongside the dorsal and lateral somite edges, where they form compact stripes later. Xanthophores can be demonstrated from stage 28+ onwards only with the SEM (by means of their specific surface structures) or with the fluorescence microscope (by means of their fluorescing pterins). At state 34, xanthophores become visible externally as yellow cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Ambystoma↗
PubMed1986

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.

Animals↗
PubMed1975

[Evolution of xenografts of embryonic gonads and juvenile ovaries from Ambystoma mexicanum Shaw, transplanted into Triturus alpestris Laur].

Gonad primordium of Ambystoma mexicanum when grafted at tail-bud stage on Triturus alpestris is indefinately tolerated. Testis or ovaries derived from those grafted embryonic gonads follow their differentiaion and reach sexual maturity. Axolotl juvenile ovaries transplanted in Triturus alpestris are also tolerated; the ovocytes achieve the entire vitellogenesis but are not able to be laid.

Ambystoma↗
PubMed1991

Effects of beta-endorphin and naloxone on corticosterone and cortisol release in the newt (Triturus carnifex): studies in vivo and in vitro.

The effects of beta-endorphin and its receptor antagonist, naloxone, on corticosterone and cortisol production in male and female Triturus carnifex were studied in vivo and in vitro. In the in-vivo experiment, the animals were injected s.c. with beta-endorphin and/or naloxone, and killed after 15, 30, 90 and 360 min. In the in-vitro experiment, interrenal tissues, with and without added pituitary, were incubated with beta-endorphin and/or naloxone for 15, 30, 60 and 120 min. The data obtained in vivo and in vitro from males and females were in agreement. Treatment with beta-endorphin caused a significant decrease in corticosterone and cortisol release, while naloxone induced an increase in the two corticosteroids at the same times as the decrease caused by beta-endorphin. The combined beta-endorphin plus naloxone treatment did not change corticosterone and cortisol levels. These results suggest that, in Triturus carnifex, opioids are involved in the regulation of the hypothalamo-pituitary-interrenal axis. In particular, the in-vitro results indicate a direct effect of opioids on interrenal steroidogenesis.

Animals↗
PubMed1988

The monoaminergic system in the diencephalon of the newt tadpole, Triturus alpestris (Mert). A histofluorescence study.

The distribution of monoamines in the brain of Triturus alpestris tadpoles was investigated with the Falck-Hillarp histofluorescence method. This study concerns especially the diencephalon where is observed the major part of the monoaminergic (MA) neurons. MA fiber tracts are found in the striatum, the septal nucleus and the medial and lateral forebrain bundles of the telencephalon. In the anterior diencephalon cerebrospinal fluid (CSF)--contacting catecholaminergic cells are localized within the preoptic recess organ (PRO) and fibers in the preoptic nucleus. More caudally both CSF-contacting catecholaminergic and serotonergic cells can be demonstrated in the paraventricular organ (PVO) and in the nucleus infundibularis dorsalis (NID). In the hypothalamo-hypophysial tract, numerous catecholaminergic (CA) fibers occur in the median eminence and in the pars intermedia. The origin of these fibers is not elucidated but a participation of the PRO, PVO and NID seems possible. CA cells without contact with the CSF are localized laterally to the PVO, PVO-accompanying cells, and in the nucleus reticularis mesencephali (NRM). The distribution of MA neurons in the brain of Triturus alpestris appears more similar to that of anurans than that of the primitive urodeles.

Animals↗
PubMed1996

Testicular cell cytoskeleton in the newt, Triturus marmoratus marmoratus, during the annual cycle.

Light and electron microscopy immunohistochemical studies and Western blotting analysis of cytoskeletal proteins have been carried out in the testis of the marbled newt (Triturus marmoratus marmoratus) during the annual testicular cycle. The present findings revealed homologies and differences with regard to those reported in the testes of mammals and other vertebrates. Changes in immunohistochemical expression have also been detected in the course of the annual cycle. Actin and tubulin, which were scanty and diffusely located in spermatogonia and spermatocytes, increased their expression and reorganized during spermiogenesis. Vimentin and keratin, undetected in spermatogonia and spermatocytes, were expressed in differentiating spermatids and spermatozoa. In these cells, actin might be related with the connection of the axial fiber to the undulating membrane and the coordination of movement by both structures, while vimentin might be involved in the maintenance of the spatial relationship between the axoneme and the marginal fiber. During the first stages of spermatogenesis, the cytoplasm of Sertoli cells (follicular cells) showed a diffuse immunoreaction to actin, myosin, and tubulin and no vimentin immunolabeling. In advanced spermiogenesis, the follicular cells showed an intense immunoreaction to actin, myosin, tubulin, and vimentin in the apical projections that surrounded the spermatid heads. These apical cytoskeletal components might be involved in spermatid elongation, since the spermatids display no manchette, and in spermatozoon positioning and grouping. The colocalization of myosin and actin in the follicular cells suggests that actin filaments from contractile bundles and that contraction might be involved in changes in the Sertoli cell shape that accompany germ cell development during spermatogenesis. The interstitial cells immunostained to actin, myosin, tubulin, and vimentin. These cells, together with follicular cells, seemed to form the glandular tissue cells which showed a similar immunophenotype. The cells that surrounded the efferent duct epithelium immunostained to desmin, and they are probably contractile cells involved in sperm evacuation.

Activity Cycles↗
PubMed1999

Sperm storage in females of the smooth newt (Triturus v. vulgaris L.): I. Ultrastructure of the spermathecae during the breeding season.

Sperm storage in cloacal spermathecae was studied in females of Triturus v. vulgaris collected early in the breeding season in southern England. Females collected in terrestrial situations, presumably unmated, were mated in the laboratory, and the ultrastructure of the transferred sperm and the spermathecae was observed at various intervals after mating. Sperm from a spermatophore cap lodged in a female's cloacal orifice can migrate into spermathecae within 1 hr after mating. Spherical structures on the axial fibers of some sperm in the cap could indicate immaturity. Disorderly clusters of sperm from the cap are still present in the cloacal chamber 12 hr after mating but are absent 24 hr after mating. During storage, sperm often are in tangled masses in the spermathecal tubules. The sperm are coated with spermathecal secretions, and some sperm nuclei were observed embedded in the spermathecal epithelium. Little evidence for spermiophagy early in the breeding season was found. During oviposition, mazes of sperm occur external to the spermathecal orifices, and sperm may be released in this condition onto eggs as they pass through the cloaca. The tangled clusters in which sperm are found from pick-up to oviposition are hypothesized as an adaptation to reduce the effectiveness of sperm competition from the ejaculates of rival males. Additional studies, using the same protocol and covering the entire cycle of sperm storage, are necessary to enable interspecific comparisons leading to phylogenetic hypotheses.

Adaptation, Physiological↗
PubMed1999

Ultrastructure and lectin cytochemistry of the cloacal pelvic glands in the male newt Triturus marmoratus marmoratus.

The cloacal organ of Salamandridae species contains four glands: pelvic, dorsal, ventral, and Kingsbury's glands. Pelvic glands have been studied only by light microscopy with conventional methods, and consist of multiple tubular serous glands with a prismatic epithelium which contains numerous PAS positive secretory granules. The present report is an ultrastructural and lectin cytochemistry characterization of the pelvic glands of Triturus marmoratus marmoratus throughout the reproductive cycle. Our methods consisted of conventional electron microscopy, and colloidal-gold lectin cytochemistry of the following lectins: WGA, ConA, LcA, UEA-I, PNA, SBA, and HPA. In the prereproductive period, the glands showed a tall epithelium which consisted of two cell types, dark and clear cells, surrounded by elongated, myoepithelial cells. Both dark and clear cells showed the ultrastructural characteristics of secretory cells, and exhibited many secretory granules in the apical cytoplasm. Areas showing densely packed, degenerating cell organelles--which were not surrounded by membrane--were observed in the dark cells whereas the clear cells showed large heterolysosomes. In the postreproductive period the number of secretory granules decreased, the rough endoplasmic reticulum was less developed, and areas of degenerating organelles were absent. In addition, small basal cells appeared. The results of the lectin histochemistry study were similar in both reproductive periods. In the epithelial cells, the rough endoplasmic reticulum, the Golgi complex, and secretory granules exclusively labeled to ConA. In all cell types, the nuclei reacted to all lectins while the cytosol only reacted to LcA lectin. The ultrastructural and histochemical characteristics of the pelvic glands of T. marmoratus suggest that these glands could be homologous to the mammalian seminal vesicles and prostate.

Animals↗
PubMed1976

Electron microscopic stldy of the developing lateral-line organ in the embryo of the newt, Triturus pyrrhogaster.

The morphological sequence of events occurring in the development of the lateral-line organ was investigated in the embryo of the newt, Triturus pyrrhogaster, by means of electron microscopy. Six stages of development have been defined for convenience of description. The primordium of the lateral-line organ migrating from the pre- and post-auditory placodes is segmented into a small cell mass accompanied by a few Schwann cells enveloping nerve fibers. In the clump of cells of the lateral-line primordium, three kinds of cells are already distinguished by different degrees of darkness, i.e., dark, light and slightly dark cells (stage I). Both clump and Schwann cells enter the epidermis together (stage II). The two groups of cells are separated by the inner layer of the epidermis (stage III). The organ-forming cells elongate and the inner layer of the epidermis separates the Schwann cells from the clump (stage IV). The apexes of organ-forming cells are exposed out of the epidermis (stage V). The lateral-line organ is almost matured. The dark cells correspond to type II of the supporting cells, the light cells correspond to type I of the supporting cells and the slightly dark cells correspond to the receptor cells because of the existence of cilia and a cuticular plate. The afferent nerve endings are found on the basal surface of the receptor cell (stage VI).

Animals↗
PubMed1987

Effect of optic nerve denervation on lens regeneration in adult newts, Triturus viridescens.

This experiment was designed to study the effect of optic nerve denervation on lens regeneration. Adult newts, Triturus viridescens, were divided into 4 groups: A) lentectomized newts, which served as controls, B) lentectomized newts with the optic nerve exposed and lightly palpated, C) newts with lens removed and the optic nerve cut simultaneously, and D) newts with lens removed first and optic nerve cut 7 days and 14 days later. Control animals required about 21 days to produce a new lens, and animals in group B demonstrated the same regeneration rate. In groups C and D, the regeneration rate was greatly retarded during the first 14 days. However, most of these animals regenerated a perfect new lens by 21 days postlentectomy.

Animals↗
PubMed2002

Pituitary adenylate cyclase-activating polypeptide induces testicular testosterone synthesis through PGE(2) mediation in crested newt, Triturus carnifex.

The aim of the present work was to study the possible role of adenylate cyclase-activating polypeptide (PACAP) 38 in the testicular intracellular mechanism regulating steroidogenesis of crested newt, Triturus carnifex. Gonads were incubated in vitro with PACAP 38 and prostaglandin (PG) E(2) alone or with inhibitors of cyclooxygenase (COX), adenylate cyclase (AC), and phospholipase C (PLC) for 30 min and 60 min. PGE(2), PGF(2 alpha), testosterone, and estradiol-17 beta were measured in the culture medium; aromatase (AR) activity and cAMP were assessed in the tissue. PACAP 38 increased PGE(2) (30 min and 60 min), estradiol-17 beta (60 min), cAMP (60 min), and AR (60 min) but decreased testosterone (60 min). PGE(2) increased estradiol-17 beta, cAMP, and AR and decreased testosterone at 30 and 60 min.PLC inhibitor counteracted the effects of PACAP 38, while AC inhibitor counteracted these effects except for PGE(2) increase. AC inhibitor counteracted the effects of PGE(2), while PLC did not. COX inhibitor decreased PGF(2 alpha) (30 min and 60 min), PGE(2) (30 min and 60 min), estradiol-17 beta (60 min), cAMP (60 min), and AR (60 min), but increased testosterone (60 min). These in vitro results suggest that, in newt testis, PACAP 38 acts on PLC, inducing the increase of PGE(2) which, in turn, acting on AC, increases AR activity with the consequent estradiol-17 beta increase and testosterone decrease.

Animals↗
PubMed1975

Regeneration of the retina in the adult newt, Triturus cristatus, following surgical division of the eye by a limbal incision.

The regeneration of a retina in adult Triturus cristatus, following surgical division of the eye by a limbal incision, was studied. In agreement with recent reports, it was found that the regenerating retina is dervied from two sources; the retinal pigment epithelium and the pars ciliaris retinae. However, following a limbal incision, most of the retina appears to be derived from the retinal pigment epithelium in the posterior part of the eye. An unexpected finding of this study was that large cystlike spaces form in the fundal regions of the eye, between the regenerating retina and the retinalpigment epithelium. These spaces appear between five and eight days post-operative and persist long enough (25 to 30 days postoperative) to disrupt the fundal portion of the rengenerating retina and to cause it to lag behind the rest of the regenerate, in its development. The relationship of these observations to the genesis of positional markers in the regenerating retina is discussed.

Animals↗
PubMed1975

Non-glial phagocytes within the degenerating optic nerve of the newt (Triturus viridescens).

Two non-glial phagocytes were found to participate along with ependymoglial cells in Wallerian degeneration of the severed optic nerve of the newt (Triturus viridescens). The first type of non-glial cell (polymorphonuclear phagocyte) was positively identified as a neutrophil and participates in the early stages of degeneration. Cells of this type make a brief appearance, reaching a peak by the second postoperative day (2 p.o.d.), and quickly diminish until few can be found by 4 p.o.d. Neutrophils invade the degenerating optic nerve from surrounding connective tissue spaces, most likely, through channels which penetrate the nerve parenchyma. The second type of non-glial cell is an invading mononuclear phagocyte which exhibits characteristics of microglial cells reported in other vertebrate species. Such cells appear in the nerve much later than the neutrophils and towards the end of Wallerian degeneration (6-10 p.o.d.). Their mode of entry and exit appears to be the same as that reported for neutrophils. The neutrophils and microglial-like, mononuclear phagocytes may serve to supplement the histolytic action of the ependymoglial cells, picking up scattered fragments of degenerating myelin and axons.

Animals↗
PubMed1977

The effect of nerve growth factor and limb regeneration on the spinal and sympathetic ganglia of Triturus.

In Triturus administration of NGF causes hypertrophy of neuronal perikarya of third dorsal root ganglia and post-subclavian sympathetic ganglia. Two classes of neurons were characterized cytologically in the third dorsal root ganglia. Some evidence is presented which indicates that Class I neurons respond more vigorously to NGF. It is suggested that Class I neurons are similar to the medio-dorsal neuroblasts of chick embryos in their response to NGF. Forelimb amputation did not cause neuronal hypertrophy in either third dorsal root ganglia or post-subclavian sympathetic ganglia. The incorporation of (3H)-uridine increases to a highly significant extent in third dorsal root ganglia following NGF treatment or limb amputation. When NGF treatment and limb amputation were combined, no further increase in (3H)-uridine incorporation was noted. This suggests that RNA synthesis is maximally stimulated by either NGF treatment alone or limb amputation alone.

Amputation, Surgical↗