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PubMed2000

Three new helminth species from two endemic plethodontid salamanders, Typhlomolge rathbuni and Eurycea nana, in central Texas.

Helminthological examination of two rare, endemic species of plethodontid salamanders, the Texas blind salamander (Typhlomolge rathbuni Stejneger) and the San Marcos dwarf salamander (Eurycea nana Bishop), from the subterranean waters and springs in San Marcos, Hays County, central Texas, USA revealed the presence of three new, previously undescribed species of intestinal helminths: Brachycoelium longleyi sp. n. (Trematoda) from T. rathbuni (type host) and E. nana, Dendronucleata americana sp. n. (Acanthocephala) from T. rathbuni, and Amphibiocapillaria texensis sp. n. (Nematoda) from T. rathbuni; nematode larvae probably belonging to the last named species were recorded from E. nana. Brachycoelium longleyi can be distinguished from all congeners primarily by its conspicuously small eggs among other features, whereas A. texensis differs from its closest congeneric species A. tritonispiunctati mainly in the structure of mature eggs and a markedly shorter spicule. Dendronucleata americana is the first species of the family Dendronucleatidae from the New World, differing from its Asian congeners mainly in the number and arrangement of proboscis hooks, number of giant hypodermic nuclei and in the position of testes.

Acanthocephala↗
PubMed2005

Ontogenic delays in effects of nitrite exposure on tiger salamanders (Ambystoma tigrinum tigrinum) and wood frogs (Rana sylvatica).

Under certain conditions, nitrite can be present in freshwater systems in quantities that are toxic to the fauna. I exposed wood frog (Rana sylvatica) and eastern tiger salamander (Ambystoma tigrinum tigrinum) embryos and young tadpoles and larvae to elevated concentrations of nitrite in chronic toxicity tests: 0, 0.3, 0.6, 1.2, 2.1, 4.6, and 6.1 mg/L NO2-N, exposing individuals as both embryos and larvae. Nitrite caused significant declines in wood frog hatching success (3.4 mg/L NO2-N, wood frog), and lower concentrations caused significant mortality during the early larval stages (4.6 mg/L NO2-N, salamander; 0.5 mg/L NO2-N, wood frog). Later tests exposing individuals to nitrite only after hatching showed that both wood frog and tiger salamander vulnerability to nitrite declined shortly after hatching. Hence, examining a single life-history stage, especially later in development, may miss critical toxic effects on organisms, causing the researcher potentially to underestimate seriously the ecological consequences of nitrite exposure.

Ambystoma↗
PubMed1992

An additional discovery of salamanders, Salamandrella keyserlingii Dybowski, with no blood vessels in the brain parenchyma except in the olfactory bulb.

The brain parenchyma of Salamandrella keyserlingii Dybowski (SKD) is not vascularized except in the olfactory bulb. In the brains of SKD, neuronal and glial perikarya are seen densely aggregated at the periventricular regions and no blood vessels enter the brain parenchyma from the meningeal vessels. Former investigators have discovered no blood vessels in the brains of nine species of salamanders in the Hynobiidae, all of which inhabit Japan. This report adds one more avascular species of salamander which is found in Hokkaido, Japan. The reasons why the brain of Hynobiid salamanders is not vascularized and why only the olfactory bulbs are vascularized cannot be speculated on at the present time.

Animals↗
PubMed2000

Cloning of a cDNA encoding cathepsin D from salamander, Hynobius leechii, and its expression in the limb regenerates.

Cathepsin D is a major lysosomal aspartic proteinase participating in the degradation or modification of intra- and extracellular matrix molecules, and its activity is known to increase in the process of tissue reorganization during the early phase of salamander limb regeneration. Here, we report the cloning of a salamander cathepsin D cDNA from Hynobius leechii and its expression profile in normal and retinoic acid (RA) treated limb regenerates. The gene expression of cathepsin D increased notably during the dedifferentiation stage and decreased gradually thereafter. Furthermore, RA that enhances dedifferentiation of regenerating salamander limb caused the elevation of cathepsin D expression both in terms of level and duration. These results suggest that cathepsin D plays important role(s) in the dedifferentiation process, and enhancement of cathepsin D expression might be closely related to RA-evoked pattern duplication.

Amino Acid Sequence↗
PubMed2005

Evaluation of two methods for measuring nonspecific immunity in tiger salamanders (Ambystoma tigrinum).

Study of amphibian immunotoxicology is a growing area of research, but very little information is available on how environmental contaminants affect disease resistance in urodele amphibians. Urodele amphibians lack the more highly evolved aspects of the specific immune system that are present in anurans, birds, and mammals. Instead, these animals rely more heavily on innate defense mechanisms than do anurans to provide rapid, nonspecific protection from pathogens. Thus, it is prudent that immunotoxicologic research with urodele amphibians includes an evaluation of effects of contaminant exposure on nonspecific immunity. The objectives of this study were to measure the phagocytic and oxidative-burst activity of peritoneal neutrophils collected from a urodele, the tiger salamander (Ambystoma tigrinum), and to evaluate the use of these assays in immunotoxicologic research using urodele amphibians. Using tiger salamanders collected in August 2000, phagocytosis and oxidative-burst assays modified from mammalian protocols were conducted through October 2001. Results indicated that large numbers of peritoneal neutrophils for use in immunotoxicologic tests can be collected from salamanders injected with thioglycollate. Moreover, these neutrophils readily engulfed foreign material (phagocytic activity) and produced measurable amounts of hydrogen peroxide (oxidative-burst activity). Phagocytosis was effectively inhibited by incubating cells with sodium azide (P<0.001), and quantification of phagocytosis using flow cytometry was well correlated with manual counts (r=0.84, P<0.001). Dexamethasone treatment reduced phagocytic activity as measured by manual counts (P<0.02), suggesting that this test is useful for detecting alteration by immunosuppressive agents. In contrast, oxidative function was unaffected by dexamethasone treatment, and results from the oxidative-burst assay were generally less consistent than those from the phagocytosis assay. Based on these results, phagocytic activity of peritoneal neutrophils may be a useful endpoint in immunotoxicologic studies to evaluate the impact of environmental contaminants on innate defense mechanisms in urodele amphibians.

Ambystoma↗
PubMed2000

Cloning and functional characterization of salamander rod and cone arrestins.

PURPOSE: To clone, localize, and determine functional binding characteristics of rod and cone arrestins from the retina of the tiger salamander (Ambystoma tigrinum). METHODS: Two arrestins from salamander retina were cloned on the basis of their homology to known arrestins from other species. The expression pattern of these arrestins (SalArr1 and SalArr2) in the retina was determined by immunocytochemistry and in situ hybridization. SalArr1 and SalArr2 were expressed and functionally characterized. RESULTS: Both immunocytochemistry and in situ hybridization show that SalArr1 and SalArr2 localized specifically to rod and cone photoreceptors, respectively. SalArr1 demonstrated a characteristic high selectivity for light-activated phosphorylated rhodopsin (P-Rh*) and significant species selectivity, binding preferentially to amphibian rhodopsin over bovine rhodopsin. Mutant constitutively active forms of SalArr1 demonstrated a 2- to 4-fold increase in P-Rh* binding (compared with wild-type protein) and an even more dramatic (up to 25-fold) increase in binding to unphosphorylated Rh* and dark P-Rh. Constitutively active SalArr1 mutants also showed a reduced specificity for amphibian rhodopsin. The ability of Escherichia coli-expressed SalArr1, SalArr2, and an SalArr1-3A (L369A,V370A,F371A) mutant to bind to frog Rh* and P-Rh* and to compete with tritiated SalArr1 for amphibian P-Rh* was compared. SalArr1 and its mutant form bound to amphibian P-Rh* with high affinity (Ki = 179 and 74 nM, respectively), whereas the affinity of SalArr2 for P-Rh* was substantially lower (Ki = 9.1 microM). CONCLUSIONS: SalArr1 and SalArr2 are salamander rod and cone arrestins, respectively. Crucial regulatory elements in SalArr1 are conserved and play functional roles similar to those of their counterparts in bovine rod arrestin. Rod and cone arrestins are relatively specific for their respective receptors.

Ambystoma↗
PubMed2000

Antibacterial and hemolytic activity of the skin of the terrestrial salamander, Plethodon cinereus.

As resistance increases against fungal antibiotics, antimicrobial peptides are receiving attention as possible replacements. The dermal glands of frogs secrete, among other things, antimicrobial peptides. As part of the innate immune system, stressors may affect the production of antimicrobial peptides by dermal glands. The dermal secretions of some salamanders have been examined for their toxic secretions, but little attention has been given to salamander antimicrobial peptides. This study examines the skin from the tail region for the production of antimicrobial peptides in the terrestrial salamander, Plethodon cinereus. Fractions of tail extracts were isolated using cation-exchange chromatography and reverse-phase HPLC. An HPLC fraction eluting at 15.75 min (HPLC run: 30 min, 30-80% acetonitrile/water gradient, Aquapore RP-300 C18 column) showed activity against Staphylococcus aureus but not against Escherichia coli. The antibacterial activity gradually increased over a 4-hr incubation time up to about 85% inhibition of bacterial growth. Lysis of guinea pig red blood cells also increased gradually over a 1-hr time period. J. Exp. Zool. 287:340-345, 2000.

Animals↗
PubMed1979

Light and scanning microscopy of the taste organs and vascularization of the tongue of the spotted salamander, Salamandra salamandra (L.).

The mucosa of the spotted salamander tongue and its taste organs were investigated by means of light and scanning electron microscopy. The most striking feature of the salamander tongue is an almost complete lack of papillae which are replaced by long, radially disposed folds with linear arrays of taste organs along their ridges. In respect of morphology, the taste organs of the salamander occupy an intermediate position between the taste buds of Urodela and taste discs of Salientia. Scanning electron microscopic examination of microcorrosion casts of the blood vessels of the tongue has revealed that the structure of subepidermal capillary network reflects the topography of the tongue surface and the distribution of its taste organs. In the core regions of the folds the capillary loops accompanying gustatory receptors empty via their shorter, descending arms into the draining vessels, the initial segments of which retain a course parallel to that of the folds. In the few fungiform papillae the capillary vessels form single loops whose distal ends come to lie in the vicinity of taste discs.

Animals↗
PubMed1996

Similarities and differences in the cytoarchitecture of the tectum of frogs and salamanders.

Frogs exhibit a morphologically complex (multiply laminated) optic tectum, while salamanders have one of the morphologically simplest tecta among vertebrates. In a comparative approach, the morphology of tectal projection neurons is investigated in three salamander species, Hydromantes italicus, H. genei and Plethodon jordani, and two frog species, Discoglossus pictus and Eleutherodactylus coqui, by means of retrograde Biocytin labeling complemented by intracellular Biocytin staining of cells. Despite striking differences in the gross anatomy of the tectum, salamanders and frogs have the same types of tectal neurons with respect to their dendritic arborization and the pattern of ipsilaterally and bilaterally ascending (to praetectum and thalamus) and ipsilaterally or contralaterally descending projections (to nucleus isthmi, medulla oblongata and rostral spinal cord). In the light of these findings, the relationship between morphological complexity of the tectum and behavioral complexity (feeding behavior) is discussed.

Animals↗
PubMed1998

Mechanics of lung ventilation in a larval salamander Ambystoma tigrinum.

The larval stage of the tiger salamander Ambystoma tigrinum is entirely aquatic, but the larvae rely on their lungs for a large proportion of their oxygen uptake. X-ray video and pressure measurements from the buccal and body cavities demonstrate that the larvae inspire using a two-stroke buccal pump and exhale actively by contracting the hypaxial musculature to increase body pressure. Larvae begin a breath by expanding the buccal cavity to draw in air through the mouth, while simultaneously exhaling air from the lungs to mix with the fresh air in the buccal cavity. The mouth then closes, and the buccal cavity compresses to pump a portion of the mixture into the lungs. The remaining air in the buccal cavity is then released as bubbles from the mouth and gill slits. Ventilatory volumes estimated from X-ray video records indicate that approximately 80 % of the air pumped into the lungs is fresh air and 20 % is previously expired air. Exhalation in larval tiger salamanders is active, powered by contraction of all four layers of lateral hypaxial musculature. Electromyography indicates that the transverse abdominis (TA) muscle is active for the longest duration and shows the highest-amplitude activity, but the external oblique superficialis, the external oblique profundus and the internal oblique also show consistent, low-level activity. The finding that the TA muscle is active during exhalation in larval tiger salamanders contributes to a growing body of evidence that the use of the TA for exhalation is a primitive character for tetrapods.

Journal Article↗
PubMed1998

Mechanics of lung ventilation in a larval salamander Ambystoma tigrinum.

The larval stage of the tiger salamander Ambystoma tigrinum is entirely aquatic, but the larvae rely on their lungs for a large proportion of their oxygen uptake. X-ray video and pressure measurements from the buccal and body cavities demonstrate that the larvae inspire using a two-stroke buccal pump and exhale actively by contracting the hypaxial musculature to increase body pressure. Larvae begin a breath by expanding the buccal cavity to draw in air through the mouth, while simultaneously exhaling air from the lungs to mix with the fresh air in the buccal cavity. The mouth then closes, and the buccal cavity compresses to pump a portion of the mixture into the lungs. The remaining air in the buccal cavity is then released as bubbles from the mouth and gill slits. Ventilatory volumes estimated from X-ray video records indicate that approximately 80% of the air pumped into the lungs is fresh air and 20% is previously expired air. Exhalation in larval tiger salamanders is active, powered by contraction of all four layers of lateral hypaxial musculature. Electromyography indicates that the transverse abdominis (TA) muscle is active for the longest duration and shows the highest-amplitude activity, but the external oblique superficialis, the external oblique profundus and the internal oblique also show consistent, low-level activity. The finding that the TA muscle is active during exhalation in larval tiger salamanders contributes to a growing body of evidence that the use of the TA for exhalation is a primitive character for tetrapods.

Ambystoma↗
PubMedDate not supplied

Comparative biology of sperm storage in female salamanders.

Females in seven of the ten families of salamanders possess cloacal glands called spermathecae that store sperm. The annual cycle of sperm storage has been studied by light and electron microscopy in eight species representing five families. In these taxa, we recognized 14 characters associated with the spermathecae and traced their evolution on a phylogeny of salamanders based upon other characters. The plasticity and phyletic significance of the spermathecal characters varied greatly. Plethodontids have complex spermathecae while other families possess simple spermathecae; thus, this character has phyletic value as well as being highly conserved within the Salamandroidea. Other characters, such as carbohydrate histochemistry, are highly plastic and show no obvious phyletic trends. The significance of some of these variable characters, such as duration of sperm storage, is apparent only after including in the analysis other aspects of the reproductive cycle, such as length of the mating season. Additional comparative studies, employing the protocol used in this paper, will help further clarify the relationships between phyletic and functional variability in sperm storage mechanisms in salamanders.

Animals↗
PubMed2026

Identification and characterization of the HSP gene family in the Chinese giant salamander: Expression patterns under combined environmental stress.

BACKGROUND: The Chinese giant salamander (Andrias davidianus) is a critically endangered living fossil species that is highly sensitive to changes in water temperature. However, systematic studies on the heat shock protein (HSP) gene family and its response mechanisms to environmental stress in this species remain limited. This study utilized transcriptome data from captive-bred salamanders exposed to combined temperature and pathogen stress. Bioinformatics tools were employed to identify the HSP gene family of A. davidianus (AndHSP) and to analyze their evolution, structure, and function, thereby revealing their regulatory mechanisms in response to environmental stress. RESULTS: A total of 72 AndHSPs were identified and classified into five subfamilies. Phylogenetic analysis revealed that each subfamily is evolutionarily conserved and functionally related. Gene expression analysis demonstrated that pathogen infection induced the expression of AndHSPs, and elevated temperature significantly intensified this response. Nine key differentially expressed genes were identified, predominantly from the AndHSP70 subfamily, with AndHSP70-18 exhibiting rapid heat-induced expression. Tissue-specific analysis showed high expression of AndHSP60 in the spleen. A qPCR validation confirmed the reliability of the transcriptome expression results. CONCLUSIONS: This study presents the first systematic identification of the AndHSP gene family and elucidates its cooperative stress response mechanisms under combined temperature and pathogen stress. These findings provide a molecular basis for understanding the species' environmental adaptation and have important implications for its conservation and artificial breeding.

Animals↗
PubMed1999

Morphology, axonal projection pattern, and response types of tectal neurons in plethodontid salamanders. I: tracer study of projection neurons and their pathways.

In three salamander species (Hydromantes italicus, H. genei, Plethodon jordani), the tectobulbospinal and tectothalamic pathways and their cells of origin were studied by means of anterograde and retrograde biocytin and tetramethylrhodamine tracing. In plethodontid salamanders, five types of tectal projection neurons were identified. TO1 neurons have widefield dendritic trees that arborize in the layers of retinal afferents and form a neuropil in the superficial layer; axons constitute the crossed tectospinal tract. Dendrites of TO2 cells have the largest dendritic trees that arborize in the intermediate and deep layers of retinal afferents; axons constitute a lateral uncrossed tectospinal tract. TO3 cells have widefield dendritic trees that arborize in the deep layer of retinal afferents and in the layer of tectal efferents; axons constitute a superficial uncrossed tectospinal tract. TO4 cells have slender primary dendrites and small-field dendritic trees that arborize in the intermediate layers of retinal afferents; axons constitute another lateral uncrossed tectospinal tract. TO2, TO3, and TO4 cells also have ascending axons that run to the ventral and dorsal thalamus. TO5 cells have slender primary dendrites and small-field dendritic trees that extend into the superficial layers of retinal afferents; their fine axons constitute the bulk of the pathways ascending to the ipsilateral and contralateral thalamus. These morphological types of projection neurons and their ascending and descending axonal pathways closely resemble those found in frogs, reptiles, and birds. Their role in visual and visuomotor functions is discussed.

Animals↗
PubMed1998

Primary and secondary somatosensory projections in direct-developing plethodontid salamanders.

In three species of plethodontid salamanders (Plethodon jordani, Hydromantes italicus, and Bolitoglossa subpalmata), primary and secondary somatosensory pathways were investigated by means of tract-tracing in vivo and in vitro using biocytin, horseradish peroxidase, and neurobiotin. Afferent sensory fibers of cranial nerves V, VII, and X and the brachial nerve run in the dorsal funiculus of the medulla oblongata and spinal cord. Fibers ascend to the level of, but do not enter, the cerebellum. In the caudal medulla oblongata, sensory tracts of the cranial nerves descend in a dorsal and a dorsolateral bundle and reach the level of the fourth spinal nerve. Two bundles are likewise formed by spinal afferent fibers, which descend to the level of the seventh spinal nerve. Secondary somatosensory projections ascend in contralateral ventral, contralateral lateral, and ipsilateral lateral tracts, the latter two corresponding to the spinal lemniscal tracts of Herrick. These tracts reach the cerebellum, mesencephalic, and diencephalic targets (tegmentum, torus, tectum, tuberculum posterius, pretectum, and ventral thalamus) ipsi- and contra-laterally. The projection to the tectum is confined to fiber layer 4. Fibers of the ascending tracts cross in the cerebellar and tectal commissure. Our study demonstrates that the ascending secondary somatosensory pathways of plethodontid salamanders differ remarkably from those of other amphibians.

Animals↗
PubMed2000

Morphology, axonal projection pattern, and responses to optic nerve stimulation of thalamic neurons in the salamander Plethodon jordani.

In the salamander Plethodon jordani, the morphology and axonal projections of thalamic (TH) neurons and their responses to electrical optic nerve stimulation were determined by intracellular recording and biocytin labeling under in vitro, whole-brain conditions. Based on their axonal projections, labeled neurons (n = 76) were divided into the following groups: TH1 neurons, with mostly ipsilateral projections to the striatum; TH2 neurons, with ipsilateral or bilateral projections to the medial amygdala and nucleus accumbens; TH3 neurons, with bilateral projections to the medial and dorsal pallium; TH4 neurons, with mostly ipsilateral projections to the striatum and ipsilateral projections to the tectum opticum, tegmentum, and rostral medulla oblongata; and TH5 neurons, with ipsilateral projections to the tegmentum, medulla oblongata, and rostral spinal cord without (TH5.1) or with (TH5.2) additional projections to the optic tectum. TH1-TH4 neurons are found in the dorsal thalamus and around the sulcus medialis, and TH5 neurons are found in the ventral thalamus. Labeled neurons with ascending projections, i.e., the more dorsally situated TH1-TH4 neurons, are mostly inhibited by electrical stimulation of the optic nerve and have significantly longer latencies (mean +/- S.D., 42.1 +/- 11.6 msec) than neurons with exclusively descending projections, i.e., the ventrally located TH5 neurons (8.5 +/- 6.1 msec), which receive the bulk of retinal afferents and show excitation at electrical optic nerve stimulation. Neurons recorded without labeling in the dorsal thalamus likewise exhibit mostly inhibition and have significantly longer latencies (35.7 +/- 18.9 msec) than those recorded in the ventral thalamus (10.9 +/- 7.7 msec), which mostly show excitation. None of the neurons recorded in the dorsal thalamus followed repetitive stimulation of the optic nerve. Thus, neurons situated in the dorsal thalamus and projecting to pallial or subpallial telencephalic targets are unlikely to receive monosynaptic or oligosynaptic, excitatory retinal input. Accordingly, no retino-thalamo-telencephalic pathway homologous to that found in amniotes appears to exist in salamanders.

Action Potentials↗
PubMed2000

Ultrastructure of the skin mechanoreceptors of the Chinese giant salamander, Andrias davidianus.

The ultrastructure of two kinds of mechanoreceptive organs, pit organs and neuromasts, in the skin of adult giant salamanders (Andrias davidianus) was studied by transmission electron microscopy. Neuromasts and pit organs differ in their types of synapses, the spatial distribution of kinocilia on sensory cells, and in the degree to which sensory cells are separated by processes of the supporting cells; the two organs probably serve complementary functions. The neuromasts in A. davidianus differ from those of other salamanders in the orientation of kinocilia, in the extent of intrusion of supporting cells into the sensory layer, in the degree of thickening of the synaptic membranes, in the distribution of synaptic spheres, and by the absence of a cupula.

Animals↗
PubMed1994

Pattern duplication by retinoic acid treatment in the regenerating limbs of Korean salamander larvae, Hynobius leechii, correlates well with the extent of dedifferentiation.

In the regenerating limbs of Korean salamanders, Hynobius leechii, retinoic acid (RA) induces duplication of skeletal structures in the proximodistal (PD) axis and often in the transverse axes. In the present study, the stage-dependent effects of RA for the duplication of limb skeletal structures at two amputation levels, the distal stylopodium and the distal zeugopodium, were studied using larval limbs of Korean salamanders. The results showed that the mean level of proximalization (MLP) by RA treatment increased during the stages of dedifferentiation and early bud formation while the MLP declined thereafter in both amputation levels. The decline of the MLP at the later stages of regeneration was due to the high frequency of hypomorphic regeneration or blocked regeneration. When the effects of RA treatment at two amputation levels were compared, the overall trends were similar but the actual timing was delayed for 2-4 days in the proximal level of amputation. Furthermore, the peak level of proximalization was achieved earlier and the peak level remained longer in the distal stylopodial level of amputation compared to the distal zeugopodial level of amputation. Since the histological observations revealed that the dedifferentiation period was also extended up to 2-4 days in the proximal level of amputation, the acid phosphatase activity during the course of regeneration was measured to look for a quantitative relationship between the enzyme activity and the states of dedifferentiation. The results show that the level and the duration of acid phosphatase activity in the upper arm regenerates are both higher and longer than those in the lower arm regenerates. Furthermore, RA treatment caused an increase in acid phosphatase activity. Thus our results suggest that the state of dedifferentiation might be closely linked to the extent of proximalization of regenerating limbs by RA treatment.

Acid Phosphatase↗