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PubMed1995

The interactive effects of low pH, toxic metals, and DOC on a simulated temporary pond community.

The effects of pH, potentially toxic metals, and dissolved organic carbon (DOC) on a freshwater, palustrine wetland assemblage (three amphibian species, one mosquito genus, and 18 genera of algae) were examined in 500 liter outdoor mesocosms. The design was fully factorial and exposure levels were set to represent concentrations that occur naturally in temporary ponds in central Pennsylvania, an acid-impacted region of the USA. Water chemistry was precisely regulated by use of an artificial pond water rather than using stream water or well water with partially unknown composition. Jefferson (Ambystoma jeffersonianum) and spotted salamanders (A. maculatum) were negatively impacted by low pH and high metal concentrations. Wood frogs (Rana sylvatica) were negatively impacted by high DOC and low pH. These negative effects included increased time to metamorphosis and decreased survival and wet body mass at metamorphosis. Mosquito emergence was unaffected by pH, metals, or DOC. All three abiotic factors (metals, pH, and DOC) interacted to determine algal taxonomic composition. Hence, abiotic conditions may determine amphibian reproductive success both directly and indirectly, via effects on algae, since a primary food source of salamanders, zooplankton, are affected by changes in algal composition. These complex interactions should be considered in evaluating the possible toxicity of pond waters to fauna and flora. Until now, pH alone has been considered the primary limiting factor for amphibians breeding in temporary ponds.

Journal Article↗
PubMed1984

A resonance Raman study of Ambystoma tigrinum hemoglobins: evidence for intraspecies hemepocket variations.

Using resonance Raman difference spectroscopy, the Raman-active vibrational modes of hemoglobins from adult, neotenic, and larval forms of the salamander, Ambystoma tigrinum have been compared to each other and to human hemoglobin. The local heme environment of the adult and neotenic proteins were identical and differed from that of the larval protein. Differences were observed in modes sensitive to porphyrin pi electron density and axial ligation. Systematic differences were also observed between human and adult salamander hemoglobins particularly in modes sensitive to the heme vinyl environment. The relationship between these environmental differences, oxygen binding affinity, and the effects of allosteric modulators are discussed.

Aging↗
PubMed1989

Inducibility of liver cytosolic aldehyde dehydrogenase activity in various animal species.

1. The inducibility of hepatic cytosolic aldehyde dehydrogenase activity was studied in rat, mouse, guinea pig, chicken, frog, salamander and rainbow trout, by using two different types of inducers of drug metabolism. 2. Phenobarbital (a type I inducer of drug metabolizing enzymes) increased total liver cytosolic aldehyde dehydrogenase activity (up to 20-fold) in a genetically defined substrain of responsive rats (RR) and only slightly, if at all, in a non-responsive substrain (rr). On the contrary, both types of rats showed a highly induced aldehyde dehydrogenase activity after treatment with methylcholanthrene (a type II inducer). Phenobarbital is affecting mainly an isozyme of aldehyde dehydrogenase which is best measured with propionaldehyde as the substrate and NAD as the coenzyme (P/NAD). 3. Administration of phenobarbital to mice produced only a slight increase (2-fold) in the P/NAD aldehyde dehydrogenase activity. 4. Methylcholanthrene treatment caused a 2-fold increase of the hepatic P/NAD aldehyde dehydrogenase activity in the chicken. 5. In the guinea pig, phenobarbital produced an approximate 3-fold increase of the P/NAD activity. Methylcholanthrene had a similar effect, although to a lesser extent. 6. In the salamander, a 4-fold increase was detected in the enzyme activity measured with benzaldehyde as the substrate and NADP as the coenzyme (B/NADP), after treatment with either phenobarbital or methylcholanthrene. 7. The hepatic aldehyde dehydrogenase activities were found unchanged in the rainbow trout, after treatment with phenobarbital or 2,3,7,8-tetrachlorodibenzo-p-dioxin. 8. The rat model remains the only one examined that shares with human hepatocytes strong inducibility of the B/NADP aldehyde dehydrogenase isozyme upon treatment with polycyclic aromatic hydrocarbons.

Aldehyde Dehydrogenase↗
PubMed2005

Neuroanatomical distribution of vasotocin and mesotocin in two urodele amphibians (Plethodon shermani and Taricha granulosa) based on in situ hybridization histochemistry.

Previous research suggests that considerable species-specific variation exists in the neuroanatomical distributions of arginine vasotocin (AVT) and mesotocin (MST), non-mammalian homologues of vasopressin and oxytocin. An earlier study in rough-skinned newts (Taricha granulosa) indicated that the neuroanatomical distribution of cells labeled for AVT-immunoreactivity (ir) was greater in this urodele amphibian than in any other species. It was unknown whether the widespread distribution of AVT-ir is unique to T. granulosa or a feature common among salamanders. Using in situ hybridization (ISH) histochemistry and gene-specific riboprobes, the current study labeled AVT and MST mRNA in T. granulosa and the red-legged salamander (Plethodon shermani). In T. granulosa, AVT ISH-labeled cells were found to be widespread and localized in brain areas including the dorsal and medial pallium, lateral and medial septum, bed nucleus of the stria terminalis, amygdala, preoptic area, ventral hypothalamus, nucleus isthmus, tectum mesencephali, inferior colliculus, and hindbrain. In P. shermani, the distribution of AVT ISH-labeled neurons matched that of T. granulosa, except in the lateral septum, ventral hypothalamus, and inferior colliculus, but did however include labeled cell bodies in the lateral pallium. The distribution of MST ISH-labeled cells was more restricted than AVT ISH labeling and was limited to regions of the preoptic area and ventral thalamus, which is consistent with the limited distribution of MST/OXY in other vertebrates. These findings support the conclusion that urodele amphibians possess a well-developed vasotocin system, perhaps more extensive than other vertebrate taxa.

Animals↗
PubMed2006

Palmitylation of cone opsins.

Palmitylation is a widespread modification in G-protein-coupled receptors and often a dynamic process. In rhodopsins, palmitylation is static on C322/C323. Red/green (M/LWS) cone opsins have no cysteines at corresponding positions and no palmitylation. Blue (SWS2) cone opsins have a single corresponding cysteine and mass spectrometric analysis showed partial palmitylation of salamander SWS2 cone opsin. Ultraviolet (SWS1) cone opsins have one corresponding cysteine, but only unpalmitylated opsin was observed for mouse and salamander. The results show that the static palmitylation found on rhodopsin is not found on cone opsins and suggest the possibility of an unidentified role for opsin palmitylation in cones.

Animals↗
PubMed2006

In situ localization of plethodontid courtship pheromone mRNA in formalin-fixed tissue.

Male plethodontid salamanders produce courtship pheromones that increase female receptivity. Three protein components of the courtship pheromone cocktail have been characterized in the mental gland of Plethodon shermani, the red-legged salamander: plethodontid receptivity factor (PRF), plethodontid modulating factor (PMF), and sodefrin precursor-like factor (SPF). In this study, a streamlined in situ hybridization (ISH) protocol, employing a biotinylated oligonucleotide probe, is used to visualize the sites of pheromone expression in formalin-fixed paraffin-embedded P. shermani mental gland and post-cloacal tail tissue. Results corroborate previous RT-PCR studies on pheromone expression. PRF and PMF are highly expressed in P. shermani mental gland, while SPF expression is more variable. None of the tested pheromones is expressed in dorsal or ventral tail glands. The reported protocol is simple, rapid, and effective, allowing visualization of high-copy mRNA transcript in formalin-fixed tissue.

Animals↗
PubMed1995

Cyclic GMP diffusion coefficient in rod photoreceptor outer segments.

Cyclic GMP (cGMP) is the intracellular messenger that mediates phototransduction in retinal rods. As photoisomerizations of rhodopsin molecules are local events, the longitudinal diffusion of cGMP in the rod outer segment should be a contributing factor to the response of the cell to light. We have employed the truncated rod outer segment preparation from bullfrog (Rana catesbeiana) and tiger salamander (Ambystoma tigrinum) to measure the cGMP diffusion coefficient. In this preparation, the distal portion of a rod outer segment was drawn into a suction pipette for measuring membrane current, and the rest of the cell was then sheared off with a glass probe, allowing bath cGMP to diffuse into the outer segment and activate the cGMP-gated channels on the surface membrane. Addition and removal of bath cGMP were fast enough to produce effectively step changes in cGMP concentration at the open end of the outer segment. When cGMP hydrolysis is inhibited by isobutylmethylxanthine (IBMX), the equation for the diffusion of cGMP inside the truncated rod outer segment has a simple analytical solution, which we have used to analyze the rise and decay kinetics of the cGMP-elicited currents. From these measurements we have obtained a cGMP diffusion coefficient of approximately 70 x 10(-8) cm2 s-1 for bullfrog rods and approximately 60 x 10(-8) cm2 s-1 for tiger salamander rods. These values are six to seven times lower than the expected value in aqueous solution. The estimated diffusion coefficient is the same at high (20-1000 microM) and low (5-10 microM) concentrations of cGMP, suggesting no significant effect from buffering over these concentration ranges.

1-Methyl-3-isobutylxanthine↗
PubMed1978

Electron microscopic localization of Mg2+ -dependent adenosine triphosphatase activity in the amphibian pancreas (Salamandra salamandra L. and Rana esculenta L.).

The ultrastructural distribution of Mg2+ -dependent adenosine triphosphatase (ATPase) activity has been investigated in the salamander and frog pancreas by using glutaraldehyde fixations and a modified Wachstein-Meisel reaction medium. In both species the reaction product (lead phosphate) was found associated with the plasma membrane external side of all islet cell types (B-, A- and D-cells) and of acinar and ductular/centro-acinar cells. Except the apical pole of salamander acinar and centro-acinar cells, usually devoid of reaction, no preferential distribution of enzyme activity depending on endocrine or exocrine cell aspects could be observed. Other specific enzyme localizations included the mitochondria matrices, nucleoles, condensed nuclear chromatin, periaxolemmal spaces in nerve bundles and sometimes the cleft of neuro-glandular junctions. The occurrence of reaction deposits in connective tissue, in the cytoplasm of both islet and exocrine cells and in the nerve fiber axoplasm was considered as a possible diffusion artifact. The reaction intensity, but not its distribution, varied sensibly with the incubation period. 2-iodoacetamide and p-chlormercuribenzoic acid decreased the amount of reaction deposits at the level of all reactive sites and especially in mitochondria. The specificity of Mg2+ -ATPase demonstration in this paper is analysed taking into account several inherent shortcomings of the Wachstein-Meisel incubation medium and of the fixative. The different enzyme localizations, as well as their functional significances are discussed in relation with the findings of other authors.

Adenosine Triphosphatases↗
PubMed1978

Metabolism of benzo(a)pyrene in animals with high aryl hydrocarbon hydroxylase levels and high rates of spontaneous cancer.

Ambystoma tigrinum found in a sewage polluted pond had high levels of aryl hydrocarbon hydroxylase (AHH) activity that decreased to the basal level of control animals after being held several months in clean water. The qualitative formation of benzo(a)pyrene (BP) metabolites by salamander hepatic microsomes was similar to those seen for other species. Inhibition of epoxide hydrase activity did not alter the total metabolite production but did change the ratio of metabolites. A correlation appears to exist between high AHH induction, the presence of polycyclic hydrocarbon pollutants, and the high rate of spontaneous cancer in salamanders.

Ambystoma↗
PubMed2000

The role of mating preferences in shaping interspecific divergence in mating signals in vertebrates.

Vertebrates represent one of the best-studied groups in terms of the role that mating preferences have played in the evolution of exaggerated secondary sexual characters and mating behaviours within species. Vertebrate species however, also exhibit enormous interspecific diversity in features of mating signals that has potentially led to reproductive isolation and speciation in many groups. The role that sexual selection has played in interspecific divergence in mating signals has been less fully explored. This review summarizes our current knowledge of how mating preferences within species have shaped interspecific divergence in mate recognition signals among the major vertebrate groups. Certain signal modalities appear to characterize mating signal diversification among different vertebrate taxa. Acoustic signals play an important role in mating decisions in anuran amphibians and birds. Here, different properties of the signal may convey information regarding individual, neighbor and species recognition. Mating preferences for particular features of the acoustic signal have led to interspecific divergence in calls and songs. Divergence in morphological traits such as colouration or ornamentation appears to be important in interspecific diversity in certain groups of fishes and birds. Pheromonal signals serve as the primary basis for species-specific mating cues in many salamander species, most mammals and even some fishes. The evolution of interspecific divergence in elaborate courtship displays may have played an important role in speciation of lizards, and particular groups of fishes, salamanders, birds and mammals. While much research has focused on the importance of mating preferences in shaping the evolution of these types of mating signals within species, the link between intraspecific preferences and interspecific divergence and speciation remains to be more fully tested. Future studies should focus on identifying how variation in mating preferences within a species shapes interspecific diversity in features of mating signals in order to better understand how sexual selection may have led to speciation in vertebrates.

Journal Article↗
PubMed2002

Inducing cellular dedifferentiation: a potential method for enhancing endogenous regeneration in mammals.

Salamanders have the remarkable ability to regenerate lost body parts and injured organs. This regenerative ability requires fully-differentiated cells in the vicinity of the injury to dedifferentiate, proliferate, and then redifferentiate to form the specialized cells that comprise the regenerated structure or organ. The dedifferentiation stage plays a crucial role in the regenerative response and distinguishes the salamander from other vertebrates with more limited regenerative abilities. Recently, several investigators have shown that certain mammalian cell types can be induced to dedifferentiate to progenitor cells when stimulated with the appropriate signals. This discovery opens the possibility that researchers might one day enhance the endogenous regenerative capacity of mammals by inducing cellular dedifferentiation in vivo.

Animals↗
PubMed2000

Amphibian respiratory diseases.

This article describes the diseases that commonly affect the lungs, gills, and nasal passages of frogs, toads, and salamanders. Although these organs often are affected by systemic infections, only diseases that are limited to the respiratory system are discussed. Gills of aquatic amphibians commonly are affected by poor water quality, infestations by protozoa or trematodes, or infections by water molds. Nematodes in the genus Rhabias are significant pulmonary pathogens in frogs and toads. The nasal passages of frogs, toads, and salamanders may be parasitized by Bufolucilia fly larvae.

Amphibians↗
PubMed2002

Peptide and protein pheromones in amphibians.

Purification, characterization and biological activity of urodele and anuran sex-pheromones were reviewed. Female-attracting pheromones obtained from the abdominal gland of Cynops pyrrhogaster and C. ensicauda males are peptides consisting of 10 amino acid residues being designated sodefrin and silefrin, respectively. Each pheromone attracted only conspecific females. Molecular cloning of cDNAs encoding sodefrin and silefrin revealed that both are generated from precursor proteins. Synthesis of these pheromones is regulated by prolactin (PRL) and androgen. Responsiveness of the female vomeronasal epithelium to sodefrin is enhanced by PRL and estrogen. The submandibular gland of the male terrestrial salamander, Plethodon jardani secretes a 22-kD proteinaceous pheromone that enhances female receptivity. It was revealed that every salamander synthesizes multiple isoforms of this pheromone, Plethodontid receptivity factor. The magnificent tree frog, Litoria splendida breed in an aquatic environment. The skin glands of the male secrete a female-attracting peptide pheromone, splendipherin, comprising 25 amino acid residues. The significance of the structure of the amphibian sex-pheromone as peptide and protein is discussed in terms of their species specificity.

Amphibians↗
PubMedDate not supplied

Evidence for the prolonged photoactivated lifetime of an analogue visual pigment containing 11-cis 9-desmethylretinal.

Following bright flashes, rod photoreceptors exhibit a period of photocurrent saturation that increases linearly with the logarithm of flash intensity. In a recent report, Pepperberg et al. (1992) presented evidence that the slope of the function relating the saturation period (T) to the natural logarithm of flash intensity (ln If) represents the exponential lifetime (tau) of photoactivated visual pigment: tau = delta T/delta [ln If]. In salamander rods, 11-cis 9-desmethylretinal combines with opsin to form 9-desmethyl rhodopsin. Dim flash responses mediated by this analogue visual pigment exhibited slow recovery kinetics relative to those of native pigment (Corson et al., 1991). This observation raises the hypothesis that the physiological lifetime of photoactivated 9-desmethyl rhodopsin is substantially longer than that of native visual pigment. To test this hypothesis, we have examined the relation between the period of photocurrent saturation and flash intensity in salamander rods containing a mixture of the two pigments. Brief stimuli at two widely separated wavelengths (440 and 640 nm) elicited saturating photocurrent responses that were preferentially mediated by 9-desmethyl rhodopsin or residual native pigment, respectively. Plots of T vs. ln If revealed a linear increase in the period of response saturation over a large range of saturating intensities at both wavelengths. However, the slope of the relation between T and ln If with 440-nm flashes was more than twice as large (4.1 +/- 0.5 s, n = 5) as that measured with 640-nm flashes (1.7 +/- 0.4 s). For rods subjected only to bleaching of the native pigment, or to bleaching and resensitization with 11-cis retinal, the slope of the relation between T and ln If remained independent of wavelength and indistinguishable from that of native pigment in unbleached cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2001

Calcium-activated potassium current clamps the dark potential of vertebrate rods.

Vertebrate photoreceptors respond to light with a graded hyperpolarization from a membrane potential in the dark of approximately -35 mV. The present work investigates the physiological role of the Ca2+-activated K+ current in the photovoltage generation in mechanically isolated rods from salamander retina. Membrane current or voltage in isolated rods was recorded from light- and dark-adapted rods under voltage- or current-clamp conditions, respectively. In light-adapted rods of the salamander, selective blockade of Ca2+-activated K+ channels by means of charybdotoxin depolarized the plasma membrane of current-clamped rods by approximately 30 mV, from a resting potential of approximately -35 mV. A similar depolarization was observed if external Ca2+ (1 mM) was substituted with Ba2+ or Sr2+. Under control conditions, the injection of currents of increasing amplitude (up to -100 pA, to mimic the current entering the rod outer segment) could not depolarize the membrane potential beyond a saturating value of approximately -20 mV. However, in the presence of charybdotoxin, rods depolarized up to +20 mV. In experiments with dark-adapted current-clamped rods, charybdotoxin perfusion lead to transient depolarizations up to 0 mV and steady-state depolarizations of approximately 5 mV above the dark resting potential. Finally, the recovery phase of the voltage response to a flash of light in the presence of charybdotoxin showed a transient overshoot of the membrane potential. It was concluded that Ca2+-activated K+ current is necessary for clamping the rod photovoltage to values close to the dark potential, thus allowing faithful single photon detection and correct synaptic transmission.

Ambystoma mexicanum↗
PubMedDate not supplied

An ependymal cell culture system for the study of spinal cord regeneration.

Successful regeneration of lesioned adult spinal cord in urodele (caudate) amphibians requires the action of injury-responsive ependymal cells (ependymoglia). The epithelial-to-mesenchymal transformation of ependymal cells following transection of the salamander spinal cord and the subsequent reformation of an epithelial tube have been described previously. A complete tissue culture model system has now been devised to study mesenchymal ependymal cells, epithelial ependymal cells, and ependymal/neuronal interactions in vitro. Here, we review critical aspects of substrate and growth factor environments required to produce mesenchymal ependymal cells in culture and present the first culture system for epithelial salamander ependymal cells and central nervous system neurons suitable for cell-cell interaction studies. Critical to ependymal epithelialization in culture is the removal of epidermal growth factor and addition of thrombin. Epithelialization occurs on tissue culture plastic as well as on permeable culture substrates. This culture system can now be used to determine the initial trigger for the ependymal response. A preliminary examination of ependymal/neuronal interactions shows that coculture of mesenchymal ependymal cells and central nervous system neurons prolongs survival of the neurons.

Animals↗
PubMed2007

Global patterns of diversification in the history of modern amphibians.

The fossil record of modern amphibians (frogs, salamanders, and caecilians) provides no evidence for major extinction or radiation episodes throughout most of the Mesozoic and early Tertiary. However, long-term gradual diversification is difficult to reconcile with the sensitivity of present-day amphibian faunas to rapid ecological changes and the incidence of similar environmental perturbations in the past that have been associated with high turnover rates in other land vertebrates. To provide a comprehensive overview of the history of amphibian diversification, we constructed a phylogenetic timetree based on a multigene data set of 3.75 kb for 171 species. Our analyses reveal several episodes of accelerated amphibian diversification, which do not fit models of gradual lineage accumulation. Global turning points in the phylogenetic and ecological diversification occurred after the end-Permian mass extinction and in the late Cretaceous. Fluctuations in amphibian diversification show strong temporal correlation with turnover rates in amniotes and the rise of angiosperm-dominated forests. Approximately 86% of modern frog species and >81% of salamander species descended from only five ancestral lineages that produced major radiations in the late Cretaceous and early Tertiary. This proportionally late accumulation of extant lineage diversity contrasts with the long evolutionary history of amphibians but is in line with the Tertiary increase in fossil abundance toward the present.

Amphibians↗
PubMed2003

D-serine and serine racemase are present in the vertebrate retina and contribute to the physiological activation of NMDA receptors.

d-serine has been proposed as an endogenous modulator of N-methyl-d-aspartate (NMDA) receptors in many brain regions, but its presence and function in the vertebrate retina have not been characterized. We have detected d-serine and its synthesizing enzyme, serine racemase, in the retinas of several vertebrate species, including salamanders, rats, and mice and have localized both constituents to Müller cells and astrocytes, the two major glial cell types in the retina. Physiological studies in rats and salamanders demonstrated that, in retinal ganglion cells, d-serine can enhance excitatory currents elicited by the application of NMDA, as well as the NMDA receptor component of light-evoked synaptic responses. Application of d-amino acid oxidase, which degrades d-serine, reduced the magnitude of NMDA receptor-mediated currents, raising the possibility that endogenous d-serine serves as a ligand for setting the sensitivity of NMDA receptors under physiological conditions. These observations raise exciting new questions about the role of glial cells in regulating the excitability of neurons through release of d-serine.

Animals↗