African Clawed Frog Xenopus Laevis Research Articles

Altered thyroid hormone levels affect body condition at metamorphosis in larvae of Xenopus laevis.

Chemical, physical and biological environmental stressors may affect the endocrine system, such as the thyroid hormone (TH) axis in larval amphibians with consequences for energy partitioning among development, growth and metabolism. We studied the effects of two TH level affecting compounds, exogenous l-thyroxine (T4 ) and sodium perchlorate (SP), on various measures of development and body condition in larvae of the African clawed frog (Xenopus laevis). We calculated the scaled mass index, hepatosomatic index and relative tail muscle mass as body condition indices to estimate fitness. Altered TH levels significantly altered the growth, development, survival and body condition in metamorphic larvae in different directions. While exogeno us T4 reduced growth and accelerated development, SP treatment increased growth but slowed down development. Altered TH levels improved body conditions in both treatments and particularly in larvae of the SP treatment but to the detriment of lower survival rates in both TH level altering treatments. The hepatosomatic index was negatively affected by exogenous T4 , but not by SP treatment indicating a lower lipid reserve in the liver in larvae of T4 treatment. These altered TH levels as caused by several environmental stressors may have an influence on individual fitness across life, as body condition at the onset of metamorphosis determines metamorphic and juvenile survival. Further research is needed to determine synergetic effects of environmental stressors on TH levels and its effects on physiological traits such as metabolic rate.

The TMEM16A channel mediates the fast polyspermy block in Xenopus laevis.

In externally fertilizing animals, such as sea urchins and frogs, prolonged depolarization of the egg immediately after fertilization inhibits the entry of additional sperm-a phenomenon known as the fast block to polyspermy. In the African clawed frog Xenopus laevis, this depolarization is driven by Ca2+-activated Cl- efflux. Although the prominent Ca2+-activated Cl- currents generated in immature X. laevis oocytes are mediated by X. laevis transmembrane protein 16a (xTMEM16A) channels, little is known about the channels that contribute to the fast block in mature eggs. Moreover, the gamete undergoes a gross transformation as it develops from an immature oocyte into a fertilization-competent egg. Here, we report the results of our approach to identify the Ca2+-activated Cl- channel that triggers the fast block. By querying published proteomic and RNA-sequencing data, we identify two Ca2+-activated Cl- channels expressed in fertilization-competent X. laevis eggs: xTMEM16A and X. laevis bestrophin 2A (xBEST2A). By exogenously expressing xTMEM16A and xBEST2A in axolotl cells lacking endogenous Ca2+-activated currents, we characterize the effect of inhibitors on currents mediated by these channels. None of the inhibitors tested block xBEST2A currents specifically. However, 2-(4-chloro-2-methylphenoxy)-N-[(2-methoxyphenyl)methylideneamino]-acetamide (Ani9) and N-((4-methoxy)-2-naphthyl)-5-nitroanthranilic acid (MONNA) each reduce xTMEM16A currents by more than 70% while only nominally inhibiting those generated by xBEST2A. Using whole-cell recordings during fertilization, we find that Ani9 and MONNA effectively diminish fertilization-evoked depolarizations. Additionally, these inhibitors lead to increased polyspermy in X. laevis embryos. These results indicate that fertilization activates TMEM16A channels in X. laevis eggs and induces the earliest known event triggered by fertilization: the fast block to polyspermy.

Multiple massive domestication and recent amplification of Kolobok superfamily transposons in the clawed frog Xenopus

BackgroundDNA transposons are generally destroyed by mutations and have short lifespans in hosts, as they are neutral or harmful to the host and therefore not conserved by natural selection. The clawed frog Xenopus harbors many DNA transposons and certain families, such as T2-MITE, have extremely long lives. These have ancient origins, but have shown recent transposition activity. In addition, certain transposase genes may have been “domesticated” by Xenopus and conserved over long time periods by natural selection. The aim of this study was to elucidate the evolutionary interactions between the host and the long-lived DNA transposon family it contains. Here, we investigated the molecular evolution of the Kolobok DNA transposon superfamily. Kolobok is thought to contribute to T2-MITE transposition.ResultsIn the diploid western clawed frog Xenopus tropicalis and the allotetraploid African clawed frog Xenopus laevis, we searched for transposase genes homologous to those in the Kolobok superfamily. To determine the amplification and domestication of these genes, we used molecular phylogenetics and analyses of copy numbers, conserved motifs, orthologous gene synteny, and coding sequence divergence between the orthologs of X. laevis and X. tropicalis, or between those of two distant X. tropicalis lineages. Among 38 X. tropicalis and 24 X. laevis prospective transposase genes, 10 or more in X. tropicalis and 14 or more in X. laevis were apparently domesticated. These genes may have undergone multiple independent domestications from before the divergence of X. laevis and X. tropicalis. In contrast, certain other transposases may have retained catalytic activity required for transposition and could therefore have been recently amplified.ConclusionMultiple domestication of certain transposases and prolonged conservation of the catalytic activity in others suggest that Kolobok superfamily transposons were involved in complex, mutually beneficial relationships with their Xenopus hosts. Some transposases may serve to activate long-lived T2-MITE subfamilies.

Spindle assembly in egg extracts of the Marsabit clawed frog, Xenopus borealis.

Egg extracts of the African clawed frog Xenopus laevis have provided a cell-free system instrumental in elucidating events of the cell cycle, including mechanisms of spindle assembly. Comparison with extracts from the diploid Western clawed frog, Xenopus tropicalis, which is smaller at the organism, cellular and subcellular levels, has enabled the identification of spindle size scaling factors. We set out to characterize the Marsabit clawed frog, Xenopus borealis, which is intermediate in size between the two species, but more recently diverged in evolution from X. laevis than X. tropicalis. X. borealis eggs were slightly smaller than those of X. laevis, and slightly smaller spindles were assembled in egg extracts. Interestingly, microtubule distribution across the length of the X. borealis spindles differed from both X. laevis and X. tropicalis. Extract mixing experiments revealed common scaling phenomena among Xenopus species, while characterization of spindle factors katanin, TPX2, and Ran indicate that X. borealis spindles possess both X. laevis and X. tropicalis features. Thus, X. borealis egg extract provides a third in vitro system to investigate interspecies scaling and spindle morphometric variation.

Effects of environmentally relevant sub-chronic atrazine concentrations on African clawed frog (Xenopus laevis) survival, growth and male gonad development

Sub-chronic toxicity of environmentally relevant atrazine concentrations on exposed tadpoles and adult male African clawed frogs (Xenopus laevis) was evaluated in a quality controlled laboratory for 90 days. The aim of this study was to determine the effects of atrazine on the survival, growth and gonad development of African clawed frogs. After exposure of tadpoles to atrazine concentrations of 0 (control), 0.01, 200 and 500 μg L−1 in water, mortality rates of 0, 0, 3.3 and 70% respectively were recorded for the 90 day exposure period. Morphometry showed significantly reduced tadpole mass in the 500 μg L−1 atrazine exposed tadpoles (p < 0.05). Light microscopy on testes of adult frogs exposed to the same atrazine concentrations using hematoxylin and eosin (H&E) and Van Gieson staining techniques revealed gonadal atrophy, disruption of germ cell lines, seminiferous tubule structure damage and formation of extensive connective tissue around seminiferous tubules of frogs exposed to 200 μg L−1 and 500 μg L−1 atrazine concentrations. Ultrastructural analysis of the cellular organelles using transmission electron microscopy (TEM) revealed significant amounts of damaged mitochondria in testosterone producing Leydig cells as well as Sertoli cells. Biochemical analysis revealed reduced serum testosterone levels in adult frogs at all exposure levels as well as presence of six atrazine metabolites in frog serum and liver. The results indicate that atrazine concentrations greater than the calculated LC50 of 343.7 μg L−1 cause significant mortality in tadpoles, while concentrations ≥200 μg L−1 adversely affect reproductive health of adult frogs and development of tadpoles sub-chronically exposed to atrazine.

Embryonic lethality is not sufficient to explain hourglass-like conservation of vertebrate embryos

BackgroundUnderstanding the general trends in developmental changes during animal evolution, which are often associated with morphological diversification, has long been a central issue in evolutionary developmental biology. Recent comparative transcriptomic studies revealed that gene expression profiles of mid-embryonic period tend to be more evolutionarily conserved than those in earlier or later periods. While the hourglass-like divergence of developmental processes has been demonstrated in a variety of animal groups such as vertebrates, arthropods, and nematodes, the exact mechanism leading to this mid-embryonic conservation remains to be clarified. One possibility is that the mid-embryonic period (pharyngula period in vertebrates) is highly prone to embryonic lethality, and the resulting negative selections lead to evolutionary conservation of this phase. Here, we tested this “mid-embryonic lethality hypothesis” by measuring the rate of lethal phenotypes of three different species of vertebrate embryos subjected to two kinds of perturbations: transient perturbations and genetic mutations.ResultsBy subjecting zebrafish (Danio rerio), African clawed frog (Xenopus laevis), and chicken (Gallus gallus) embryos to transient perturbations, namely heat shock and inhibitor treatments during three developmental periods [early (represented by blastula and gastrula), pharyngula, and late], we found that the early stages showed the highest rate of lethal phenotypes in all three species. This result was corroborated by perturbation with genetic mutations. By tracking the survival rate of wild-type embryos and embryos with genetic mutations induced by UV irradiation in zebrafish and African clawed frogs, we found that the highest decrease in survival rate was at the early stages particularly around gastrulation in both these species.ConclusionIn opposition to the “mid-embryonic lethality hypothesis,” our results consistently showed that the stage with the highest lethality was not around the conserved pharyngula period, but rather around the early period in all the vertebrate species tested. These results suggest that negative selection by embryonic lethality could not explain hourglass-like conservation of animal embryos. This highlights the potential contribution of alternative mechanisms such as the diversifying effect of positive selections against earlier and later stages, and developmental constraints which lead to conservation of mid-embryonic stages.

The right thalamus may play an important role in anesthesia-awakening regulation in frogs.

BackgroundPrevious studies have shown that the mammalian thalamus is a key structure for anesthesia-induced unconsciousness and anesthesia-awakening regulation. However, both the dynamic characteristics and probable lateralization of thalamic functioning during anesthesia-awakening regulation are not fully understood, and little is known of the evolutionary basis of the role of the thalamus in anesthesia-awakening regulation.MethodsAn amphibian species, the South African clawed frog (Xenopus laevis) was used in the present study. The frogs were immersed in triciane methanesulfonate (MS-222) for general anesthesia. Electroencephalogram (EEG) signals were recorded continuously from both sides of the telencephalon, diencephalon (thalamus) and mesencephalon during the pre-anesthesia stage, administration stage, recovery stage and post-anesthesia stage. EEG data was analyzed including calculation of approximate entropy (ApEn) and permutation entropy (PE).ResultsBoth ApEn and PE values differed significantly between anesthesia stages, with the highest values occurring during the awakening period and the lowest values during the anesthesia period. There was a significant correlation between the stage durations and ApEn or PE values during anesthesia-awakening cycle primarily for the right diencephalon (right thalamus). ApEn and PE values for females were significantly higher than those for males.DiscussionApEn and PE measurements are suitable for estimating depth of anesthesia and complexity of amphibian brain activity. The right thalamus appears physiologically positioned to play an important role in anesthesia-awakening regulation in frogs indicating an early evolutionary origin of the role of the thalamus in arousal and consciousness in land vertebrates. Sex differences exist in the neural regulation of general anesthesia in frogs.

Invasive African clawed frogs in California: A reservoir for or predator against the chytrid fungus?

Amphibian species are experiencing population declines due to infection by the fungal pathogen, Batrachochytrium dendrobatidis (Bd). The African clawed frog (Xenopus laevis), an asymptomatic carrier of Bd, has been implicated in the spread of this pathogen through global trade and established invasive populations on several continents. However, research has not explored the relationships of both life stages of this amphibian with Bd. While the post-metamorphic individuals may act as a reservoir, spreading the infection to susceptible species, the filter-feeding larvae may consume the motile Bd zoospores from the water column, potentially reducing pathogen abundance and thus the likelihood of infection. We explore these contrasting processes by assessing Bd prevalence and infection intensities in field populations of post-metamorphic individuals, and performing laboratory experiments to determine if larval X. laevis preyed upon Bd zoospores. The water flea, Daphnia magna, was included in the Bd consumption trials to compare consumption rates and to explore whether intraguild predation between the larval X. laevis and Daphnia may occur, potentially interfering with control of Bd zoospores by Daphnia. Field surveys of three X. laevis populations in southern California, in which 70 post-metamorphic individuals were tested for Bd, found 10% infection prevalence. All infected individuals had very low infection loads (all Bd loads were below 5 zoospore equivalents). Laboratory experiments found that larval X. laevis consume Bd zoospores and therefore may reduce Bd abundance and transmission between amphibians. However, metamorphic and juvenile X. laevis exhibited intraguild predation by consuming Daphnia, which also prey upon Bd zoospores. The results suggest that X laevis is not a large reservoir for Bd and its larval stage may offer some reduction of Bd transmission through direct predation.

Thyroid hormone levels and temperature during development alter thermal tolerance and energetics of Xenopus laevis larvae.

Environmental variation induced by natural and anthropogenic processes including climate change may threaten species by causing environmental stress. Anuran larvae experiencing environmental stress may display altered thyroid hormone (TH) status with potential implications for physiological traits. Therefore, any capacity to adapt to environmental changes through plastic responses provides a key to determining species vulnerability to environmental variation. We investigated whether developmental temperature (T dev), altered TH levels and whether the interactive effect of both affect standard metabolic rate (SMR), body condition (BC), survival and thermal tolerance in larvae of the African clawed frog (Xenopus laevis) reared at five temperatures with experimentally altered TH levels. At metamorphosis, SMR, BC and survival were significantly affected by T dev, TH status and their interaction with the latter often intensified impacts. Larvae developing at warmer temperatures exhibited significantly higher SMRs and BC was reduced at warm T dev and high TH levels suggesting decreased ability to acclimate to variation in temperature. Accordingly, tadpoles that developed at warm temperatures had higher maximum thermal limits but more narrow thermal tolerance windows. High and low TH levels decreased and increased upper thermal limits, respectively. Thus, when experiencing both warmer temperatures and environmental stress, larvae may be less able to compensate for changes in T dev. Our results demonstrate that physiological traits in larvae of X. laevis are strongly affected by increased TH levels and warmer temperatures. Altered TH levels and increasing T dev due to global change may result in a reduced capacity for physiological plasticity. This has far reaching consequences since the energetic requirement at the onset of metamorphosis is known to determine metamorphic success and thus, is indirectly linked to individual fitness in later life stages.

Influence of temperature on reproduction and length of metamorphosis in Xenopus laevis (Amphibia: Anura)

Research was conducted on breeding and metamorphosis of a species of the African clawed frog (Xenopus laevis). The experimental group consisted of 10 individuals each of albino and natural forms. Specimens were divided into pairs and numbered for better tracking. In experimental ponds, conditions simulating the mating season were created, which led to 100% reproduction of all the pairs. The aim of the work was to determine the effects of temperature on the gender of resulting metamorphic specimens, and how the length of metamorphosis affects their gender. Specimens were divided into two groups. The first group consisted of specimens bred at 19°C, and the second group of specimens were bred at 24°C. Mating covered the period from 5 May to 25 June 2016. Observations were completed on 31 October 2016. Five hundred and fourteen individual metamorphic specimens were bred during the experiment. The research shows that the length of metamorphosis has an effect on gender, in favour of females. The effect of temperature on gender was not confirmed. The experimental results can be used as a basis for similar research, since this topic has not been discussed previously.

Evaluation of toxicity of waters from the surface sources of water supply and drinking water of Zhytomyr city by the method of short-term chronic biotesting

The goal of the researches is to increase the informativeness of testing water from the surface sources of water supply and drinking water at the expense of the use of african clawed frog. Method of the researches is biotesting with determination of test sign of immobilization the marsh frog (Rana ridibunda Pallas, 1771) and african clawed frog (Xenopus laevis Daudin, 1802) at term 8-day exposition of individuals. During a biotesting for lack of immobilization R. ridibunda it is not discovered toxicity of water, but on the basis of immobilization X. laevis the low level of toxicity of them in within 25–50% is defined. Assessment of chronic toxicity of waters from the surface sources of water supply and drinking water is expedient to conduct, using the biotesting with X. laevis. It is shown the high sensibility of these test-organisms in comparison with R. ridibunda (in average to 28.2% in acute lethal tests with exposition time 4 day and to 41.3% in short-term chronic tests with exposition time 8 day).

Microprobe Capillary Electrophoresis Mass Spectrometry for Single-cell Metabolomics in Live Frog (Xenopus laevis) Embryos.

The quantification of small molecules in single cells raises new potentials for better understanding the basic processes that underlie embryonic development. To enable single-cell investigations directly in live embryos, new analytical approaches are needed, particularly those that are sensitive, selective, quantitative, robust, and scalable to different cell sizes. Here, we present a protocol that enables the in situ analysis of metabolism in single cells in freely developing embryos of the South African clawed frog (Xenopus laevis), a powerful model in cell and developmental biology. This approach uses a capillary microprobe to aspirate a defined portion from single identified cells in the embryo, leaving neighboring cells intact for subsequent analysis. The collected cell content is analyzed by a microscale capillary electrophoresis electrospray ionization (CE-ESI) interface coupled to a high-resolution tandem mass spectrometer. This approach is scalable to various cell sizes and compatible with the complex three-dimensional structure of the developing embryo. As an example, we demonstrate that microprobe single-cell CE-ESI-MS enables the elucidation of metabolic cell heterogeneity that unfolds as a progenitor cell gives rise to descendants during development of the embryo. Besides cell and developmental biology, the single-cell analysis protocols described here are amenable to other cell sizes, cell types, or animal models.

Microprobe Capillary Electrophoresis Mass Spectrometry for Single-cell Metabolomics in Live Frog (&lt;em&gt;Xenopus laevis&lt;/em&gt;) Embryos

The quantification of small molecules in single cells raises new potentials for better understanding the basic processes that underlie embryonic development. To enable single-cell investigations directly in live embryos, new analytical approaches are needed, particularly those that are sensitive, selective, quantitative, robust, and scalable to different cell sizes. Here, we present a protocol that enables the in situ analysis of metabolism in single cells in freely developing embryos of the South African clawed frog (Xenopus laevis), a powerful model in cell and developmental biology. This approach uses a capillary microprobe to aspirate a defined portion from single identified cells in the embryo, leaving neighboring cells intact for subsequent analysis. The collected cell content is analyzed by a microscale capillary electrophoresis electrospray ionization (CE-ESI) interface coupled to a high-resolution tandem mass spectrometer. This approach is scalable to various cell sizes and compatible with the complex three-dimensional structure of the developing embryo. As an example, we demonstrate that microprobe single-cell CE-ESI-MS enables the elucidation of metabolic cell heterogeneity that unfolds as a progenitor cell gives rise to descendants during development of the embryo. Besides cell and developmental biology, the single-cell analysis protocols described here are amenable to other cell sizes, cell types, or animal models.

Xenopus laevis as a Model Organism for the Study of Spinal Cord Formation, Development, Function and Regeneration.

The spinal cord is the first central nervous system structure to develop during vertebrate embryogenesis, underscoring its importance to the organism. Because of its early formation, accessibility to the developing spinal cord in amniotes is challenging, often invasive and the experimental approaches amenable to model systems like mammals are limited. In contrast, amphibians, in general and the African-clawed frog Xenopus laevis, in particular, offer model systems in which the formation of the spinal cord, the differentiation of spinal neurons and glia and the establishment of spinal neuron and neuromuscular synapses can be easily investigated with minimal perturbations to the whole organism. The significant advances on gene editing and microscopy along with the recent completion of the Xenopus laevis genome sequencing have reinvigorated the use of this classic model species to elucidate the mechanisms of spinal cord formation, development, function and regeneration.

Xenopus larvae swivel reflex depends on contrast

Some photographers take advantage of the blurring effect produced when animals and humans move past the lens at high speed to produce extraordinary images, but if animals didn't have a sophisticated system – the optokinetic reflex – to stabilise their vision while mobile, this smeary perspective would be our permanent reality. Céline Gravot, from the Ludwig-Maximilians-Universität München, Germany, explains that most animals swivel their eyes in their heads to counteract the effects of motion: ‘You can experience this reflex when looking out of the window of a moving train; your eyes will move to follow the movement of the scene outside’, she says. But Gravot and her colleagues Alexander Knorr, Stefan Glasauer and Hans Straka were curious to find out how the swivelling motion of the eye responds to specific features of a scene. For example, would the reflex movement respond in the same way to dark objects – such as a flock of birds or squadron of planes – silhouetted against bright daylight as it does to brilliant stars against a dark sky?As the vision of most mammals is too sophisticated to begin unravelling the answer to this question, the team focused on a simpler animal, the African clawed frog (Xenopus laevis) tadpole, which swivels its eyes to counteract the effect of its swimming motion. Designing a series of simple scenes – ranging from white dots on a black background, through various shades of grey, to black dots on a white background, in addition to crescent-shaped dots and small blocks – Gravot and Knorr swivelled the scenes before the eyes of stationary tadpoles while they filmed the animals’ eye motions and recorded the electrical signals produced by the tadpoles’ retinas.However, the team was surprised when they realised that the tadpoles responded more strongly to the bright dots on the dark background than they did to the dark dots on a white background, swivelling their eyes through a wider angle in response to the white dots than the dark dots. In addition, the strength of the electrical signals generated by the retina also declined as the background became lighter and the dots darker. ‘We did not expect that’, says Gravot, as the only difference between the two scenes was the intensity of the spots relative to the background. The retina responded differently to both scenarios, ultimately altering the strength of the tadpoles swivelling reflex response.‘This confirms again that the eye is a pretty smart sensory organ on its own’, says Gravot, who suggests that the eye may be making a rudimentary interpretation of the motion of a scene to produce the reflex. And she also suspects that the tadpoles responded less strongly to the motion of dark objects because they could be interpreted as food particles, where as bright objects moving against a dark background could be the result of visual motion generated by the tadpoles’ own movements, triggering a strong visual reflex in the animals. Gravot and her colleagues are now keen to discover how the tadpoles’ drastic lifestyle change, when they metamorphose from aquatic larvae to amphibious hunters, affects their visual processing.

Developmental changes in spinal neuronal properties, motor network configuration, and neuromodulation at free-swimming stages of Xenopus tadpoles.

We describe a novel preparation of the isolated brain stem and spinal cord from prometamorphic tadpole stages of the South African clawed frog ( Xenopus laevis) that permits whole cell patch-clamp recordings from neurons in the ventral spinal cord. Previous research on earlier stages of the same species has provided one of the most detailed understandings of the design and operation of a central pattern generator circuit. Here we have addressed how development sculpts complexity from this more basic circuit. The preparation generates bouts of fictive swimming activity either spontaneously or in response to electrical stimulation of the optic tectum, allowing an investigation into how the neuronal properties, activity patterns, and neuromodulation of locomotor rhythm generation change during development. We describe an increased repertoire of cellular responses compared with younger larval stages and investigate the cellular-level effects of nitrergic neuromodulation as well as the development of a sodium pump-mediated ultraslow afterhyperpolarization (usAHP) in these free-swimming larval animals. NEW & NOTEWORTHY A novel in vitro brain stem-spinal cord preparation is described that enables whole cell patch-clamp recordings from spinal neurons in prometamorphic Xenopus tadpoles. Compared with the well-characterized earlier stages of development, spinal neurons display a wider range of firing properties during swimming and have developed novel cellular properties. This preparation now makes it feasible to investigate in detail spinal central pattern generator maturation during the dramatic switch between undulatory and limb-based locomotion strategies during amphibian metamorphosis.

Overland movement in African clawed frogs (Xenopus laevis): empirical dispersal data from within their native range.

Dispersal forms are an important component of the ecology of many animals, and reach particular importance for predicting ranges of invasive species. African clawed frogs (Xenopus laevis) move overland between water bodies, but all empirical studies are from invasive populations with none from their native southern Africa. Here we report on incidents of overland movement found through a capture-recapture study carried out over a three year period in Overstrand, South Africa. The maximum distance moved was 2.4 km with most of the 91 animals, representing 5% of the population, moving ∼150 m. We found no differences in distances moved by males and females, despite the former being smaller. Fewer males moved overland, but this was no different from the sex bias found in the population. In laboratory performance trials, we found that males outperformed females, in both distance moved and time to exhaustion, when corrected for size. Overland movement occurred throughout the year, but reached peaks in spring and early summer when temporary water bodies were drying. Despite permanent impoundments being located within the study area, we found no evidence for migrations of animals between temporary and permanent water bodies. Our study provides the first dispersal kernel for X. laevis and suggests that it is similar to many non-pipid anurans with respect to dispersal.

Does restricted access limit management of invasive urban frogs?

Management recommendations that target urban invaders should consider environmental and socio-economic aspects peculiar to the urban landscape. Urbanization often leads to the fragmentation of the invaded landscape into subunits inaccessible to managers (restricted access) or for which detailed information is lacking. Using models to explore impact of these limitations on management success provides a useful approach to propose effective countermeasures. Here we deploy a spatially explicit age-structured model applied to a pond network to investigate how restricted access and lack of detailed information may affect management of three invasive anuran species across a peri-urban landscape. The target species, the guttural toad Sclerophrys gutturalis, the African clawed frog Xenopus laevis and the painted reed frog Hyperolius marmoratus, belong to different ecotypes (terrestrial, aquatic and arboreal, respectively) and have different life history traits. We show that restricted property access significantly constrains management success in two of the three species (the guttural toad and the painted reed frog), while lack of detailed information around the invaded landscape impedes successful management in only one species (the guttural toad). The species-dependent response we detected is due to contrasting demographic and spatial invasion dynamics linked to the different anuran ecotypes. Our work highlights the necessity to adopt a context-dependent approach when proposing management recommendations in urban environment.

Dioxin Exposure Alters Molecular and Morphological Responses to Thyroid Hormone in Xenopus laevis Cultured Cells and Prometamorphic Tadpoles.

Amphibian metamorphosis is driven by thyroid hormone (TH). We used prometamorphic tadpoles and a cell line of the African clawed frog (Xenopus laevis) to examine immediate effects of dioxin exposure on TH. Gene expression patterns suggest cross-talk between the thyroid hormone receptor (TR) and aryl hydrocarbon receptor (AHR) signaling pathways. In XLK-WG cells, expression of Cytochrome P450 1A6 (cyp1A6), an AHR target, was induced 1000-fold by 100 nM TCDD (2, 3, 7, 8 tetrachlorodibenzo-p-dioxin). Krüppel-Like Factor 9 (klf9), the first gene induced in a cascade of TH responses tied to metamorphosis, was upregulated over 5-fold by 50 nM triiodothyronine (T3) and 2-fold by dioxin. Co-exposure to T3 and TCDD boosted both responses, further inducing cyp1A6 by 75% and klf9 about 60%. Additional canonical targets of each receptor, including trβa and trβb (TR) and udpgt1a (AHR) responded similarly. Induction of TH targets by TCDD in XLK-WG cells predicts that exposure could speed metamorphosis. We tested this hypothesis in two remodeling events: tail resorption and hind limb growth. Resorption of ex vivo cultured tails was accelerated by 10 nM T3, while a modest increase in resorption by 100 nM TCDD lacked statistical significance. Hind limbs doubled in length over four days following 1 nM T3 treatment, but limb length was unaffected by 100 nM TCDD. TCDD co-exposure reduced the T3 effect by nearly 40%, despite TCDD induction of klf9 in whole tadpoles, alone or with T3. These results suggest that tissue-specific TCDD effects limit or reverse the increased metamorphosis rate predicted by klf9 induction.

Shared behavioral responses and predation risk of anuran larvae and adults exposed to a novel predator

Invasive species are a regional and global threat to biological diversity. In order to evaluate an invasive predator species’ potential to harm populations of native prey species, it is critical to evaluate the behavioral responses of all life stages of the native prey species to the novel predator. The invasion of the African clawed frog (Xenopus laevis) into southern California provides an opportunity to evaluate the predation risk and behavioral responses of native amphibians. We performed predation trials and explored prey behavioral responses to determine how this invasive predator may impact native amphibian populations using Pacific chorus frogs (Pseudacris regilla) as a representative native California prey species. We found that X. laevis will readily prey upon larval and adult life stages of P. regilla. Behavior trials indicated that both larval and adult P. regilla exhibit prey response behaviors and will spatially avoid the novel invasive predator. The results suggest that native anurans may have a redundant predator response in both the larval and adult life stages, which could reduce the predatory impact of X. laevis but also drive emigration of native amphibians from invaded habitat.