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the voltage sensing phosphatase (vsp) localizes to the apical membrane of kidney tubule epithelial cells.voltage-sensing phosphatases (vsps) are transmembrane proteins that couple changes in membrane potential to hydrolysis of inositol signaling lipids. vsps catalyze the dephosphorylation of phosphatidylinositol phosphates (pips) that regulate diverse aspects of cell membrane physiology including cell division, growth and migration. vsps are highly conserved among chordates, and their rna transcripts have been detected in the adult and embryonic stages of frogs, fish, chickens, mice and humans. how ...201930964862
developmental exposure to chemicals associated with unconventional oil and gas extraction alters immune homeostasis and viral immunity of the amphibian xenopus.although aquatic vertebrates and humans are increasingly exposed to water pollutants associated with unconventional oil and gas extraction (uog), the long-term effects of these pollutants on immunity remains unclear. we have established the amphibian xenopus laevis and the ranavirus frog virus 3 (fv3) as a reliable and sensitive model for evaluating the effects of waterborne pollutants. x. laevis tadpoles were exposed to a mixture of equimass amount of uog chemicals with endocrine disrupting act ...201930939317
developmental regulation of wnt signaling by nagk and the udp-glcnac salvage pathway.in a screen for human kinases that regulate xenopus laevis embryogenesis, we identified nagk and other components of the udp-glcnac glycosylation salvage pathway as regulators of anteroposterior patterning and wnt signaling. we find that the salvage pathway does not affect other major embryonic signaling pathways (fgf, tgfβ, notch, or shh), thereby demonstrating specificity for wnt signaling. we show that the role of the salvage pathway in wnt signaling is evolutionarily conserved in zebrafish a ...201930904594
stage-dependent cardiac regeneration in xenopus is regulated by thyroid hormone availability.despite therapeutic advances, heart failure is the major cause of morbidity and mortality worldwide, but why cardiac regenerative capacity is lost in adult humans remains an enigma. cardiac regenerative capacity widely varies across vertebrates. zebrafish and newt hearts regenerate throughout life. in mice, this ability is lost in the first postnatal week, a period physiologically similar to thyroid hormone (th)-regulated metamorphosis in anuran amphibians. we thus assessed heart regeneration in ...201930755533
ketone body modulation of ligand-gated ion channels.ketogenesis is a metabolic process wherein ketone bodies are produced from the breakdown of fatty acids. in humans, fatty acid catabolism results in the production of acetyl-coa which can then be used to synthesize three ketone bodies: acetoacetate, acetone, and β-hydroxybutyrate. ketogenesis occurs at a higher rate in situations of low blood glucose, such as during fasting, heavy alcohol consumption, and in situations of low insulin, as well as in individuals who follow a 'ketogenic diet' consi ...201930562540
in vitro inhibition of human nucleoside transporters and uptake of azacitidine by an isocitrate dehydrogenase-2 inhibitor enasidenib and its metabolite agi-16903.1. the present study investigated inhibitory effects of enasidenib and its metabolite agi-16903 on (a) recombinant human nucleoside transporters (hnts) in hnt-producing xenopus laevis oocytes, and (b) azacitidine uptake in a normal b-lymphoblast peripheral blood cell line (pbc) and acute myeloid leukemia (aml) cell lines. 2. enasidenib inhibited hent1, hent2, hent3, and hent4 in oocytes with ic50 values of 7, 63, 27, and 76 μm, respectively, but exhibited little inhibition of hcnt1-3. agi-16903 ...201930394160
paracetamol-induced liver injury modelled in xenopus laevis embryos.failure to predict drug-induced liver injury (dili) remains a major contributing factor to lead compound drop-out during drug development. xenopus embryos are amenable for early stage medium throughput small molecule screens and so have the potential to be used in pre-clinical screens. to begin to assess the usefulness and limitations of xenopus embryos for safety assessment in the early phases of drug development, paracetamol was used as a model hepatotoxin. paracetamol overdose is associated w ...201930282005
conservation and divergence of protein pathways in the vertebrate heart.heart disease is the leading cause of death in the western world. attaining a mechanistic understanding of human heart development and homeostasis and the molecular basis of associated disease states relies on the use of animal models. here, we present the cardiac proteomes of 4 model vertebrates with dual circulatory systems: the pig (sus scrofa), the mouse (mus musculus), and 2 frogs (xenopus laevis and xenopus tropicalis). determination of which proteins and protein pathways are conserved and ...201931490923
discovery of the arp2 protein as a determining molecule in tumor cell death.cancer is a multifactorial disease that constitutes a serious public health problem worldwide. prostate cancer advanced stages are associated with the development of androgen-independent tumors and an apoptosis-resistant phenotype that progresses to metastasis. by studying androgen-independent lymphoid nodule carcinoma of the prostate (lncap) cells induced to apoptosis by serum elimination, we identified the activation of a non-selective cationic channel of 23ps conductance that promotes incomin ...201932091029
descubrimiento de la proteína reguladora de apoptosis 2 como determinante en la muerte de células tumorales.cancer is a multifactorial disease that constitutes a serious public health problem worldwide. prostate cancer advanced stages are associated with the development of androgen-independent tumors and an apoptosis-resistant phenotype that progresses to metastasis. by studying androgen-independent lymphoid nodule carcinoma of the prostate (lncap) cells induced to apoptosis by serum elimination, we identified the activation of a non-selective cationic channel of 23ps conductance that promotes incomin ...201931695224
a flipr assay for discovery of gabaa receptor modulators of natural origin.a fluorometric imaging plate reader (flipr) assay utilizing chinese hamster ovary (cho) cells stably transfected with gabaa receptors of α1β2γ2 subunit composition was evaluated and validated for rapid screening of plant extract libraries and efficient localization of active compounds in extracts. validation was performed with pure compounds and extracts known to contain allosteric gabaa receptor modulators. plants extracts that had been previously reported as active in an assay using xenopus la ...201931127604
loc496300 is expressed in the endoderm of developing xenopus laevis embryos. 201932550462
using hdr and a template to introduce an in-frame ha tag on the 3' end of the xenopus laevis gata2.l open reading frame. 201932550411
heterologous expression of aedes aegypti cation chloride cotransporter 2 (aeccc2) in xenopus laevis oocytes induces an enigmatic na⁺/li⁺ conductance.the yellow fever mosquito aedes aegypti possesses three genes encoding putative na⁺-coupled cation chloride cotransporters (cccs): aenkcc1, aeccc2, and aeccc3. to date, none of the aecccs have been functionally characterized. here we expressed aeccc2 heterologously in xenopus oocytes and measured the uptake of li⁺ (a tracer for na⁺) and rb⁺ (a tracer for k⁺). compared to control (h₂o-injected) oocytes, the aeccc2-expressing oocytes exhibited significantly greater uptake of li⁺, but not rb⁺. howe ...201930875796
the effects of insecticides on two splice variants of the glutamate-gated chloride channel receptor of the major malaria vector, anopheles gambiae.between half to 1 million people die annually from malaria. anopheles gambiae mosquitoes are major malaria vectors. unfortunately, resistance has emerged to the agents currently used to control a. gambiae, creating a demand for novel control measures. the pentameric glutamate-gated chloride channel (glucl) expressed in the muscle and nerve cells of these organisms are a potentially important biological target for malaria control. the pharmacological properties of anophiline glucl receptors are, ...201931479507
the role of ambient temperature and body mass on body temperature, standard metabolic rate and evaporative water loss in southern african anurans of different habitat specialisation.temperature and water availability are two of the most important variables affecting all aspects of an anuran's key physiological processes such as body temperature (tb), evaporative water loss (ewl) and standard metabolic rate (smr). since anurans display pronounced sexual dimorphism, evidence suggests that these processes are further influenced by other factors such as vapour pressure deficit (vpd), sex and body mass (mb). however, a limited number of studies have tested the generality of thes ...201931660269
environmental oxygen exposure allows for the evolution of interdigital cell death in limb patterning.amphibians form fingers without webbing by differential growth between digital and interdigital regions. amniotes, however, employ interdigital cell death (icd), an additional mechanism that contributes to a greater variation of limb shapes. here, we investigate the role of environmental oxygen in the evolution of icd in tetrapods. while cell death is restricted to the limb margin in amphibians with aquatic tadpoles, eleutherodactylus coqui, a frog with terrestrial-direct-developing eggs, has ce ...201931204171
transcriptome profiling reveals male- and female-specific gene expression pattern and novel gene candidates for the control of sex determination and gonad development in xenopus laevis.xenopus laevis is an amphibian (frog) species widely used in developmental biology and genetics. to unravel the molecular machinery regulating sex differentiation of xenopus gonads, we analyzed for the first time the transcriptome of developing amphibian gonads covering sex determination period. we applied microarray at four developmental stages: (i) nf50 (undifferentiated gonad during sex determination), (ii) nf53 (the onset of sexual differentiation of the gonads), (iii) nf56 (sexual different ...201930972573
expression of tcr genes in adult and larval xenopus laevis.in order to better understand the development and function of γδ t cells in xenopus frogs, it is necessary to determine where and when γδ t cells are found in xenopus tissues. this study examined the expression of tcr genes, focused primarily on tcr γ, in tissues of adult and larval xenopus laevis and provide new data about the expression pattern of these different tcr genes in this anuran amphibian. tcr gene expression was detected by rt-pcr in adult frog tissues including the thymus, spleen, s ...201930738793
preventing ethanol-induced brain and eye morphology defects using optogenetics.background: embryonic exposure to the teratogen ethanol leads to dysmorphias, including eye and brain morphology defects associated with fetal alcohol spectrum disorder (fasd). exposure of xenopus laevis embryos to ethanol leads to similar developmental defects, including brain and eye dysmorphism, confirming our work and the work of others showing xenopus as a useful system for studies of the brain and eye birth defects associated with fasd. several targets of ethanol action have been hypothesi ...201932685918
microsampling capillary electrophoresis mass spectrometry enables single-cell proteomics in complex tissues: developing cell clones in live xenopus laevis and zebrafish embryos.label-free single-cell proteomics by mass spectrometry (ms) is currently incompatible with complex tissues without requiring cell culturing, single-cell dissection, or tissue dissociation. we here report the first example of label-free single-cell ms-based proteomics directly in single cells in live vertebrate embryos. our approach integrates optically guided in situ subcellular capillary microsampling, one-pot extraction-digestion of the collected proteins, peptide separation by capillary elect ...201930827088
conditional demyelination and remyelination in a transgenic xenopus laevis.multiple sclerosis (ms) is the first cause of acquired disability progression in the young adult. pathology of ms associates inflammation, demyelination, and neurodegeneration. the development of immunotherapies, by reducing the relapse rate, has profoundly impacted short-term prognosis and patients' quality of life. these anti-inflammatory medications, however, have not proven to be sufficient to prevent long-term disability progression, resulting from axonal transection and neuronal damage, co ...201930820903
the c-terminus of the retinal homeobox (rax) gene product modulates transcription in a context-dependent manner.the evolutionarily conserved retinal homeobox (rax) transcription factor is essential for normal eye development in all vertebrates. despite rax's biologic significance, the molecular mechanisms underlying rax molecular function as a transcriptional regulator are poorly defined. the rax gene encodes a conserved octapeptide motif (op) near the n-terminus and several conserved regions in the c-terminus of unknown function, including the orthopedia, aristaless, rax (oar) domain and the rx domain. t ...201930820152
more than just a bandage: closing the gap between injury and appendage regeneration.the remarkable regenerative capabilities of amphibians have captured the attention of biologists for centuries. the frogs xenopus laevis and xenopus tropicalis undergo temporally restricted regenerative healing of appendage amputations and spinal cord truncations, injuries that are both devastating and relatively common in human patients. rapidly expanding technological innovations have led to a resurgence of interest in defining the factors that enable regenerative healing, and in coupling thes ...201930800076
the wnt inhibitor dkk1 is required for maintaining the normal cardiac differentiation program in xenopus laevis.wnt proteins can activate different intracellular signaling pathways. these pathways need to be tightly regulated for proper cardiogenesis. the canonical wnt/β-catenin inhibitor dkk1 has been shown to be sufficient to trigger cardiogenesis in gain-of-function experiments performed in multiple model systems. loss-of-function studies however did not reveal any fundamental function for dkk1 during cardiogenesis. using xenopus laevis as a model we here show for the first time that dkk1 is required f ...201930797757
fluorescent analogues of bekm-1 with high and specific activity against the herg channel.peptidic toxins that target specifically mammalian channels and receptors can be found in the venom of animals. these toxins are rarely used directly as tools for biochemical experiments, and need to be modified via the attachment of chemical groups (e.g., radioactive or fluorescent moieties). ideally, such modifications should maintain the toxin specificity and affinity for its target. with the goal of obtaining fluorescent derivatives of bekm-1, a toxin from the scorpion species buthus eupeus ...201932550567
rho flares repair local tight junction leaks.tight junctions contribute to epithelial barrier function by selectively regulating the quantity and type of molecules that cross the paracellular barrier. experimental approaches to evaluate the effectiveness of tight junctions are typically global, tissue-scale measures. here, we introduce zinc-based ultrasensitive microscopic barrier assay (znumba), which we used in xenopus laevis embryos to visualize short-lived, local breaches in epithelial barrier function. these breaches, or leaks, occur ...201930773490
activity and intracellular localization of senescence-associated β-galactosidase in aging xenopus oocytes and eggs.senescence-associated β-galactosidase (sa-β-gal) serves as a marker of senescence in aging somatic cells. however, little is known about sa-β-gal dynamics in aging gamete cells. to address this issue, here we investigated activity and intracellular localization of sa-β-gal in freshly obtained and aging oocytes and eggs of the african clawed frog xenopus laevis. data base mining revealed the presence of several homologous β-galactosidase sequences in the annotated xenopus genome. some of them wer ...201930769028
on the mechanism of the electrophysiological changes and membrane lesions induced by asbestos fiber exposure in xenopus laevis oocytes.the so-called amphibole asbestos fibers are enriched with mineral iron ions, able to stimulate ros production. we recently reported that crocidolite asbestos was able to interact with the cell membranes of xenopus laevis oocytes, to alter their electrical membrane properties. here, we found that applied iron ions (fe3+) or h2o2 (for ros generation) mimicked these effects, suggesting that at least one effect of iron-containing asbestos fiber exposure was mediated by ros production. furthermore, c ...201930765791
wgddetector: a pipeline for detecting whole genome duplication events using the genome or transcriptome annotations.with the availability of well-assembled genomes of a growing number of organisms, identifying the bioinformatic basis of whole genome duplication (wgd) is a growing field of genomics. the most extant software for detecting footprints of wgds has been restricted to a well-assembled genome. however, the massive poor quality genomes and the more accessible transcriptomes have been largely ignored, and in theoretically they are also likely to contribute to detect wgd using ds based method. here, to ...201930760221
various nuclear reprogramming systems using egg and oocyte materials.maternal factors stored in eggs and oocytes are necessary for reprogramming sperm for embryonic development. this reprogramming activity of maternal factors also works towards somatic cells, including terminally differentiated cells. several different experimental systems utilizing egg and oocyte materials have been applied to study nuclear reprogramming by maternal factors. among these systems, the most widely used is the transfer of a somatic cell nucleus to an oocyte arrested at the metaphase ...201930745498
pymetrozine activates trpv channels of brown planthopper nilaparvata lugens.the commercial insecticide pymetrozine has been extensively used for brown planthopper control in east asia. the transient receptor potential vanilloid (trpv) channel, which consists of two proteins, nanchung (nan) and inactive (iav), has recently been shown to be the molecular target of pymetrozine in the fruit fly (drosophila melanogaster) and pea aphid (acyrthosiphon pisum). in this study, we characterized the nan and iav trpv channel subunits of n. lugens and measured the action of pymetrozi ...201930744899
jaburetox, a natural insecticide derived from jack bean urease, activates voltage-gated sodium channels to modulate insect behavior.jaburetox (jbtx) is an insecticidal peptide derived from canavalia ensiformis urease, whose mechanism of action is not completely elucidated. we employed behavioral, electromyographical and electrophysiological protocols to identify the cellular and molecular targets involved in the jbtx entomotoxicity in cockroaches and locusts. in nauphoeta cinerea, jbtx (32 μg/g) altered the locomotory behaviour inducing a significative decrease in the distance travelled followed by a significant increase in ...201930744898
gene expression program underlying tail resorption during thyroid hormone-dependent metamorphosis of the ornamented pygmy frog microhyla fissipes.thyroid hormone (t3) is essential for vertebrate development, especially during the so-called postembryonic development, a period around birth in mammals when plasma t3 level peaks and many organs mature into their adult form. compared to embryogenesis, postembryonic development is poorly studied in mammals largely because of the difficulty to manipulate the uterus-enclosed embryos and neonates. amphibian metamorphosis is independent of maternal influence and can be easily manipulated for molecu ...201930740088
generation of xenopus haploid, triploid, and hybrid embryos.frog species of the genus xenopus are widely used for studies of cell and developmental biology, and recent genome sequencing has revealed interesting phylogenetic relationships. here we describe methods to generate haploid, triploid, and hybrid species starting from eggs and sperm of xenopus laevis and xenopus tropicalis that enable investigation of how genome size and content affect physiology at the organismal, cellular, and subcellular levels.201930737699
methods for isolating the balbiani body/germplasm from xenopus laevis oocytes.the balbiani body (bb) is a large membrane-less organelle, densely packed with mitochondria, endoplasmic reticulum, proteins, and rna. the bb is present in many vertebrate female gametes. in frogs, the bb is established early during oogenesis and operates as a maternal inherited embryonic determinant that specifies germline identity through the formation of germplasm. we describe here two techniques to isolate the bb/germplasm from xenopus laevis primary oocytes.201930737696
membrane microdomains as platform to study membrane-associated events during oogenesis, meiotic maturation, and fertilization in xenopus laevis.studies on the egg plasma membrane-associated tyrosine kinase src have shed light on the identity of the molecular machinery that is responsible for gamete interaction and possibly fusion in african clawed frog xenopus laevis. here we describe our protocol for identifying and analyzing molecular and cellular machinery that contributes to a variety of biological processes in the course of oogenesis, oocyte maturation, egg fertilization, and early embryogenesis in xenopus. our current special inte ...201930737686
reconstitution of intracellular calcium signaling in xenopus egg extracts.generation of calcium signal in the cytoplasm of fertilized or parthenogenetically activated eggs has been extensively studied in the intact eggs of several biological species. calcium transient was found to elicit a plethora of biochemical and cellular events in these cells. remarkably, intracellular calcium signaling can also be reconstituted in cell-free environment. in this chapter, we describe the methods that allow reconstitution, detection, and quantification of the calcium signal in cell ...201930737685
imatinib uptake into cells is not mediated by organic cation transporters oct1, oct2, or oct3, but is influenced by extracellular ph.cancer cells undergo genetic and environmental changes that can alter cellular disposition of drugs, notably by alterations of transmembrane drug transporters expression. whether the influx organic cation transporter 1 (oct1) encoded by the gene slc221a1 is implicated in the cellular uptake of imatinib is still controversial. besides, imatinib ionization state may be modulated by the hypoxic acidic surrounding extracellular microenvironment.201930734690
comparison the sensitivity of amphibian metamorphosis assays with nf 48 stage and nf 51 stage xenopus laevis tadpoles.the amphibian metamorphosis assay (ama) was proposed by the organization for economic cooperation and development (oecd) to screen thyroid disruptors of vertebrate species. the general experimental design of the ama exposes nieuwkoop and faber (nf) stage 51 xenopus laevis tadpoles to test chemical concentrations for 21 d. however, recent studies demonstrated that thyroid gland began to function after nf stage 45 in x. laevis. thus, in this study, we initiated exposure with nf stage 48 tadpoles w ...201930732517
using xenopus laevis oocytes to functionally characterize plant transporters.functionally characterizing plant membrane transport proteins is challenging. typically, heterologous systems are used to study them. immature eggs (oocytes) of the south african clawed frog xenopus laevis are considered an ideal expression system for such studies. these large oocytes have a low number of endogenous transport systems in their plasma membranes and highly express foreign mrna; the oocyte plasma membrane is the default destination of integral membrane proteins that lack recognized ...201930707001
anillin regulates epithelial cell mechanics by structuring the medial-apical actomyosin network.cellular forces sculpt organisms during development, while misregulation of cellular mechanics can promote disease. here, we investigate how the actomyosin scaffold protein anillin contributes to epithelial mechanics in xenopus laevis embryos. increased mechanosensitive recruitment of vinculin to cell-cell junctions when anillin is overexpressed suggested that anillin promotes junctional tension. however, junctional laser ablation unexpectedly showed that junctions recoil faster when anillin is ...201930702429
menthol acts as a positive allosteric modulator on nematode levamisole sensitive nicotinic acetylcholine receptors.the ongoing and widespread emergence of resistance to the existing anti-nematodal pharmacopeia has made it imperative to develop new anthelminthic agents. historically, plants have been important sources of therapeutic compounds and offer an alternative to synthetic drugs. monoterpenoids are phytochemicals that have been shown to produce acute toxic effects in insects and nematodes. previous studies have shown nicotinic acetylcholine receptors (nachrs) to be possible targets for naturally occurr ...201930682641
characterization of xenopus laevis guanine deaminase reveals new insights for its expression and function in the embryonic kidney.the mammalian guanine deaminase (gda), called cypin, is important for proper neural development, by regulating dendritic arborization through modulation of microtubule (mt) dynamics. additionally, cypin can promote mt assembly in vitro. however, it has never been tested whether cypin (or other gda orthologs) binds to mts or modulates mt dynamics. here, we address these questions and characterize xenopus laevis gda (gda) for the first time during embryonic development.201930682232
analysis of long non-coding rna involved in atrazine-induced testicular degeneration of xenopus laevis.long non-coding rna (lncrna) plays a critical role in male germline development. atrazine (az) as an environmental endocrine disrupting chemical (edcs) can induce male reproductive toxicity in amphibians. our previous studies demonstrated that az can alter gene and circular rna (circrna) expression of damaged testes in xenopus laevis (x. laevis). we furthered to investigate the lncrna expression profiling in the testis of x. laevis. over 3559 lncrnas were detected by lncrna sequencing. az induce ...201930675760
oscillating aquaporin phosphorylation and 14-3-3 proteins mediate the circadian regulation of leaf hydraulics.the circadian clock regulates plant tissue hydraulics to synchronize water supply with environmental cycles and thereby optimize growth. the circadian fluctuations in aquaporin transcript abundance suggest that aquaporin water channels play a role in these processes. here, we show that hydraulic conductivity (kros) of arabidopsis (arabidopsis thaliana) rosettes displays a genuine circadian rhythmicity with a peak around midday. combined immunological and proteomic approaches revealed that phosph ...201930674691
non-acylated wnts can promote signaling.wnts are a family of 19 extracellular ligands that regulate cell fate, proliferation, and migration during metazoan embryogenesis and throughout adulthood. wnts are acylated post-translationally at a conserved serine and bind the extracellular cysteine-rich domain (crd) of frizzled (fzd) seven-pass transmembrane receptors. although crystal structures suggest that acylation is essential for wnt binding to fzds, we show here that several wnts can promote signaling in xenopus laevis and danio rerio ...201930673610
effect of the flavonoid hesperidin on glucose and fructose transport, sucrase activity and glycaemic response to orange juice in a crossover trial on healthy volunteers.although polyphenols inhibit glucose absorption and transport in vitro, it is uncertain whether this activity is sufficient to attenuate glycaemic response in vivo. we examined this using orange juice, which contains high levels of hesperidin. we first used a combination of in vitro assays to evaluate the potential effect of hesperidin and other orange juice components on intestinal sugar absorption and then tested whether this translated to an effect in healthy volunteers. hesperidin attenuated ...201930670104
whole-cell photoacoustic sensor based on pigment relocalization.photoacoustic (optoacoustic) imaging can extract molecular information with deeper tissue penetration than possible by fluorescence microscopy techniques. however, there is currently still a lack of robust genetically controlled contrast agents and molecular sensors that can dynamically detect biological analytes of interest with photoacoustics. in a biomimetic approach, we took inspiration from cuttlefish who can change their color by relocalizing pigment-filled organelles in so-called chromato ...201930663315
x-linked α-thalassemia with mental retardation is downstream of protein kinase a in the meiotic cell cycle signaling cascade in xenopus oocytes and is dynamically regulated in response to dna damage†.x-linked α-thalassemia with mental retardation (atrx) is a chromatin remodeling protein that belongs to the switch/sucrose non-fermentable (swi2/snf2) family of helicase/atpases. during meiosis, atrx is necessary for heterochromatin formation and maintenance of chromosome stability in order to ensure proper assembly of the metaphase ii spindle. previously, we established atrx as a novel progesterone regulated protein during bovine meiotic maturation, in addition to being dynamically regulated in ...201930649195
the electrical block to polyspermy induced by an intracellular ca2+ increase at fertilization of the clawed frogs, xenopus laevis and xenopus tropicalis.polyspermy blocking, to ensure monospermic fertilization, is necessary for normal diploid development in most animals. we have demonstrated here that monospermy in the clawed frog, xenopus tropicalis, as well as in x. laevis, is ensured by a fast, electrical block to polyspermy on the egg plasma membrane after the entry of the first sperm, which is mediated by the positive-going fertilization potential. an intracellular ca2+ concentration ([ca2+ ]i ) at the sperm entry site was propagated as a c ...201930648313
xenopus oocytes as a heterologous expression system for analysis of tight junction proteins.claudins (cldns) represent the largest family of transmembrane tight junction (tj) proteins, determining organ-specific epithelial barrier properties. because methods for the analysis of multiple cldn interaction are limited, we have established the heterologous xenopus laevis oocyte expression system for tj protein assembly and interaction analysis. oocytes were injected with crna encoding human cldn-1, -2, or -3 or with a combination of these and were incubated in pairs for interaction analysi ...201930645152
insemination or phosphatidic acid induces an outwardly spiraling disk of elevated ca2+ to produce the ca2+ wave during xenopus laevis fertilization.during xenopus fertilization, the initial intracellular calcium ((ca2+)i) release at the sperm-egg binding site (hot spot) has not been described without the use of inhibitors, nor related to underlying er structure. without inhibitors, we now report that sperm induce an initial hot spot after sperm addition to xenopus eggs that was ~25 µm. this area is consistent with the size of er patches and clusters of ip3 receptors that have enhanced activity. furthermore, we find a new mechanism for the f ...201930641042
deuterium-free, three-plexed peptide diethylation for highly accurate quantitative proteomics.the deuterium, a frequently used stable isotope in isotopic labeling for quantitative proteomics, could deteriorate the accuracy and precision of proteome quantification owing to the retention time shift of deuterated peptides from the hydrogenated counterpart. we introduce a novel three-plexed peptide "diethylation" using only 13c isotopologues of acetaldehyde and demonstrate that the accuracy and precision of our method in proteome quantification are significantly superior to the conventional ...201930638020
purification and characterization of a urea sensitive lactate dehydrogenase from skeletal muscle of the african clawed frog, xenopus laevis.the african clawed frog, xenopus laevis endures whole body dehydration which can increase its reliance on anaerobic glycolysis for energy production. this makes the regulation of the terminal enzyme of glycolysis, lactate dehydrogenase (ldh), crucial to stress survival. we investigated the enzymatic properties and posttranslational modification state of purified ldh from the skeletal muscle of control and dehydrated (30% total body water loss) x. laevis. ldh from the muscle of dehydrated frogs s ...201930631901
rhinella marina oocytes: a suitable alternative expression system for functional characterization of aquaglyceroporins.amphibian oocytes have been extensively used for heterologous expression of membrane proteins for studying their biochemical and biophysical properties. so far, xenopus laevis is the main amphibian used as oocytes source to express aquaglyceroporins in order to assess water and solutes permeability. however, this well-established amphibian model represents a threat to the biodiversity in many countries, especially in those from tropical regions. for that reason, the import of xenopus laevis is s ...201930631140
identification of vacuolar phosphate efflux transporters in land plants.inorganic phosphate (pi) is an essential component of all life forms. land plants acquire pi from the soil through roots and associated symbioses, and it is then transported throughout the plant. when sufficient, excess pi is stored in vacuoles for remobilization following pi deficiency. although pi release from the vacuoles to the cytoplasm serves as a critical mechanism for plants to adapt to low-pi stress, the transporters responsible for vacuolar pi efflux have not been identified. here, we ...201930626920
ensa and arpp19 differentially control cell cycle progression and development.greatwall (gwl) is an essential kinase that indirectly controls pp2a-b55, the phosphatase counterbalancing cyclin b/cdk1 activity during mitosis. in xenopus laevis egg extracts, gwl-mediated phosphorylation of overexpressed arpp19 and ensa turns them into potent pp2a-b55 inhibitors. it has been shown that the gwl/ensa/pp2a-b55 axis contributes to the control of dna replication, but little is known about the role of arpp19 in cell division. by using conditional knockout mouse models, we investiga ...201930626720
xenopus slc7a5 is essential for notochord function and eye development.slc7a5 (also known as lat1), largely accepted as an amino acid transporter, has been shown to play important roles in cancer and developmental processes. because knockout mice of slc7a5 are embryonically lethal due to placental defects, it is difficult to evaluate its role in early development. in this study, expression and function of slc7a5 were evaluated in xenopus laevis embryos that develop without a placenta. expression of slc7a5 was detected in the notochord and in the eye and it was not ...201930625369
reinterpretation of the substrate specificity of the voltage-sensing phosphatase during dimerization.voltage-sensing phosphatases (vsps) cleave both 3- and 5-phosphates from inositol phospholipids in response to membrane depolarization. when low concentrations of ciona intestinalis vsp are expressed in xenopus laevis oocytes, the 5-phosphatase reaction can be observed during large membrane depolarizations. when higher concentrations are expressed, the 5-phosphatase activity is observed with smaller depolarizations, and the 3-phosphatase activity is revealed with strong depolarization. here we a ...201930622132
probing the dynamics and structural topology of the reconstituted human kcnq1 voltage sensor domain (q1-vsd) in lipid bilayers using electron paramagnetic resonance spectroscopy.kcnq1 (kv7.1 or kvlqt1) is a potassium ion channel protein found in the heart, ear, and other tissues. in complex with the kcne1 accessory protein, it plays a role during the repolarization phase of the cardiac action potential. mutations in the channel have been associated with several diseases, including congenital deafness and long qt syndrome. nuclear magnetic resonance (nmr) structural studies in detergent micelles and a cryo-electron microscopy structure of kcnq1 from xenopus laevis have s ...201930620191
immunohistochemical localization of calpains in the amphibian xenopus laevis.though histochemical techniques have been used for decades, they are still very important in basic research. they make it possible to work on fixed tissues and provide a large amount of information in a relatively short time and at a low cost. here we describe methods for indirect immunohistochemistry and immunofluorescence on sections of tadpoles and tissues of adult amphibians belonging to the species xenopus laevis. the objective is to localize calpains within tissues in order to understand t ...201930617797
a stable tetramer is not the only oligomeric state that mitochondrial single-stranded dna binding proteins can adopt.mitochondrial single-stranded dna (ssdna)-binding proteins (mtssbs) are required for mitochondrial dna replication and stability and are generally assumed to form homotetramers, and this species is proposed to be the one active for ssdna binding. however, we recently reported that the mtssb from saccharomyces cerevisiae (scrim1) forms homotetramers at high protein concentrations, whereas at low protein concentrations, it dissociates into dimers that bind ssdna with high affinity. in this work, u ...201930617184
arid3a regulates nephric tubule regeneration via evolutionarily conserved regeneration signal-response enhancers.amphibians and fish have the ability to regenerate numerous tissues, whereas mammals have a limited regenerative capacity. despite numerous developmental genes becoming reactivated during regeneration, an extensive analysis is yet to be performed on whether highly regenerative animals utilize unique cis-regulatory elements for the reactivation of genes during regeneration and how such cis-regulatory elements become activated. here, we screened regeneration signal-response enhancers at the lhx1 l ...201930616715
applications of in-cell nmr in structural biology and drug discovery.in-cell nuclear magnetic resonance (nmr) is a method to provide the structural information of a target at an atomic level under physiological conditions and a full view of the conformational changes of a protein caused by ligand binding, post-translational modifications or protein⁻protein interactions in living cells. previous in-cell nmr studies have focused on proteins that were overexpressed in bacterial cells and isotopically labeled proteins injected into oocytes of xenopus laevis or delive ...201930609728
curare alkaloids from matis dart poison: comparison with d-tubocurarine in interactions with nicotinic, 5-ht3 serotonin and gabaa receptors.several novel bisbenzylisoquinoline alkaloids (bbiqas) have recently been isolated from a matis tribe arrow poison and shown by two-electrode voltage-clamp to inhibit mouse muscle nicotinic acetylcholine receptors (nachr). here, using radioligand assay with aplysia californica achbp and radioiodinated α-bungarotoxin ([125i]-αbgt), we show that bbiqa1, bbiqa2, and d-tubocurarine (d-tc) have similar affinities to nachr orthosteric site. however, a competition with [125i]-αbgt for binding to the to ...201930608952
skin grafting in xenopus laevis: a technique for assessing development and immunological disparity.skin grafting in the amphibian xenopus laevis has been used to detect not only allogeneic antigens that differ by minor h antigens or by one mhc haplotype, but also to detect ontogeny-specific antigens (including both emerging adult- and disappearing larval-specific) during metamorphosis. to understand the mechanisms underlying allogeneic tolerance or immune responses against larval- and/or adult-specific antigens, a complete mhc homozygous, inbred strain is the most appropriate experimental mod ...201930606753
methodological improvements for fluorescence recordings in xenopus laevis oocytes.xenopus laevis oocytes are a widely used model system because of their capacity to translate exogenous mrna, but their high intrinsic background fluorescence is a disadvantage for fluorescence recordings. here, we developed two distinct methods for improving fluorescence recordings from oocytes. one was a pharmacological method in which a small-molecule salt-inducible kinase inhibitor was co-injected with the mrna of interest to stimulate melanin production. we interrogated the oocytes using cut ...201930606741
cdk phosphorylation of xenopus laevis m18bp1 promotes its metaphase centromere localization.chromosome segregation requires the centromere, the site on chromosomes where kinetochores assemble in mitosis to attach chromosomes to the mitotic spindle. centromere identity is defined epigenetically by the presence of nucleosomes containing the histone h3 variant cenp-a. new cenp-a nucleosome assembly occurs at the centromere every cell cycle during g1, but how cenp-a nucleosome assembly is spatially and temporally restricted remains poorly understood. centromere recruitment of factors requi ...201930606714
the cmg helicase bypasses dna-protein cross-links to facilitate their repair.covalent dna-protein cross-links (dpcs) impede replication fork progression and threaten genome integrity. using xenopus egg extracts, we previously showed that replication fork collision with dpcs causes their proteolysis, followed by translesion dna synthesis. we show here that when dpc proteolysis is blocked, the replicative dna helicase cmg (cdc45, mcm2-7, gins), which travels on the leading strand template, bypasses an intact leading strand dpc. single-molecule imaging reveals that gins doe ...201930595447
xenopus laevis fgf16 activates the expression of genes coding for the transcription factors sp5 and sp5l.fibroblast growth factors (fgfs) comprise a family of signalling molecules with essential roles in early embryonic development across animal species. the role of fgfs in mesoderm formation and patterning in xenopus has been particularly well studied. however, little is known about fgf16 in xenopus. using in situ hybridisation, we uncover the expression pattern of fgf16 during early xenopus laevis development, which has not been previously described. we show that the zygotic expression of fgf16 i ...201932149373
gd(iii)-gd(iii) relaxation-induced dipolar modulation enhancement for in-cell electron paramagnetic resonance distance determination.in-cell distance determination by electron paramagnetic resonance (epr) spectroscopy reveals essential structural information about biomacromolecules under native conditions. we demonstrate that the pulsed epr technique ridme (relaxation induced dipolar modulation enhancement) can be utilized for such distance determination. the performance of in-cell ridme has been assessed at q-band using stiff molecular rulers labeled with gd(iii)-pymta and microinjected into xenopus laevis oocytes. the overt ...201930864799
dynamin binding protein is required for xenopus laevis kidney development.the adult human kidney contains over one million nephrons, with each nephron consisting of a tube containing segments that have specialized functions in nutrient and water absorption and waste excretion. the embryonic kidney of xenopus laevis consists of a single functional nephron composed of regions that are analogous to those found in the human nephron, making it a simple model for the study of nephrogenesis. the exocyst complex, which traffics proteins to the cell membrane in vesicles via cd ...201930863317
foxn3 is necessary for the development of the interatrial septum, the ventricular trabeculae and the muscles at the head/trunk interface in the african clawed frog, xenopus laevis (lissamphibia: anura: pipidae).fox genes are a large family of transcription factors that play diverse roles in the immune system, metabolism, cancer, cell cycle, and animal development. it has been shown that foxn3 is indispensable for normal craniofacial development in the mouse and the african clawed frog, xenopus laevis. morpholino-mediated knockdown of foxn3 in x. laevis delays overall development of early tadpole stages and causes eye defects, the absence of some cranial nerve branches, and malformations of the cranial ...201930859697
mechanical force induces phosphorylation-mediated signaling that underlies tissue response and robustness in xenopus embryos.mechanical forces are essential drivers of numerous biological processes, notably during development. although it is well recognized that cells sense and adapt to mechanical forces, the signal transduction pathways that underlie mechanosensing have remained elusive. here, we investigate the impact of mechanical centrifugation force on phosphorylation-mediated signaling in xenopus embryos. by monitoring temporal phosphoproteome and proteome alterations in response to force, we discover and valida ...201930852251
a unique role of thyroid hormone receptor β in regulating notochord resorption during xenopus metamorphosis.tail resorption during anuran metamorphosis is perhaps the most dramatic tissue transformation that occurs during vertebrate development. earlier studies in highly related anuran species xenopus laevis and xenopus tropicalis have shown that thyroid hormone (t3) receptor (tr) plays a necessary and sufficient role to mediate the causative effect of t3 on metamorphosis. of the two known tr genes in vertebrates, trα is highly expressed during both premetamorphosis and metamorphosis while trβ express ...201930851299
distinctive single-channel properties of α4β2-nicotinic acetylcholine receptor isoforms.central nervous system nicotinic acetylcholine receptors (nachr) are predominantly of the α4β2 subtype. two isoforms exist, with high or low agonist sensitivity (hs-(α4β2)2β2- and ls-(α4β2)2α4-nachr). both isoforms exhibit similar macroscopic potency and efficacy values at low acetylcholine (ach) concentrations, mediated by a common pair of high-affinity α4(+)/(-)β2 subunit binding interfaces. however ls-(α4β2)2α4-nachr also respond to higher concentrations of ach, acting at a third α4(+)/(-)α4 ...201930845161
evolution of the rho guanine nucleotide exchange factors kalirin and trio and their gene expression in xenopus development.guanine nucleotide exchange factors (gefs) activate rho gtpases by accelerating their gdp/gtp exchange. trio and its paralog kalirin (kalrn) are unique members of the rho-gefs that harbor three catalytic domains: two functional gef domains and a serine/threonine kinase domain. the n-terminal gef domain activates rac1 and rhog gtpases, while the c-terminal gef domain acts specifically on rhoa. trio and kalrn have an evolutionary conserved function in morphogenetic processes including neuronal dev ...201930844509
electrophysiological changes during early steps of retinitis pigmentosa.the rhodopsin mutation p23h is responsible for a significant portion of autosomal-dominant retinitis pigmentosa, a disorder characterized by rod photoreceptor death. the mechanisms of toxicity remain unclear; previous studies implicate destabilization of p23h rhodopsin during light exposure, causing decreased endoplasmic reticulum (er) exit and er stress responses. here, we probed phototransduction in xenopus laevis rods expressing bovine p23h rhodopsin, in which retinal degeneration is inducibl ...201930840038
erratum: skin grafting in xenopus laevis: a technique for assessing development and immunological disparity. 201931371466
jmjd6a regulates gsk3β rna splicing in xenopus laevis eye development.it has been suggested that jmjd6 plays an important role in gene regulation through its demethylation or hydroxylation activity on histone and transcription factors. in addition, jmjd6 has been shown to regulate rna splicing by interaction with splicing factors. in this study, we demonstrated that jmjd6a is expressed in developing xenopus laevis eye during optic vesicle formation and retinal layer differentiation stages. knockdown of jmjd6a by an antisense morpholino resulted in eye malformation ...201931361752
identification of the catechin uptake transporter responsible for intestinal absorption of epigallocatechin gallate in mice.many studies have shown that epigallocatechin gallate (egcg) contribute to the health benefits of green tea, although its bioavailability is usually low. however, the mechanism underlying its intestinal absorption remains unclear. in human subjects, it has been reported that the bioavailability of egcg increases after repeated oral catechin intake. we hypothesized that a certain uptake transporter was involved in this increase, and investigated a novel egcg transporter. we first confirmed the in ...201931358798
maldi-imaging of early stage xenopus laevis embryos.xenopus laevis is an important model organism for vertebrate development. an extensive literature has developed on changes in transcript expression during development of this organism, and there is a growing literature on the corresponding protein expression changes during development. in contrast, there is very little information on changes in metabolite expression during development. we present the first maldi mass-spectrometry images of metabolites within the developing embryo. these images w ...201931357275
encephalopathies with kcnc1 variants: genotype-phenotype-functional correlations.to analyze clinical phenotypes associated with kcnc1 variants other than the progressive myoclonus epilepsy-causing variant p.arg320his, determine the electrophysiological functional impact of identified variants and explore genotype-phenotype-physiological correlations.201931353855
spatial analysis of reck, mt1-mmp, and timp-2 proteins during early xenopus laevis development.proper cell-cell and cell-ecm interactions are vital for cell migration and patterning of the vertebrate embryo. mmps and their inhibitors, reck and timps, are all differentially expressed during embryogenesis to regulate such ecm remodeling and cell interactions. mt1-mmp, reck, and timp-2 are unique amongst other ecm-regulating proteins as they act in the pericellular space. past studies have shown that reck and timp-2 interact with mt1-mmp on the cell surface, thereby influencing cell behaviou ...201931352120
embryogenesis of marsupial frogs (hemiphractidae), and the changes that accompany terrestrial development in frogs.the developmental adaptations of the marsupial frogs gastrotheca riobambae and flectonotus pygmaeus (hemiphractidae) are described and compared with frogs belonging to seven additional families. incubation of embryos by the mother in marsupial frogs is associated with changes in the anatomy and physiology of the female, modifications of oogenesis, and extraordinary changes in embryonic development. the comparison of early development reveals that gene expression is highly conserved. however, the ...201931598865
identification of crucial residues in α-conotoxin ei inhibiting muscle nicotinic acetylcholine receptor.α-conotoxins (α-ctxs) are small disulfide-rich peptides from venom of conus species that target nicotinic acetylcholine receptors (nachrs). the muscle-type nachrs have been recognized as a potential target for several diseases, such as myogenic disorders, muscle dystrophies, and myasthenia gravis. ei, an α4/7-ctx, mainly blocks α1β1δε nachrs and has an extra n-terminal extension of three amino acids. in this study, the alanine scanning (ala-scan) mutagenesis was applied in order to identify key ...201931623211
popdc3 gene variants associate with a new form of limb girdle muscular dystrophy.the popeye domain containing 3 (popdc3) gene encodes a membrane protein involved in cyclic adenosine monophosphate (camp) signaling. besides gastric cancer, no disease association has been described. we describe a new muscular dystrophy associated with this gene.201931610034
xenopus laevis as a bioindicator of endocrine disruptors in the region of central chile.one of the direct causes of biodiversity loss is environmental pollution resulting from the use of chemicals. different kinds of chemicals, such as persistent organic pollutants and some heavy metals, can be endocrine disruptors, which act at low doses over a long period of time and have a negative effect on the reproductive and thyroid system in vertebrates worldwide. research on the effects of endocrine disruptors and the use of bioindicators in neotropical ecosystems where pressure on biodive ...201931422435
changes in ca2+ removal can mask the effects of geometry during ip3r mediated ca2+ signals.calcium (ca2+) signals are ubiquitous. most intracellular ca2+ signals involve the release of ca2+ from the endoplasmic reticulum (er) through inositol 1,4,5-trisphosphate receptors (ip3rs). the non-uniform spatial organization of ip3rs and the fact that their individual openings are coupled via cytosolic ca2+ are key factors for the variety of spatio-temporal distributions of the cytosolic [ca2+] and the versatility of the signals. in this paper we combine experiments performed in untreated and ...201931417423
gaining insights in the nutritional metabolism of amphibians: analyzing body nutrient profiles of the african clawed frog, xenopus laevis.whole bodies of xenopus laevis (n = 19) were analysed for chemical composition and morphometrics. the nutrient profile (macronutrients, amino acids, fatty acids and minerals) was evaluated by sex; interactions among variables with body weights and lengths, and comparisons made with different species of marine and fresh water fish. significant differences were found in morphometric measurements, water content, several minerals and fatty acids between sexes of x. laevis. amino acid profiles differ ...201931410305
novel vectors for functional interrogation of xenopus orfeome coding sequences.the current xenopus orfeome contains ~10,250 validated, full-length cdna sequences without stop codons from xenopus laevis and ~3,970 from xenopus tropicalis cloned into gateway-compatible entry vectors. to increase the utility of the orfeome, we have constructed the gateway-compatible destination vectors pdxtp and pdxtr, which in combination can control the spatial and temporal expression of any open reading frame (orf). pdxtp receives a promoter/enhancer of interest, which controls the spatial ...201931403250
comparative embryonic spatio-temporal expression profile map of the xenopus p2x receptor family.p2x receptors are atp-gated cations channels formed by the homo or hetero-trimeric association from the seven cloned subunits (p2x1-7). p2x receptors are widely distributed in different organs and cell types throughout the body including the nervous system and are involved in a large variety of physiological but also pathological processes in adult mammals. however, their expression and function during embryogenesis remain poorly understood. here, we report the cloning and the comparative expres ...201931402854
radiographic and ultrasonographic appearance of pneumonia in a frog.a 12-year-old female african clawed frog (xenopus laevis) displayed lethargy and anorexia for 1 week. radiographs detected a moderately reduced volume of aerated lung with lobulated margins with the left lung predominantly affected. transcelomic ultrasound revealed marked consolidation of the left lung. postmortem examination confirmed suppurative and histiocytic pneumonia, worse in the left lung, caused by mycobacterium chelonae. given that amphibian pulmonary consolidation and celomic effusion ...201931392794
evolution of cis-regulatory modules for the head organizer gene goosecoid in chordates: comparisons between branchiostoma and xenopus.in cephalochordates (amphioxus), the notochord runs along the dorsal to the anterior tip of the body. in contrast, the vertebrate head is formed anterior to the notochord, as a result of head organizer formation in anterior mesoderm during early development. a key gene for the vertebrate head organizer, goosecoid (gsc), is broadly expressed in the dorsal mesoderm of amphioxus gastrula. amphioxus gsc expression subsequently becomes restricted to the posterior notochord from the early neurula. thi ...201931388442
stereoselective anti-cancer activities of ginsenoside rg3 on triple negative breast cancer cell models.ginsenoside rg3 (rg3) has two epimers, 20(s)-ginsenoside rg3 (srg3) and 20(r)-ginsenoside rg3 (rrg3), and while rg3 itself has been reported to have anti-cancer properties, few studies have been reported on the anti-cancer effects of the different epimers. the aim was to investigate the stereoselective effects of the rg3 epimers on triple negative breast cancer (tnbc) cell lines, tested using cell-based assays for proliferation, apoptosis, cell cycle arrest, migration and invasion. molecular doc ...201931374984
allocation trade-offs impact organ size and muscle architecture in an invasive population of xenopus laevis in western france.invasive species are a global scourge and often negatively impact native species. understanding the expansion and dispersal limits of these species is essential. as previous studies have demonstrated increased locomotor performance for populations at the edge of the range of expanding populations, studies of locomotion including the anatomical and physiological traits underlying dispersal capacity are of interest. we focus here on an invasive population of xenopus laevis introduced in france nea ...201931373390
a herbivore-induced plant volatile of the host plant acts as a collective foraging signal to the larvae of the meadow moth, loxostege sticticalis (lepidoptera: pyralidae).the meadow moth loxostege sticticalis is a serious agricultural pest that feeds on the leaves of many economic crops, such as sugar beet, soybean, sunflower, and potato. in addition to the rapid migration of adult moths, the collective foraging behavior of the larvae is also thought to be involved in the search for new food sources and substantially contributes to the expansion of the infested area. however, whether and how the chemical signals take part in this process remains unknown. in this ...201931499032
inferring the "primordial immune complex": origins of mhc class i and antigen receptors revealed by comparative genomics.comparative analyses suggest that the mhc was derived from a prevertebrate "primordial immune complex" (pic). pic duplicated twice in the well-studied two rounds of genome-wide duplications (2r) early in vertebrate evolution, generating four mhc paralogous regions (predominantly on human chromosomes [chr] 1, 6, 9, 19). examining chiefly the amphibian xenopus laevis, but also other vertebrates, we identified their mhc paralogues and mapped mhc class i, agr, and "framework" genes. most class i gen ...201931492741
pharmacological characterization of a homomeric nicotinic acetylcholine receptor formed by ancylostoma caninum acr-16.parasitic nematode infections are treated using anthelmintic drugs, some of which target nicotinic acetylcholine receptors (nachrs) located in different parasite tissues. the limited arsenal of anthelmintic agents and the prevalence of drug resistance imply that future defense against parasitic infections will depend on the discovery of novel targets and therapeutics. previous studies have suggested that ascaris suum acr-16 nachrs are a suitable target for the development of antinematodal drugs. ...201931486912
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