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evaluation of a commercial electro-kinetically pumped sheath-flow nanospray interface coupled to an automated capillary zone electrophoresis system.capillary zone electrophoresis-electrospray ionization-mass spectrometry (cze-esi-ms) is attracting renewed attention for proteomic and metabolomic analysis. an important reason for this interest is the maturation and commercialization of interfaces for coupling cze with esi-ms. one of these interfaces is an electro-kinetically pumped sheath flow nanospray interface developed by the dovichi group, in which a very low sheath flow is generated based on electroosmosis within a glass emitter. cmp sc ...201727981343
δβγ-enac is inhibited by cftr but stimulated by camp in xenopus laevis oocytes.the epithelial sodium channel (enac) and the cystic fibrosis transmembrane conductance regulator (cftr) chloride channel critically regulate airway surface liquid by driving fluid absorption and secretion, respectively. their functional interplay is complex and incompletely understood. enac is a heteromeric channel with three well-characterized subunits (α, β, and γ). in humans, an additional δ-enac subunit exists in lung and several other tissues, where it may replace the α-subunit to form δβγ- ...201727941075
controlling the messenger: regulated translation of maternal mrnas in xenopus laevis development.the selective translation of maternal mrnas encoding cell-fate determinants drives the earliest decisions of embryogenesis that establish the vertebrate body plan. this chapter will discuss studies in xenopus laevis that provide insights into mechanisms underlying this translational control. xenopus has been a powerful model organism for many discoveries relevant to the translational control of maternal mrnas because of the large size of its oocytes and eggs that allow for microinjection of mole ...201727975270
primary brain calcification causal pit2 transport-knockout variants can exert dominant negative effects on wild-type pit2 transport function in mammalian cells.primary brain calcification (pbc) is a neurodegenerative disorder characterized by calcium-phosphate deposits in the basal ganglia and often also other areas of the brain. the prevalent clinical manifestations are cognitive impairment, neuropsychiatric symptoms, and movement disorders. in recent years, monoallelic variants in slc20a2, which encodes the type iii sodium-dependent inorganic phosphate (pi) transporter 2 (pit2), have been linked to the familial form of pbc in 40-50% of the families r ...201727943094
xenopus laevis neuronal cell adhesion molecule (nrcam): plasticity of a cam in the developing nervous system.neuron-glial-related cell adhesion molecule (nrcam) is a neuronal cell adhesion molecule of the l1 immunoglobulin superfamily, which plays diverse roles during nervous system development including axon growth and guidance, synapse formation, and formation of the myelinated nerve. perturbations in nrcam function cause a wide variety of disorders, which can affect wiring and targeting of neurons, or cause psychiatric disorders as well as cancers through abnormal modulation of signaling events. in ...201727942869
evolution of vocal patterns: tuning hindbrain circuits during species divergence.the neural circuits underlying divergent courtship behaviors of closely related species provide a framework for insight into the evolution of motor patterns. in frogs, male advertisement calls serve as unique species identifiers and females prefer conspecific to heterospecific calls. advertisement calls of three relatively recently (∼8.5 mya) diverged species - xenopus laevis, x. petersii and x. victorianus - include rapid trains of sound pulses (fast trills). we show that while fast trills are ...201728011819
nemo-like kinase 1 (nlk1) and paraxial protocadherin (papc) cooperatively control xenopus gastrulation through regulation of wnt/planar cell polarity (pcp) signaling.the wnt/planar cell polarity (pcp) pathway directs cell migration during vertebrate gastrulation and is essential for proper embryonic development. paraxial protocadherin (papc, gene symbol pcdh8.2) is an important activator of wnt/pcp signaling during xenopus gastrulation, but how papc activity is controlled is incompletely understood. here we show that nemo-like kinase 1 (nlk1), an atypical mitogen-activated protein (map) kinase, physically associates with the c-terminus of papc. this interact ...201727875771
reversible developmental stasis in response to nutrient availability in the xenopus laevis central nervous system.many organisms confront intermittent nutrient restriction (nr), but the mechanisms to cope with nutrient fluctuations during development are not well understood. this is particularly true of the brain, the development and function of which is energy intensive. here we examine the effects of nutrient availability on visual system development in xenopus laevis tadpoles. during the first week of development, tadpoles draw nutrients from maternally provided yolk. upon yolk depletion, animals forage ...201727875263
thyroid hormone-induced activation of notch signaling is required for adult intestinal stem cell development during xenopus laevis metamorphosis.in xenopus laevis intestine during metamorphosis, the larval epithelial cells are removed by apoptosis, and the adult epithelial stem (ae) cells appear concomitantly. they proliferate and differentiate to form the adult epithelium (ep). thyroid hormone (th) is well established to trigger this remodeling by regulating the expression of various genes including notch receptor. to study the role of notch signaling, we have analyzed the expression of its components, including the ligands (dll and jag ...201727870267
use of xenopus laevis oocytes to study auxin transport.xenopus laevis oocytes are an expression system that is particularly well suited for the characterization of membrane transporters. oocytes possess only very little endogenous transport systems and therefore transporters can be studied with a high signal-to-noise ratio. this book chapter provides the basic methods to use xenopus oocytes for the characterization of transporters by radiotracer experiments. while the methods described here were established to study auxin transport they can easily b ...201727864772
cftr-β-catenin interaction regulates mouse embryonic stem cell differentiation and embryonic development.cystic fibrosis transmembrane conductance regulator (cftr) is a camp-regulated anion channel capable of conducting both cl- and hco3-, mutations of which cause cystic fibrosis (cf), a common autosomal recessive disease. although cf patients are known to have varied degree of developmental problems, the biological role of cftr in embryonic development remains elusive. here, we show that cftr is functionally expressed in mouse escs. cftr-/- mescs exhibit dramatic defect in mesendoderm differentiat ...201727834953
clustered xenopus keratin genes: a genomic, transcriptomic, and proteomic analysis.keratin genes belong to the intermediate filament superfamily and their expression is altered following morphological and physiological changes in vertebrate epithelial cells. keratin genes are divided into two groups, type i and ii, and are clustered on vertebrate genomes, including those of xenopus species. various keratin genes have been identified and characterized by their unique expression patterns throughout ontogeny in xenopus laevis; however, compilation of previously reported and newly ...201727842699
electrophysiological activity of a neurotoxic fraction from the venom of box jellyfish carybdea marsupialis.carybdea marsupialis is a widely distributed box jellyfish found in the mediterranean and in the tropical waters of the caribbean sea. its venom is a complex mixture of biologically active compounds that are used to catch prey. in order to evaluate the activity of the neurotoxins in the venom, bioassays were carried out using the marine crab ocypode quadrata. the proteins with neurotoxic effect were partially purified using low-pressure liquid chromatography techniques. gel filtration (sephadex ...201727815048
conservatism and variability of gene expression profiles among homeologous transcription factors in xenopus laevis.xenopus laevis has an allotetraploid genome of 3.1gb, in contrast to the diploid genome of a closely related species, xenopus tropicalis. here, we identified 412 genes (189 homeolog pairs, one homeologous gene cluster pair, and 28 singletons) encoding transcription factors (tfs) in the x. laevis genome by comparing them with their orthologs from x. tropicalis. those genes include the homeobox gene family (mix/bix, lhx, nkx, paired, pou, and vent), sox, fox, pax, dmrt, hes, gata, t-box, and some ...201727810169
the antidepressant bupropion is a negative allosteric modulator of serotonin type 3a receptors.the fda-approved antidepressant and smoking cessation drug bupropion is known to inhibit dopamine and norepinephrine reuptake transporters, as well as nicotinic acetylcholine receptors (nachrs) which are cation-conducting members of the cys-loop superfamily of ion channels, and more broadly pentameric ligand-gated ion channels (plgics). in the present study, we examined the ability of bupropion and its primary metabolite hydroxybupropion to block the function of cation-selective serotonin type 3 ...201727671323
a sister chromatid cohesion assay using xenopus egg extracts.cell-free extracts made from xenopus laevis eggs enable us to recapitulate many chromosomal events associated with cell cycle progression in a test tube. when sperm chromatin is incubated with these extracts, it is first duplicated within an assembled nucleus, and is then transformed into mitotic chromosomes, in each of which sister chromatids are juxtaposed with each other in a cohesin-dependent manner. here we describe our protocols for assembling duplicated chromosomes using egg extracts, alo ...201727797070
characterization of human short chain dehydrogenase/reductase sdr16c family members related to retinol dehydrogenase 10.all-trans-retinoic acid (ra) is a bioactive derivative of vitamin a that serves as an activating ligand for nuclear transcription factors, retinoic acid receptors. ra biosynthesis is initiated by the enzymes that oxidize retinol to retinaldehyde. it is well established that retinol dehydrogenase 10 (rdh10, sdr16c4), which belongs to the 16c family of the short chain dehydrogenase/reductase (sdr) superfamily of proteins, is the major enzyme responsible for the oxidation of retinol to retinaldehyd ...201727793605
methylmercury induces an initial increase in gaba-evoked currents in xenopus oocytes expressing α1 and α6 subunit-containing gabaa receptors.early onset effects of methylmercury (mehg) on recombinant α1β2γ2s or α6β2γ2s subunit-containing gabaa receptors were examined. these are two of the most prevalent receptor types found in cerebellum-a consistent target of mehg-induced neurotoxicity. heterologously expressed receptors were used in order to: (1) isolate receptor-mediated events from extraneous effects of mehg due to stimulation of the receptor secondary to increased release of gaba seen with mehg in neurons in situ and (2) limit t ...201727720918
tissue-specific selenium accumulation and toxicity in adult female xenopus laevis chronically exposed to elevated dietary selenomethionine.selenium (se) is a developmental toxicant that is also capable of altering the bioenergetic and endocrine status of adult fish. to date, aquatic ecotoxicological research has predominantly focused on the toxic effects of se in fish, and minimal information has been published related to amphibians. the objective of the present study was to investigate the potential toxicity associated with chronically elevated dietary se consumption in adult female amphibians utilizing the model species xenopus l ...201727666932
developmental changes in head movement kinematics during swimming in xenopus laevis tadpoles.during the post-embryonic developmental growth of animals, a number of physiological parameters such as locomotor performance, dynamics and behavioural repertoire are adjusted to match the requirements determined by changes in body size, proportions and shape. moreover, changes in movement parameters also cause changes in the dynamics of self-generated sensory stimuli, to which motion-detecting sensory systems have to adapt. here, we examined head movements and swimming kinematics of xenopus lae ...201727811303
characterization of tilapia (oreochromis niloticus) aldehyde reductase (akr1a1) gene, promoter and expression pattern in benzo-a-pyrene exposed fish.this study planned to isolation and characterization of akr1a1 cdna from bap injected nile tilapia (oreochromis niloticus), comparison of its characteristic structures with those of other species, characterization of akr1a1 gene and promoter, and investigation of akr1a1 mrna expression in various organs of bap injected tilapia. the cdna was 1172 bp long which includes an open reading frame of 975 bp encoding a 324 amino acids protein and a stop codon. the sequence showed 3' and 5' non-coding reg ...201727800707
characterization of cadmium chloride-induced bip accumulation in xenopus laevis a6 kidney epithelial cells.endoplasmic reticulum (er) stress can result in the accumulation of unfolded/misfolded protein in the er lumen, which can trigger the unfolded protein response (upr) resulting in the activation of various genes including immunoglobulin-binding protein (bip; also known as glucose-regulated protein 78 or hspa5). bip, an er heat shock protein 70 (hsp70) family member, binds to unfolded protein, inhibits their aggregation and re-folds them in an atp-dependent manner. while cadmium, an environmental ...201727746171
rhythm generation, coordination, and initiation in the vocal pathways of male african clawed frogs.central pattern generators (cpgs) in the brain stem are considered to underlie vocalizations in many vertebrate species, but the detailed mechanisms underlying how motor rhythms are generated, coordinated, and initiated remain unclear. we addressed these issues using isolated brain preparations of xenopus laevis from which fictive vocalizations can be elicited. advertisement calls of male x. laevis that consist of fast and slow trills are generated by vocal cpgs contained in the brain stem. brai ...201727760822
the extracellular c-loop domain plays an important role in the cell adhesion function of aquaporin 0.although aquaporin 0 (aqp0) is a member of the aqp family, it has limited water permeability compared with other members. aqp0 may also have cell adhesion-related functions, but the evidence is still limited. here, we studied the relationship of aqp0 to cell adhesion and determined the region required for cell adhesion.201727754715
identification and comparative analyses of siamois cluster genes in xenopus laevis and tropicalis.two siamois-related homeobox genes siamois (sia1) and twin (sia2), have been reported in xenopus laevis. these genes are expressed in the blastula chordin- and noggin-expressing (bcne) center and the nieuwkoop center, and have complete secondary axis-inducing activity when over-expressed on the ventral side of the embryo. using whole genome sequences of x. tropicalis and x. laevis, we identified two additional siamois-related genes, which are tandemly duplicated near sia1 and sia2 to form the si ...201727522305
from the cover: functional analysis reveals glutamate and gamma-aminobutyric acid-gated chloride channels as targets of avermectins in the carmine spider mite.the carmine spider mite, tetranychus cinnabarinus (boisduval), is one of the most important pests in the agricultural industry. avermectins, as efficient acaricides, have been extensively used in the mite control. the targets for avermectins in mammals, nematodes, and insects had already been identified as γ-aminobutyric acid and glutamate chloride channels. however, the targets for avermectins in mites are still not clear. here, we report the cloning of 5 full-length glutamate-gated chloride ch ...201727742867
primordial germ cell isolation from xenopus laevis embryos.primordial germ cells (pgcs) are the precursors to the gametes and have the unique ability to retain full developmental potential. however, the mechanism(s) and gene-network(s) necessary for their proper specification and development are poorly understood. this is due, in part, to the challenges that must be overcome in order to identify and isolate pgcs during critical stages of development. two distinct mechanisms have been characterized to specify the germ cell lineage in vertebrates: inducti ...201727734352
genome organization of the vg1 and nodal3 gene clusters in the allotetraploid frog xenopus laevis.extracellular factors belonging to the tgf-β family play pivotal roles in the formation and patterning of germ layers during early xenopus embryogenesis. here, we show that the vg1 and nodal3 genes of xenopus laevis are present in gene clusters on chromosomes xla1l and xla3l, respectively, and that both gene clusters have been completely lost from the syntenic s chromosome regions. the presence of gene clusters and chromosome-specific gene loss were confirmed by cdna fish analyses. sequence and ...201727720224
the african clawed frog xenopus laevis: a model organism to study regeneration of the central nervous system.while an injury to the central nervous system (cns) in humans and mammals is irreversible, amphibians and teleost fish have the capacity to fully regenerate after severe injury to the cns. xenopus laevis has a high potential to regenerate the brain and spinal cord during larval stages (47-54), and loses this capacity during metamorphosis. the optic nerve has the capacity to regenerate throughout the frog's lifespan. here, we review cns regeneration in frogs, with a focus in x. laevis, but also p ...201727693567
genomic organization and modulation of gene expression of the tgf-β and fgf pathways in the allotetraploid frog xenopus laevis.inductive interactions mediated by the tgf-β and fgf-mapk pathways are essential for specification of the germ layers and embryonic body axes during early vertebrate embryogenesis. tgf-β and fgf ligands signal through receptor ser/thr and tyr kinases, respectively, and these signaling pathways cross-talk to regulate transcription and cell behavior. the allotetraploid xenopus laevis and its ancestral diploid xenopus tropicalis are versatile model organisms with which to study the inductive intera ...201727692744
effects of freezing and activation on membrane quality and dna damage in xenopus tropicalis and xenopus laevis spermatozoa.there is growing concern over the effect of sperm cryopreservation on dna integrity and the subsequent development of offspring generated from this cryopreserved material. in the present study, membrane integrity and dna stability of xenopus laevis and xenopus tropicalis spermatozoa were evaluated in response to cryopreservation with or without activation, a process that happens upon exposure to water to spermatozoa of some aquatic species. a dye exclusion assay revealed that sperm plasma membra ...201727692061
electrophysiological actions of synthetic cathinones on monoamine transporters.products containing psychoactive synthetic cathinones, such as mephedrone and 3,4-methylenedioxypyrovalerone (mdpv) are prevalent in our society. synthetic cathinones are structurally similar to methamphetamine, and numerous synthetics have biological activity at dopamine, serotonin, and norepinephrine transporters. importantly, monoamine transporters co-transport sodium ions along with their substrate, and movement of substrates and ions through the transporter can generate measurable ionic cur ...201727677783
an implantable eng detector with in-system velocity selective recording (vsr) capability.detection and classification of electroneurogram (eng) signals in the peripheral nervous system can be achieved by velocity selective recording (vsr) using multi-electrode arrays. this paper describes an implantable vsr-based eng recording system representing a significant development in the field since it is the first system of its type that can record naturally evoked eng and be interfaced wirelessly using a low data rate transcutaneous link. the system consists of two cmos asics one of which ...201727638107
the sox transcriptional factors: functions during intestinal development in vertebrates.the intestine has long been studied as a model for adult stem cells due to the life-long self-renewal of the intestinal epithelium through the proliferation of the adult intestinal stem cells. recent evidence suggests that the formation of adult intestinal stem cells in mammals takes place during the thyroid hormone-dependent neonatal period, also known as postembryonic development, which resembles intestinal remodeling during frog metamorphosis. studies on the metamorphosis in xenopus laevis ha ...201727567710
insulin-like growth factor 1 regulation of proliferation and differentiation of xenopus laevis myogenic cells in vitro.to understand the mechanism of muscle remodeling during xenopus laevis metamorphosis, we examined the in vitro effect of insulin-like growth factor 1 (igf-1) on growth and differentiation of three different-fate myogenic cell populations: tadpole tail, tadpole dorsal, and young adult leg muscle. igf-1 promoted growth and differentiation of both tail and leg myogenic cells only under conditions where these cells could proliferate. inhibition of cell proliferation by dna synthesis inhibitor cytosi ...201727699652
robust identification of ptbp1-dependent splicing events by a junction-centric approach in xenopus laevis.regulation of alternative splicing is an important process for cell differentiation and development. down-regulation of ptbp1, a regulatory rna-binding protein, leads to developmental skin defects in xenopus laevis. to identify ptbp1-dependent splicing events potentially related to the phenotype, we conducted rnaseq experiments following ptbp1 depletion. we systematically compared exon-centric and junction-centric approaches to detect differential splicing events. we showed that the junction-cen ...201727546377
gene expression analysis of developing cell groups in the pretectal region of xenopus laevis.our previous analysis of progenitor domains in the pretectum of xenopus revealed three molecularly distinct anteroposterior subdivisions, identified as precommissural (pcp), juxtacommissural (jcp), and commissural (cop) histogenetic domains (morona et al. [2011] j comp neurol 519:1024-1050). here we analyzed at later developmental stages the nuclei derived from these areas, attending to their gene expression patterns and histogenesis. transcription-factor gene markers were used to selectively ma ...201727539385
genes coding for cyclin-dependent kinase inhibitors are fragile in xenopus.cell proliferation is strictly regulated by the dosage balance among cell-cycle regulators such as cdk/cyclin complexes and cdk-inhibitors. even in the allotetraploid genome of xenopus laevis, the dosage balance must be maintained for animals to stay alive, and the duplicated homeologous genes seem to have gradually changed, through evolution, resulting in the best genes for them to thrive. in the xenopus laevis genome, while homeologous gene pairs of cdks are fundamentally maintained and a few ...201727393661
expression pattern and pharmacological characterisation of two novel alternative splice variants of the glutamate-gated chloride channel in the small brown planthopper laodelphax striatellus.glutamate-gated chloride channels (glucl) mediate fast inhibitory neurotransmission in invertebrate nervous systems. although only one glucl gene was presented in insects, it showed diverse alternative splicing that was speculated could affect channel function and pharmacology.201727302648
in vitro models of cranial neural crest development toward toxicity tests: frog, mouse, and human.during craniofacial development, cranial neural crest (nc)-derived mesenchymal cells migrate to pharyngeal arches and contribute extensively to neurons, schwann cells, smooth muscle cells, osteoblasts, chondrocytes, and odontoblasts, forming maxillofacial structures. in vitro models using model organism cells, such as african clawed frog (xenopus laevis) and mouse (mus musculus), were developed to understand cellular and molecular mechanisms of cranial nc development. recent studies using human ...201727299949
sex chromosome differentiation and the w- and z-specific loci in xenopus laevis.genetic sex-determining systems in vertebrates include two basic types of heterogamety; xx (female)/xy (male) and zz (male)/zw (female) types. the african clawed frog xenopus laevis has a zz/zw-type sex-determining system. in this species, we previously identified a w-specific sex (female)-determining gene dmw, and specified w and z chromosomes, which could be morphologically indistinguishable (homomorphic). in addition to dmw, we most recently discovered two genes, named scanw and ccdc69w, and ...201727297884
translational profiling of retinal ganglion cell optic nerve regeneration in xenopus laevis.unlike adult mammals, adult frogs regrow their optic nerve following a crush injury, making xenopus laevis a compelling model for studying the molecular mechanisms that underlie neuronal regeneration. using translational ribosome affinity purification (trap), a method to isolate ribosome-associated mrnas from a target cell population, we have generated a transcriptional profile by rna-seq for retinal ganglion cells (rgc) during the period of recovery following an optic nerve injury. based on bio ...201727471010
xenmine: a genomic interaction tool for the xenopus community.the xenopus community has embraced recent advances in sequencing technology, resulting in the accumulation of numerous rna-seq and chip-seq datasets. however, easily accessing and comparing datasets generated by multiple laboratories is challenging. thus, we have created a central space to view, search and analyze data, providing essential information on gene expression changes and regulatory elements present in the genome. xenmine (www.xenmine.org) is a user-friendly website containing publishe ...201727157655
pharmacological characterization of histamine-gated chloride channels from the housefly musca domestica.the biogenic amine histamine (ha) is not only the neurotransmitter of photoreceptors but also has important roles in mechanosensory reception, temperature preference, sleep and olfactory processing in insects. two cdnas (mdhcla and mdhclb) that encode ha-gated chloride channel subunits (mdhcla and mdhclb) were cloned from the housefly musca domestica. the crnas were injected into xenopus laevis oocytes to examine the functions and pharmacological characteristics of mdhcla and mdhclb channels usi ...201727139242
exploring the functions of nonclassical mhc class ib genes in xenopus laevis by the crispr/cas9 system.a large family of highly related and clustered xenopus nonclassical mhc class ib (xnc) genes influences xenopus laevis immunity and potentially other physiological functions. using rna interference (rnai) technology, we previously demonstrated that one of xnc genes, xnc10.1, is critical for the development and function of a specialized innate t (it) cell population. however, rnai limitation such as a variable and unstable degree of gene silencing in f0 and f1 generations is hampering a thorough ...201727318386
expanding the genetic toolkit in xenopus: approaches and opportunities for human disease modeling.the amphibian model xenopus, has been used extensively over the past century to study multiple aspects of cell and developmental biology. xenopus offers advantages of a non-mammalian system, including high fecundity, external development, and simple housing requirements, with additional advantages of large embryos, highly conserved developmental processes, and close evolutionary relationship to higher vertebrates. there are two main species of xenopus used in biomedical research, xenopus laevis ...201727109192
xenopus: leaping forward in kidney organogenesis.while kidney donations stagnate, the number of people in need of kidney transplants continues to grow. although transplanting culture-grown organs is years away, pursuing the engineering of the kidney de novo is a valid means of closing the gap between the supply and demand of kidneys for transplantation. the structural organization of a mouse kidney is similar to that of humans. therefore, mice have traditionally served as the primary model system for the study of kidney development. the mouse ...201727099217
evaluating the teratogenicity of the selective ß3-adrenoceptor agonist, cl 316.243 hydrate by employing fetax (frog embryo teratogenesis assay).in this study, the frog embryo teratogenesis assay (fetax - xenopus) technique was employed to evaluate the potential teratogenicity of the selective ß-adrenoceptor (ar) agonist, cl 316.243. in this context, cl 316.243 was applied to the south african clawed frog (xenopus laevis) embryos. the media containing the cl 316.24-exposed embryos were monitored and changed/replaced once every 24 hours. using fetax, we determined the minimum concentrations to inhibit growth (mcig) for cl 316.243. the 96- ...201727063898
evaluation of microtransplantation of rat brain neurolemma into xenopus laevis oocytes as a technique to study the effect of neurotoxicants on endogenous voltage-sensitive ion channels.microtransplantation of mammalian brain neurolemma into the plasma membrane of xenopus oocytes is used to study ion channels in their native form as they appear in the central nervous system. use of microtransplanted neurolemma is advantageous for various reasons: tissue can be obtained from various sources and at different developmental stages; ion channels and receptors are present in their native configuration in their proper lipid environment along with appropriate auxiliary subunits; allowi ...201727063102
xenopus genomic data and browser resources.the two species of xenopus most commonly used in biomedical research are the diploid xenopus (silurana) tropicalis and the tetraploid xenopus laevis. the x. tropicalis genome sequence has been available since 2010 and this year the x. laevis, genome from two distinct genetic backgrounds has been published. multiple genome assemblies available for both species and transcriptomic and epigenetic data sets are growing rapidly, all of which are available from a variety of web resources. this review d ...201727039265
generation of a xenopus laevis f1 albino j strain by genome editing and oocyte host-transfer.completion of the xenopus laevis genome sequence from inbred j strain animals has facilitated the generation of germline mutant x. laevis using targeted genome editing. in the last few years, numerous reports have demonstrated that talens are able to induce mutations in f0 xenopus embryos, but none has demonstrated germline transmission of such mutations in x. laevis. in this report we used the oocyte host-transfer method to generate mutations in both tyrosinase homeologs and found highly-penetr ...201726993591
genome-wide analysis of dorsal and ventral transcriptomes of the xenopus laevis gastrula.rna sequencing has allowed high-throughput screening of differential gene expression in many tissues and organisms. xenopus laevis is a classical embryological and cell-free extract model system, but its genomic sequence had been lacking due to difficulties arising from allotetraploidy. there is currently much excitement surrounding the release of the completed x. laevis genome (version 9.1) by the joint genome institute (jgi), which provides a platform for genome-wide studies. here we present a ...201727016259
extinction of an introduced warm-climate alien species, xenopus laevis, by extreme weather events.invasive, non-native species represent a major threat to biodiversity worldwide. the african amphibian xenopus laevis is widely regarded as an invasive species and a threat to local faunas. populations originating at the western cape, south africa, have been introduced on four continents, mostly in areas with a similar mediterranean climate. some introduced populations are also established in cooler environments where persistence for many decades suggests a capacity for long-term adaptation. in ...201726430383
invasive american bullfrogs and african clawed frogs in south america: high suitability of occurrence in biodiversity hotspots.fabiana g. barbosa, camila both, and miguel b. araújo (2017) invasion of protected areas by non-native species is currently one of the main threats to global biodiversity. using an ensemble of bioclimatic envelope models we quantify the degree of exposure of south american protected areas to invasion by two invasive amphibian species. we focus on protected areas that coincide with global biodiversity hotspots. the species modeled, lithobates catesbeianus and xenopus laevis, have been reported to ...201731966227
functional variants in hcn4 and cacna1h may contribute to genetic generalized epilepsy.genetic generalized epilepsy (gge) encompasses seizure disorders characterized by spike-and-wave discharges (swd) originating within thalamo-cortical circuits. hyperpolarization-activated (hcn) and t-type ca2+ channels are key modulators of rhythmic activity in these brain regions. here, we screened hcn4 and cacna1h genes for potentially contributory variants and provide their functional analysis.201729588962
molecular characterization and tissue expression profiles of prepro-vasoactive intestinal peptide in the chinese alligator (alligator sinensis) during the active and hibernating periods.the chinese alligator (alligator sinensis), a freshwater crocodilian endemic to china, is one of the most endangered crocodilian species; up to this date, very little is known about the endocrine regulation of its metabolic activities during different physiological states. in this study, we characterized the structure of the prepro-vasoactive intestinal peptide in chinese alligator (prepro-cavip) for the first time and examined its expression profiles in various tissues during the active and hib ...201729356375
biometric data assessment on xenopus laeis tadpoles. 201630701053
structural heterogeneity of cnga1 channels revealed by electrophysiology and single-molecule force spectroscopy.the determination at atomic resolution of the three-dimensional molecular structure of membrane proteins such as receptors and several ion channels has been a major breakthrough in structural biology. the molecular structure of several members of the superfamily of voltage-gated ionic channels such as k+ and na+ is now available. however, despite several attempts, the molecular structure at atomic resolution of the full cyclic nucleotide-gated (cng) ion channel, although a member of the same sup ...201631457189
a matter of the heart: the african clawed frog xenopus as a model for studying vertebrate cardiogenesis and congenital heart defects.the african clawed frog, xenopus, is a valuable non-mammalian model organism to investigate vertebrate heart development and to explore the underlying molecular mechanisms of human congenital heart defects (chds). in this review, we outline the similarities between xenopus and mammalian cardiogenesis, and provide an overview of well-studied cardiac genes in xenopus, which have been associated with congenital heart conditions. additionally, we highlight advantages of modeling candidate genes deri ...201629367567
in vitro model systems to investigate bile salt export pump (bsep) activity and drug interactions: a review.the bile salt export pump protein (bsep), expressed on the canalicular membranes of hepatocytes, is primarily responsible for the biliary excretion of bile salts. the inhibition of bsep transport activity can lead to an increase in intracellular bile salt levels and liver injury. this review discusses the various in vitro assays currently available for assessing the effect of drugs or other chemical entities to modulate bsep transport activity. bsep transporter assays use one of the following pl ...201626683212
quantitative analysis of filament branch orientation in listeria actin comet tails.several bacterial and viral pathogens hijack the host actin cytoskeleton machinery to facilitate spread and infection. in particular, listeria uses arp2/3-mediated actin filament nucleation at the bacterial surface to generate a branched network that will help propel the bacteria. however, the mechanism of force generation remains elusive due to the lack of high-resolution three-dimensional structural data on the spatial organization of the actin mother and daughter (i.e., branch) filaments with ...201626497103
effects of dietary exposure of polycyclic musk hhcb on the metamorphosis of xenopus laevis.the compound 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta-[γ]-2-benzopyrane (hhcb; galaxolide, chemical abstracts service number 1222-05-5) is a synthetic musk used extensively as a fragrance in many consumer products and classified as an emerging pollutant. the ecotoxicological information available for hhcb addresses exposure via water, but this compound is frequently adsorbed into particulate matter. the goal of the present study was to assess the effects of dietary exposure to seve ...201626472276
catalytic activity of human carbonic anhydrase isoform ix is displayed both extra- and intracellularly.most carbonic anhydrases catalyse the reversible conversion of carbon dioxide to protons and bicarbonate, either as soluble cytosolic enzymes, in or at intracellular organelles, or at the extracellular face of the cell membrane as membrane-anchored proteins. carbonic anhydrase isoform ix (ca ix), a membrane-bound enzyme with catalytic activity at the extracellular membrane surface, has come to prominence in recent years because of its association with hypoxic tissue, particularly tumours, often ...201626470855
bumetanide derivatives aqb007 and aqb011 selectively block the aquaporin-1 ion channel conductance and slow cancer cell migration.aquaporins (aqps) in the major intrinsic family of proteins mediate fluxes of water and other small solutes across cell membranes. aqp1 is a water channel, and under permissive conditions, a nonselective cation channel gated by cgmp. in addition to mediating fluid transport, aqp1 expression facilitates rapid cell migration in cell types including colon cancers and glioblastoma. work here defines new pharmacological derivatives of bumetanide that selectively inhibit the ion channel, but not the w ...201626467039
phosphorylation of rs1 (rsc1a1) steers inhibition of different exocytotic pathways for glucose transporter sglt1 and nucleoside transporter cnt1, and an rs1-derived peptide inhibits glucose absorption.cellular uptake adapts rapidly to physiologic demands by changing transporter abundance in the plasma membrane. the human gene rsc1a1 codes for a 67-kda protein named rs1 that has been shown to induce downregulation of the sodium-d-glucose cotransporter 1 (sglt1) and of the concentrative nucleoside transporter 1 (cnt1) in the plasma membrane by blocking exocytosis at the golgi. injecting rs1 fragments into xenopus laevis oocytes expressing sglt1 or cnt1 and measuring the expressed uptake of α-me ...201626464324
a curcumin analog, go-y078, effectively inhibits angiogenesis through actin disorganization.the inhibition of angiogenesis is a theoretically ideal chemotherapy for cancer, but there remains room for improvement. most inhibitors of angiogenesis approved to date target vascular endothelial growth factors (vegfs); however, vegfs are only one of the many classes of participant in tumor angiogenesis. because tumor angiogenesis is orchestrated by many components, including growth factors, signal transducers, and effectors, its regulation exhibits redundancy. curcumin can associate with many ...201626459768
complicated behavior of g-quadruplexes and evaluating g-quadruplexes' ligands in various systems mimicking cellular circumstance.environments surrounding g-rich sequences remarkably affect the conformations of these structures. a proper evaluation system mimicking the crowded environment in a cell with macromolecules should be developed to perform structural and functional studies on g-quadruplexes. in this study, the topology and stability of a g-quadruplex formed by human telomeric repeat sequences were investigated in a macromolecule-crowded environment created by polyethylene glycol 200 (peg200), tumor cell extract, a ...201628955851
proteomics of xenopus development.modern mass spectrometry-based methods provide an exciting opportunity to characterize protein expression in the developing embryo. we have employed an isotopic labeling technology to quantify the expression dynamics of nearly 6000 proteins across six stages of development in xenopus laevis from the single stage zygote through the mid-blastula transition and the onset of organogenesis. approximately 40% of the proteins show significant changes in expression across the development stages. the exp ...201626396253
amphibian macrophage development and antiviral defenses.macrophage lineage cells represent the cornerstone of vertebrate physiology and immune defenses. in turn, comparative studies using non-mammalian animal models have revealed that evolutionarily distinct species have adopted diverse molecular and physiological strategies for controlling macrophage development and functions. notably, amphibian species present a rich array of physiological and environmental adaptations, not to mention the peculiarity of metamorphosis from larval to adult stages of ...201626705159
electrophysiological characterization of na,k-atpases expressed in xenopus laevis oocytes using two-electrode voltage clamping.the transport of three na(+) per two k(+) means that the na,k-atpase is electrogenic, and though the currents generated by the ion pump are small compared to ion channel currents, they can be measured using electrophysiology, both steady-state pumping and individual steps in the transport cycle. various electrophysiological techniques have been used to study the endogenous pumps of the squid giant axon and of cardiac myocytes from for example rabbits. here, we describe the characterization of he ...201626695042
identification of methylated deoxyadenosines in vertebrates reveals diversity in dna modifications.methylation of cytosine deoxynucleotides generates 5-methylcytosine (m(5)dc), a well-established epigenetic mark. however, in higher eukaryotes much less is known about modifications affecting other deoxynucleotides. here, we report the detection of n(6)-methyldeoxyadenosine (m(6)da) in vertebrate dna, specifically in xenopus laevis but also in other species including mouse and human. our methylome analysis reveals that m(6)da is widely distributed across the eukaryotic genome and is present in ...201626689968
single-cell mass spectrometry for discovery proteomics: quantifying translational cell heterogeneity in the 16-cell frog (xenopus) embryo.we advance mass spectrometry from a cell population-averaging tool to one capable of quantifying the expression of diverse proteins in single embryonic cells. our instrument combines capillary electrophoresis (ce), electrospray ionization, and a tribrid ultrahigh-resolution mass spectrometer (hrms) to enable untargeted (discovery) proteomics with ca. 25 amol lower limit of detection. ce-μesi-hrms enabled the identification of 500-800 nonredundant protein groups by measuring 20 ng, or <0.2% of th ...201626756663
homology-driven assembly of non-redundant protein sequence sets (nomess) for mass spectrometry.to enable mass spectrometry (ms)-based proteomic studies with poorly characterized organisms, we developed a computational workflow for the homology-driven assembly of a non-redundant reference sequence dataset. in the automated pipeline, translated dna sequences (e.g. ests, rna deep-sequencing data) are aligned to those of a closely related and fully sequenced organism. representative sequences are derived from each cluster and joined, resulting in a non-redundant reference set representing the ...201626743511
atrazine and malathion shorten the maturation process of xenopus laevis oocytes and have an adverse effect on early embryo development.the use of pesticides has a negative impact on the environment. amphibians have long been regarded as indicator species to pollutants due to their permeable skin and sensitivity to the environment. studies have shown that population declines of some amphibians are directly linked with exposure to agricultural contaminants. in the past, much of the studies have focused on the toxic effect of contaminants on larvae (tadpoles), juvenile and adult frogs. however, due to the nature of their life cycl ...201626686575
nearly 1000 protein identifications from 50 ng of xenopus laevis zygote homogenate using online sample preparation on a strong cation exchange monolith based microreactor coupled with capillary zone electrophoresis.a sulfonate-silica hybrid strong cation exchange monolith microreactor was synthesized and coupled to a linear polyacrylamide coated capillary for online sample preparation and capillary zone electrophoresis-tandem mass spectrometry (cze-ms/ms) bottom-up proteomic analysis. the protein sample was loaded onto the microreactor in an acidic buffer. after online reduction, alkylation, and digestion with trypsin, the digests were eluted with 200 mm ammonium bicarbonate at ph 8.2 for cze-ms/ms analysi ...201626670623
expression pattern of bcar3, a downstream target of gata2, and its binding partner, bcar1, during xenopus development.primitive hematopoiesis generates red blood cells that deliver oxygen to the developing embryo. mesodermal cells commit to a primitive blood cell fate during gastrulation and, in order to do so the mesoderm must receive non-cell autonomous signals transmitted from other germ layers. in xenopus, the transcription factor gata2 functions in ectodermal cells to generate or transmit the non-cell autonomous signals. here we have identified breast cancer antiestrogen resistance 3 (bcar3) as a gene that ...201626631802
differential thyroid hormone sensitivity of fast cycling progenitors in the neurogenic niches of tadpoles and juvenile frogs.adult neurogenesis occurs in neural stem cell (nsc) niches where slow cycling stem cells give rise to faster cycling progenitors. in the adult mouse nsc niche thyroid hormone, t3, and its receptor trα act as a neurogenic switch promoting progenitor cell cycle completion and neuronal differentiation. little is known about whether and how t3 controls proliferation of differentially cycling cells during xenopus neurogenesis. to address this question, we first used sox3 as a marker of stem cell and ...201626628040
sex differences and endocrine regulation of auditory-evoked, neural responses in african clawed frogs (xenopus).mating depends on the accurate detection of signals that convey species identity and reproductive state. in african clawed frogs, xenopus, this information is conveyed by vocal signals that differ in temporal patterns and spectral features between sexes and across species. we characterized spectral sensitivity using auditory-evoked potentials (aeps), commonly known as the auditory brainstem response, in males and females of four xenopus species. in female x. amieti, x. petersii, and x. laevis, p ...201626572136
identification of an inhibitory alcohol binding site in gabaa ρ1 receptors.alcohols inhibit γ-aminobutyric acid type a ρ1 receptor function. after introducing mutations in several positions of the second transmembrane helix in ρ1, we studied the effects of ethanol and hexanol on gaba responses using two-electrode voltage clamp electrophysiology in xenopus laevis oocytes. the 6' mutations produced the following effects on ethanol and hexanol responses: small increase or no change (t6'm), increased inhibition (t6'v), and small potentiation (t6'y and t6'f). the 5' mutatio ...201626571107
quantification of x. laevis vitellogenin by liquid chromatography tandem mass spectrometry.over the last several decades, there has been an increase in public awareness and regulatory activity in regard to the presence of emerging contaminants in the environment that may have the potential to interact with the endocrine system of exposed wildlife. alterations in vitellogenin (vtg), a high density yolk precursor protein, can indicate endocrine activity in oviparous species, including many fish and amphibians. while various methodologies and experiments have been performed to characteri ...201626562177
distinctive effects of nicotinic receptor intracellular-loop mutations associated with nocturnal frontal lobe epilepsy.previously characterized nicotinic acetylcholine receptor (nachr) autosomal dominant nocturnal frontal lobe epilepsy (adnfle)-associated mutations are found in α2, α4 and β2 subunit transmembrane (tm) domains. they predominantly increase ach potency and, for β2-subunit mutants, increase macroscopic currents. two recently-identified mutations, α4(r336h) and β2(v337g), located in the intracellular cytoplasmic loop (c2) have been associated with non-familial nfle. effects of these mutations on α4β2 ...201626561946
alternative splice variant of the thiazide-sensitive nacl cotransporter: a novel player in renal salt handling.the thiazide-sensitive nacl cotransporter (ncc) is an important pharmacological target in the treatment of hypertension. the human slc12a3 gene, encoding ncc, gives rise to three isoforms. only the third isoform has been extensively investigated. the aim of the present study was, therefore, to establish the abundance and localization of the almost identical isoforms 1 and 2 (ncc1/2) in the human kidney and to determine their functional properties and regulation in physiological conditions. immun ...201626561651
identification of micrornas and microrna targets in xenopus gastrulae: the role of mir-26 in the regulation of smad1.micrornas (mirnas) are known to play diverse roles in the regulation of vertebrate development. to investigate mirna-target mrna relationships in embryonic development, we have carried out small-rna sequencing to identify mirnas expressed in the early gastrula of xenopus laevis. we identify a total of 180 mirnas, and we have identified the locations of the mirna precursor sequences in the x. laevis genome. of these mirnas, 141 represent mirs previously identified in xenopus tropicalis. alignment ...201626548531
using in vivo imaging to measure rna mobility in xenopus laevis oocytes.rna localization in the xenopus oocyte is responsible for the establishment of polarity during oogenesis as well as the specification of germ layers during embryogenesis. however, the inability to monitor mrna localization in live vertebrate oocytes has posed a major barrier to understanding the mechanisms driving directional transport. here we describe a method for imaging ms2 tagged rna in live xenopus oocytes to study the dynamics of rna localization. we also focus on methods for implementing ...201626546269
sex-specific response to hypoxia in a reduced brainstem preparation from xenopus laevis.respiratory reflexes and tolerance to hypoxia show significant sexual dimorphism. however, the data supporting this notion originates exclusively from mammals. to determine whether this concept is limited to this group of vertebrates, we examined the sex-specific response to acute hypoxia in an adult reduced brainstem preparation from xenopus laevis. within the first 5min of exposure to hypoxic acsf (98% n2/2% co2), recordings of respiratory-related activity show a stronger increase in fictive b ...201626528898
the prototoxin lypd6b modulates heteromeric α3β4-containing nicotinic acetylcholine receptors, but not α7 homomers.prototoxins are a diverse family of membrane-tethered molecules expressed in the nervous system that modulate nicotinic cholinergic signaling, but their functions and specificity have yet to be completely explored. we tested the selectivity and efficacy of leukocyte antigen, plaur (plasminogen activator, urokinase receptor) domain-containing (lypd)-6b on α3β4-, α3α5β4-, and α7-containing nicotinic acetylcholine receptors (nachrs). to constrain stoichiometry, fusion proteins encoding concatemers ...201626586467
effect of n-glycosylation on the transport activity of the peptide transporter pept1.the intestinal peptide transporter pept1 provides bulk quantities of amino acids to epithelial cells. pept1 is a high-capacity and low-affinity solute carrier of the slc15 family found in apical membranes of enterocytes in small intestine and distal colon. surprisingly, murine pept1 (mpept1) has an apparent molecular mass of ∼95 kda in the small intestine but ∼105 kda in the large intestine. here we describe studies on mpept1 protein glycosylation and how glycans affect transport function. putat ...201626585416
monitoring smad activity in vivo using the xenopus model system.the embryo of the african clawed frog xenopus laevis plays a central role in the field of cell and developmental biology. one of the strengths of xenopus as model system lies in the high degree of conservation between amphibians and mammals in the molecular mechanisms controlling tissue patterning and differentiation. as such, many signaling cascades were first investigated in frog embryos and then confirmed in mouse and/or human cells. the tgf-β signaling cascade greatly benefited from this mod ...201626520129
mapping neurogenesis onset in the optic tectum of xenopus laevis.neural progenitor cells have a central role in the development and evolution of the vertebrate brain. during early brain development, neural progenitors first expand their numbers through repeated proliferative divisions and then begin to exhibit neurogenic divisions. the transparent and experimentally accessible optic tectum of xenopus laevis is an excellent model system for the study of the cell biology of neurogenesis, but the precise spatial and temporal relationship between proliferative an ...201627012549
neural regeneration dynamics of xenopus laevis olfactory epithelium after zinc sulfate-induced damage.neural stem cells (nscs) of the olfactory epithelium (oe) are responsible for tissue maintenance and the neural regeneration after severe damage of the tissue. in the normal oe, nscs are located in the basal layer, olfactory receptor neurons (orns) mainly in the middle layer, and sustentacular (sus) cells in the most apical olfactory layer. in this work, we induced severe damage of the oe through treatment with a zinc sulfate (znso4) solution directly in the medium, which resulted in the loss of ...201627012180
single-cell mass spectrometry with multi-solvent extraction identifies metabolic differences between left and right blastomeres in the 8-cell frog (xenopus) embryo.single-cell metabolic mass spectrometry enables the discovery (untargeted) analysis of small molecules in individual cells. using single-cell capillary electrophoresis high-resolution mass spectrometry (ce-hrms), we recently uncovered small-molecule differences between embryonic cells located along the animal-vegetal and dorsal-ventral axes of the 16-cell frog (xenopus laevis) embryo, raising the question whether metabolic cell heterogeneity also exists along the left-right body axis. to address ...201627004603
an early developmental vertebrate model for nanomaterial safety: bridging cell-based and mammalian toxicity assessment.with the rise in production of nanoparticles (nps) for an ever-increasing number of applications, there is an urgent need to efficiently assess their potential toxicity. we propose a np hazard assessment protocol that combines mammalian cytotoxicity data with embryonic vertebrate abnormality scoring to determine an overall toxicity index.201627003295
expressional characterization of mrna (guanine-7) methyltransferase (rnmt) during early development of xenopus laevis.methylation of the guanosine cap structure at the 5' end of mrna is essential for efficient translation of all eukaryotic cellular mrnas, gene expression and cell viability and promotes transcription, splicing, polyadenylation and nuclear export of mrna. in the current study, we present the spatial expression pattern of the xenopus laevis rnmt homologue. a high percentage of protein sequence similarity, especially within the methyltransferase domain, as well as an increased expression in the cel ...201627002806
reappraising the effects of artemisinin on the atpase activity of pfatp6 and serca1a e255l expressed in xenopus laevis oocytes. 201626733217
assessing differences in toxicity and teratogenicity of three phthalates, diethyl phthalate, di-n-propyl phthalate, and di-n-butyl phthalate, using xenopus laevis embryos.phthalates, compounds used to add flexibility to plastics, are ubiquitous in the environment. in particular, the diethyl (dep), di-n-propyl (dnpp), and di-n-butyl (dbp) phthalates were found to exert detrimental effects in both mammalian and non-mammalian studies, with toxic effects varying according to alkyl chain length. embryos of xenopus laevis, the african clawed frog, have been used to assess toxicity and teratogenicity of several compounds and serves as a model for assessing adverse and t ...201626730679
hydrophobic interactions between the s5 segment and the pore helix stabilizes the closed state of slo2.1 potassium channels.under normal physiological conditions, slo2.1k(+) channels are in a closed state unless activated by an elevation in [na(+)]i. fenamates such as niflumic acid also activate slo2.1. previous studies suggest that activation of slo2.1 channels is mediated by a conformational change in the selectivity filter, and not a widening of the aperture formed by the s6 segment bundle crossing as occurs in voltage-gated k(+) channels. it is unclear how binding of na(+) or fenamates is allosterically linked to ...201626724206
identifying domains of efhc1 involved in ciliary localization, ciliogenesis, and the regulation of wnt signaling.efhc1 encodes a ciliary protein that has been linked to juvenile myoclonic epilepsy. in ectodermal explants, derived from xenopus laevis embryos, the morpholino-mediated down-regulation of efhc1b inhibited multiciliated cell formation. in those ciliated cells that did form, axoneme but not basal body formation was inhibited. efhc1b morphant embryos displayed defects in central nervous system (cns) and neural crest patterning that were rescued by a efhc1b-gfp chimera. efhc1b-gfp localized to cili ...201626783883
differential requirement of bone morphogenetic protein receptors ia (alk3) and ib (alk6) in early embryonic patterning and neural crest development.bone morphogenetic proteins regulate multiple processes in embryonic development, including early dorso-ventral patterning and neural crest development. bmps activate heteromeric receptor complexes consisting of type i and type ii receptor-serine/threonine kinases. bmp receptors ia and ib, also known as alk3 and alk6 respectively, are the most common type i receptors that likely mediate most bmp signaling events. since early expression patterns and functions in xenopus laevis development have no ...201626780949
lens regeneration from the cornea requires suppression of wnt/β-catenin signaling.the frog, xenopus laevis, possesses a high capacity to regenerate various larval tissues, including the lens, which is capable of complete regeneration from the cornea epithelium. however, the molecular signaling mechanisms of cornea-lens regeneration are not fully understood. previous work has implicated the involvement of the wnt signaling pathway, but molecular studies have been very limited. iris-derived lens regeneration in the newt (wolffian lens regeneration) has shown a necessity for act ...201626778749
using frogs faces to dissect the mechanisms underlying human orofacial defects.in this review i discuss how xenopus laevis is an effective model to dissect the mechanisms underlying orofacial defects. this species has been particularly useful in studying the understudied structures of the developing face including the embryonic mouth and primary palate. the embryonic mouth is the first opening between the foregut and the environment and is critical for adult mouth development. the final step in embryonic mouth formation is the perforation of a thin layer of tissue covering ...201626778163
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