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muscarinic modulation of the xenopus laevis tadpole spinal mechanosensory pathway.muscarinic acetylcholine receptors (machrs) mediate effects of acetylcholine (ach) in many systems, including those involved in spinal functions like locomotion. in xenopus laevis tadpoles at two days old, a model vertebrate for motor control research, we investigated the role of machrs in the skin mechanosensory pathway. we found that machr activation by carbachol did not affect the sensory rohon-beard neuron properties. however, carbachol could hyperpolarise sensory interneurons and decrease t ...201829601952
the effects of 4-hydroxybenzoic acid identified from bamboo (dendrocalamus asper) shoots on kv1.4 channel.bamboo shoot has been used as a treatment for epilepsy in traditional chinese medicine for generations to treat neuronal disorders such as convulsive, dizziness and headaches. 4-hydroxybenzoic acid (4-hba) is a non-flavonoid phenol found abundantly in dendrocalamus asper shoots (bamboo), fruits (strawberries and apples) and flowers. kv1.4 is a rapidly inactivating shaker-related member of the voltage-gated potassium channels with two inactivation mechanisms; the fast n-type and slow c-type. it p ...201829599640
frog's dcs have it all in one.although the amphibian xenopus laevis produces antibodies as diversified as those from mammals in the primary repertoire, antibody affinity maturation after immunization is relatively poor and has been associated with a poor b cell selection of aid-mediated hypermutations and lack of germinal centers in the spleen, the only secondary lymphoid organ of this amphibian. in this issue of the european journal of immunology, neely et al., [eur. j. immunol. 2018. 48: 430-440] have unveiled the role of ...201829522243
transcriptome analysis identifies genes involved in sex determination and development of xenopus laevis gonads.development of the gonads is a complex process, which starts with a period of undifferentiated, bipotential gonads. during this period the expression of sex-determining genes is initiated. sex determination is a process triggering differentiation of the gonads into the testis or ovary. sex determination period is followed by sexual differentiation, i.e. appearance of the first testis- and ovary-specific features. in xenopus laevis w-linked dm-domain gene (dm-w) had been described as a master det ...201829518581
c8orf46 homolog encodes a novel protein vexin that is required for neurogenesis in xenopus laevis.neural basic helix-loop helix (bhlh) transcription factors promote progenitor cell differentiation by activation of downstream target genes that coordinate neuronal differentiation. here we characterize a neural bhlh target gene in xenopus laevis, vexin (vxn; previously sbt1), that is homologous to human c8orf46 and is conserved across vertebrate species. c8orf46 has been implicated in cancer progression, but its function is unknown. vxn is transiently expressed in differentiating progenitors in ...201829518376
investigation of higher-order rna g-quadruplex structures in vitro and in living cells by 19f nmr spectroscopy.growing evidence indicates that rna g-quadruplexes have important roles in various processes such as transcription, translation, regulation of telomere length, and formation of telomeric heterochromatin. investigation of rna g-quadruplex structures associated with biological events is therefore essential to understanding the functions of these rna molecules. we recently demonstrated that the sensitivity and simplicity of 19f nmr can be used to directly observe higher-order telomeric g-quadruplex ...201829517770
occurrence of batrachochytrium dendrobatidis in anurans of the mediterranean region of baja california, méxico.chytridiomycosis is caused by the fungal pathogen batrachochytrium dendrobatidis (bd) and is regarded as one of the most significant threats to global amphibian populations. in méxico, bd was first reported in 2003 and has now been documented in 13 states. we visited 33 localities and swabbed 199 wild-caught anurans from 7 species (5 native, 2 exotic) across the mediterranean region of the state of baja california. using quantitative pcr, bd was detected in 94 individuals (47.2% of samples) at 2 ...201829516858
evaluation of the toxic effects of celecoxib on xenopus embryo development.celecoxib is a non-steroidal anti-inflammatory drug that selectively inhibits cyclooxygenase-2 and is prescribed for severe pain and inflammation. the excellent therapeutic effects of celecoxib mean that it is frequently used clinically, including for women of child-bearing age. however, the prenatal effects of this compound have not been studied extensively in vertebrates. the present study examined the developmental toxicity of celecoxib using a frog embryo teratogenic assay-xenopus (fetax). i ...201829505793
a novel rhodopsin phosphodiesterase from salpingoeca rosetta shows light-enhanced substrate affinity.it is since many years textbook knowledge that the concentration of the second messenger cgmp is regulated in animal rod and cone cells by type ii rhodopsins via a g-protein signaling cascade. microbial rhodopsins with enzymatic activity for regulation of cgmp concentration were only recently discovered: in 2014 light-activated guanylyl-cyclase opsins in fungi and in 2017 a novel rhodopsin phosphodiesterase (rhopde) in the protist salpingoeca rosetta (srrhopde). the light regulation of srrhopde, ...201829483295
effects of copper on the early development of xenopus laevis: the case of cuso4 and bordeaux mixture solutions. 201830701063
time-lapse imaging of cell death in cell culture and whole living organisms using turn-on deep-red fluorescent probes.cell death is a central process in developmental biology and also an important indicator of disease status and treatment efficacy. two related fluorescent probes are described that are molecular conjugates of one or two zinc dipicolylamine (zndpa) coordination complexes with an appended solvatochromic benzothiazolium squaraine dye. the probes were designed to target the anionic phospholipid, phosphatidylserine (ps), that is exposed on the surface of dead and dying cells. a series of spectrometri ...201830858977
the effect of an environmentally relevant phthalate mixture on primordial germ cells of xenopus laevis embryos. 201832550375
thyroid hormone levels and temperature during development alter thermal tolerance and energetics of xenopus laevis larvae.environmental variation induced by natural and anthropogenic processes including climate change may threaten species by causing environmental stress. anuran larvae experiencing environmental stress may display altered thyroid hormone (th) status with potential implications for physiological traits. therefore, any capacity to adapt to environmental changes through plastic responses provides a key to determining species vulnerability to environmental variation. we investigated whether developmenta ...201830464840
auditory perception exhibits sexual dimorphism and left telencephalic dominance in xenopus laevis.sex differences in both vocalization and auditory processing have been commonly found in vocal animals, although the underlying neural mechanisms associated with sexual dimorphism of auditory processing are not well understood. in this study we investigated whether auditory perception exhibits sexual dimorphism in xenopus laevis to do this we measured event-related potentials (erps) evoked by white noise (wn) and conspecific calls in the telencephalon, diencephalon and mesencephalon respectively ...201830509903
modeling underwater hearing and sound localization in the frog xenopus laevis.animals that are small compared to sound wavelengths face the challenge of localizing a sound source since the main cues to sound direction-interaural time differences (itd) and interaural level differences (ild)-both depend on size. remarkably, the majority of terrestrial vertebrates possess internally coupled ears (ice) with an air-filled cavity connecting the two eardrums and producing an inherently directional middle-ear system. underwater, longer wavelengths and faster sound-speed reduce bo ...201830522324
silurana chromosomal evolution: a new piece to the puzzle.the african clawed frogs of the subgenus silurana comprise both diploid and tetraploid species. the root of the polyploidization event leading to the extant xenopus calcaratus, x. mellotropicalis, and x. epitropicalis is not fully understood so far. in x. mellotropicalis, we previously proposed 2 evolutionary scenarios encompassing complete (scenario a) or incomplete (scenario b) translocation of a heterochromatic block from chromosome 9 to 2 in a diploid ancestor. to resolve this puzzle, we per ...201830537723
the interplay between voluntary food intake, dietary carbohydrate-lipid ratio and nutrient metabolism in an amphibian, (xenopus laevis).digestion of food and metabolism of frogs are little studied at the moment, and such processes could be very particular in the case of amphibians, given their ectothermic and carnivorous nature which may lead them to use nutrients through specific biochemical pathways. in the present study, 24 adult xenopus laevis (six replicates with two frogs per treatment) were randomly assigned to two diets with different carbohydrate:fat ratio (4.5:1 and 2.1:1), changing the dietary glucogenic and lipogenic ...201830532132
effects of quinine, quinidine and chloroquine on human muscle nicotinic acetylcholine receptors.the genus cinchona is known for a range of alkaloids, such as quinine, quinidine, cinchonine, and cinchonidine. cinchona bark has been used as an antimalarial agent for more than 400 years. quinine was first isolated in 1820 and is still acknowledged in the therapy of chloroquine-resistant falciparum malaria; in lower dosage quinine has been used as treatment for leg cramps since the 1940s. here we report the effects of the quinoline derivatives quinine, quinidine, and chloroquine on human adult ...201830515099
ventx1.1 as a direct repressor of early neural gene zic3 in xenopus laevis.from xenopus embryo studies, the bmp4/smad1-targeted gene circuit is a key signaling pathway for specifying the cell fate between the ectoderm and neuro-ectoderm as well as the ventral and dorsal mesoderm. in this context, several bmp4/smad1 target transcriptional factors have been identified as repressors of the neuro-ectoderm. however, none of these direct target transcription factors in this pathway, including gata1b, msx1 and ventx1.1 have yet been proven as direct repressors of early neuro- ...201830590909
development of a rapid functional assay that predicts glut1 disease severity.to examine the genotype to phenotype connection in glucose transporter type 1 (glut1) deficiency and whether a simple functional assay can predict disease outcome from genetic sequence alone.201830588498
xenopus hybrids provide insight into cell and organism size control.determining how size is controlled is a fundamental question in biology that is poorly understood at the organismal, cellular, and subcellular levels. the xenopus species, x. laevis and x. tropicalis differ in size at all three of these levels. despite these differences, fertilization of x. laevis eggs with x. tropicalis sperm gives rise to viable hybrid animals that are intermediate in size. we observed that although hybrid and x. laevis embryogenesis initiates from the same sized zygote and pr ...201830564147
alteration of the retinoid acid-cbp signaling pathway in neural crest induction contributes to enteric nervous system disorder.hirschsprung disease (hscr) and/or hypoganglionosis are common pediatric disorders that arise from developmental deficiencies of enteric neural crest cells (enccs). retinoid acid (ra) signaling has been shown to affect neural crest (nc) development. however, the mechanisms underlying ra deficiency-induced hscr or hypoganglionosis are not well-defined. in this report, we found that in hscr patient bowels, the ra nuclear receptor rarα and its interacting coregulator creb-binding protein (cbp) were ...201830560112
control of xenopus tadpole locomotion via selective expression of ih in excitatory interneurons.locomotion relies on the coordinated activity of rhythmic neurons in the hindbrain and spinal cord and depends critically on the intrinsic properties of excitatory interneurons. therefore, understanding how ion channels sculpt the properties of these interneurons, and the consequences for circuit function and behavior, is an important task. the hyperpolarization-activated cation current, ih, is known to play important roles in shaping neuronal properties and for rhythm generation in many neurona ...201830503615
genomic takeover by transposable elements in the strawberry poison frog.we sequenced the genome of the strawberry poison frog, oophaga pumilio, at a depth of 127.5× using variable insert size libraries. the total genome size is estimated to be 6.76 gb, of which 4.76 gb are from high copy number repetitive elements with low differentiation across copies. these repeats encompass dna transposons, rna transposons, and ltr retrotransposons, including at least 0.4 and 1.0 gb of mariner/tc1 and gypsy elements, respectively. expression data indicate high levels of gypsy and ...201830517748
novel findings on the impact of chytridiomycosis on the cardiac function of anurans: sensitive vs. tolerant species.understanding of the physiological effects of chytridiomycosis is crucial to worldwide amphibian conservation. therefore, we analyzed the cardiac function of two anuran species (xenopus laevis and physalaemus albonotatus) with different susceptibilities to infection by the causative agent of chytridiomycosis, batrachochytrium dendrobatidis (hereafter bd).201830425891
a taxonomically and geographically constrained information base limits non-native reptile and amphibian risk assessment: a systematic review.for many taxa, new records of non-native introductions globally occur at a near exponential rate. we undertook a systematic review of peer-reviewed publications on non-native herpetofauna, to assess the information base available for assessing risks of future invasions, resulting in 836 relevant papers. the taxonomic and geographic scope of the literature was also compared to a published database of all known invasions globally. we found 1,116 species of herpetofauna, 95% of which were present i ...201830425887
divergent subgenome evolution after allopolyploidization in african clawed frogs (xenopus).whole genome duplication (wgd), the doubling of the nuclear dna of a species, contributes to biological innovation by creating genetic redundancy. one mode of wgd is allopolyploidization, wherein each genome from two ancestral species becomes a 'subgenome' of a polyploid descendant species. the evolutionary trajectory of a duplicated gene that arises from wgd is influenced both by natural selection that may favour redundant, new or partitioned functions, and by gene silencing (pseudogenization). ...201830341989
automating event-detection of brain neuron synaptic activity and action potential firing in vivo using a random-access multiphoton laser scanning microscope for real-time analysis.determining how a neuron computes requires an understanding of the complex spatiotemporal relationship between its input (e.g. synaptic input as a result of external stimuli) and action potential output. recent advances in in vivo, laser-scanning multiphoton technology, known as random-access microscopy (ram), can capture this relationship by imaging fluorescent light, emitted from calcium-sensitive biosensors responding to synaptic and action potential firing in a neuron's full dendritic arbor ...201830440280
expression of hyaluronan synthases upregulated by thyroid hormone is involved in intestinal stem cell development during xenopus laevis metamorphosis.during amphibian intestinal remodeling, thyroid hormone (th) induces adult stem cells, which newly generate the absorptive epithelium analogous to the mammalian one. we have previously shown that hyaluronan (ha) is newly synthesized and plays an essential role in the development of the stem cells via its major receptor cd44 in the xenopus laevis intestine. we here focused on ha synthase (has) and examined how the expression of has family genes is regulated during natural and th-induced metamorph ...201830430240
the receptor-like pseudokinase ghr1 is required for stomatal closure.guard cells control the aperture of stomatal pores to balance photosynthetic carbon dioxide uptake with evaporative water loss. stomatal closure is triggered by several stimuli that initiate complex signaling networks to govern the activity of ion channels. activation of slow anion channel1 (slac1) is central to the process of stomatal closure and requires the leucine-rich repeat receptor-like kinase (lrr-rlk) guard cell hydrogen peroxide-resistant1 (ghr1), among other signaling components. here ...201830361234
development of an acute method to deliver transgenes into the brains of adult xenopus laevis.the central vocal pathway of the african clawed frog, xenopus laevis, is a powerful vertebrate model to understand mechanisms underlying central pattern generation. however, fast and efficient methods of introducing exogenous genes into the neurons of adult x. laevis are currently not available. here, we systematically tested methods of transgene delivery into adult x. laevis neurons. although successfully used for tadpole neurons for over a decade, electroporation was not efficient in transfect ...201830416430
atlht1 transporter can facilitate the uptake and translocation of a glycinergic-chlorantraniliprole conjugate in arabidopsis thaliana.understanding of the transporters involved in the uptake and translocation of agrochemicals in plants could provide an opportunity to guide pesticide to the site of insect feeding. the product of arabidopsis thaliana gene atlht1 makes a major contribution to the uptake into the roots of free amino acids and some of their derivatives. here, a chlorantraniliprole-glycine conjugate (cap-gly-1) was tested for its affinity to atlht1 both in planta and in vitro. seedlings deficient in atlht1 exhibited ...201830403859
transcriptome analysis of xenopus orofacial tissues deficient in retinoic acid receptor function.development of the face and mouth is orchestrated by a large number of transcription factors, signaling pathways and epigenetic regulators. while we know many of these regulators, our understanding of how they interact with each other and implement changes in gene expression during orofacial development is still in its infancy. therefore, this study focuses on uncovering potential cooperation between transcriptional regulators and one important signaling pathway, retinoic acid, during developmen ...201830390632
flow cytometric analysis of xenopus laevis and x. tropicalis blood cells using acridine orange.automated blood cell counters can distinguish cells based on their size and the presence or absence of a nucleus. however, most vertebrates have nucleated blood cells that cannot be counted automatically. we established an alternative automatic method for counting peripheral blood cells by staining cells with the fluorescent dye acridine orange (ao) and analysing cell populations using flow cytometry (fcm). as promising new animal models, we chose xenopus laevis and three inbred strains of x. tr ...201830390005
size-dependent functional response of xenopus laevis feeding on mosquito larvae.predators can play an important role in regulating prey abundance and diversity, determining food web structure and function, and contributing to important ecosystem services, including the regulation of agricultural pests and disease vectors. thus, the ability to predict predator impact on prey is an important goal in ecology. often, predators of the same species are assumed to be functionally equivalent, despite considerable individual variation in predator traits known to be important for sha ...201830386704
expression of trpv channels during xenopus laevis embryogenesis.transient receptor potential (trp) cation channel genes code for an extensive family of conserved proteins responsible for a variety of physiological processes, including sensory perception, ion homeostasis, and chemical signal transduction. the trp superfamily consists of seven subgroups, one of which is the transient receptor potential vanilloid (trpv) channel family. while trpv channels are relatively well studied in adult vertebrate organisms given their role in functions such as nociception ...201830326274
calcineurin promotes apc/c activation at meiotic exit by acting on both xerp1 and cdc20.vertebrate oocytes await fertilization arrested at metaphase of the second meiotic division. fertilization triggers a transient calcium wave, which induces the activation of the anaphase-promoting complex/cyclosome (apc/c) and its co-activator cdc20 resulting in the destruction of cyclin b and hence meiotic exit. two calcium-dependent enzymes are implicated in fertilization-induced apc/ccdc20 activation: calcium-/calmodulin-dependent kinase type ii (camkii) and calcineurin (can). while the role ...201830373936
assessment of the complex refractive indices of xenopus laevis sciatic nerve for the optimization of optical (nir) neurostimulation.despite an increasing interest in the use of light for neural stimulation, there is little information on how it interacts with neural tissue. the choice of wavelength in most of the optical stimulation literature is based on already available light sources designed for other applications. this paper is the first one to report the complex refractive index of the sciatic nerve of xenopus laevis, which is a crucial parameter for identifying the optimal wavelength of optical stimuli. the xenopus la ...201830371379
cellspecks: a software for automated detection and analysis of calcium channels in live cells.to couple the fidelity of patch-clamp recording with a more high-throughput screening capability, we pioneered a, to our knowledge, novel approach to single-channel recording that we named "optical patch clamp." by using highly sensitive fluorescent ca2+ indicator dyes in conjunction with total internal fluorescence microscopy techniques, we monitor ca2+ flux through individual ca2+-permeable channels. this approach provides information about channel gating analogous to patch-clamp recording at ...201830447989
morphomechanical reactions and mechanically stressed structures in amphibian embryos, as related to gastrulation and axial organs formation.reactions of embryonic tissues to a distributed and concentrated stretching are described and compared with the mechanics of the normal gastrulation movements. a role of mechanically stressed dynamic cell structures in the gastrulation, demarcation of notochord borders and in providing proportionality of the axial rudiments is demonstrated. a morphomechanical scheme of amphibian gastrulation is presented.201830321583
reconstitution of cell-cycle oscillations in microemulsions of cell-free xenopus egg extracts.real-time measurement of oscillations at the single-cell level is important to uncover the mechanisms of biological clocks. although bulk extracts prepared from xenopus laevis eggs have been powerful in dissecting biochemical networks underlying the cell-cycle progression, their ensemble average measurement typically leads to a damped oscillation, despite each individual oscillator being sustained. this is due to the difficulty of perfect synchronization among individual oscillators in noisy bio ...201830320763
synthetic light-activated ion channels for optogenetic activation and inhibition.optogenetic manipulation of cells or living organisms became widely used in neuroscience following the introduction of the light-gated ion channel channelrhodopsin-2 (chr2). chr2 is a non-selective cation channel, ideally suited to depolarize and evoke action potentials in neurons. however, its calcium (ca2+) permeability and single channel conductance are low and for some applications longer-lasting increases in intracellular ca2+ might be desirable. moreover, there is need for an efficient lig ...201830333716
multiple combinations of rdl subunits diversify the repertoire of gaba receptors in the honey bee parasite varroa destructor.in insects, γ-aminobutyric acid (gaba) is the major inhibitory neurotransmitter, and gaba-gated ion channels are the target of different classes of insecticides, including fipronil. we report here the cloning of six subunits (four rdl, one lcch3, and one grd) that constitute the repertoire of the gaba-gated ion channel family of the varroa mite (varroa destructor), a honey bee ectoparasite. we also isolated a truncated grd subunit with a premature stop codon. we found that when expressed in xeno ...201830333227
emerin induces nuclear breakage in xenopus extract and early embryos.emerin is an inner nuclear membrane protein often mutated in emery-dreifuss muscular dystrophy. because emerin has diverse roles in nuclear mechanics, cytoskeletal organization, and gene expression, it has been difficult to elucidate its contribution to nuclear structure and disease pathology. in this study, we investigated emerin's impact on nuclei assembled in xenopus laevis egg extract, a simplified biochemical system that lacks potentially confounding cellular factors and activities. notably ...201830332321
assembly of protein complexes restricts diffusion of wnt3a proteins.members of the wnt protein family play roles in many aspects of embryogenesis and homeostasis. despite their biological significance, characteristics of wnt proteins still remain unclear, mainly due to their insolubility after the removal of serum. here we examine wnt proteins in serum-containing media by using analytical ultracentrifugation with a fluorescence detection system. this analysis reveals that wnt3a assembles into high-molecular-weight complexes that become dissociable by interaction ...201830320232
early redox activities modulate xenopus tail regeneration.redox state sustained by reactive oxygen species (ros) is crucial for regeneration; however, the interplay between oxygen (o2), ros and hypoxia-inducible factors (hif) remains elusive. here we observe, using an optic-based probe (optrode), an elevated and steady o2 influx immediately upon amputation. the spatiotemporal o2 influx profile correlates with the regeneration of xenopus laevis tadpole tails. inhibition of ros production but not ros scavenging decreases o2 influx. inhibition of hif-1α i ...201830327466
the balance of two opposing factors mad and myc regulates cell fate during tissue remodeling.cell proliferation and differentiation are two distinct yet coupled processes in development in diverse organisms. understanding the molecular mechanisms that regulate this process is a central theme in developmental biology. the intestinal epithelium is a highly complex tissue that relies on the coordination of cell proliferation within the crypts and apoptosis mainly at the tip of the villi, preservation of epithelial function through differentiation, and homeostatic cell migration along the c ...201830237868
unusual light-reflecting pigment cells appear in the xenopus neural tube culture system in the presence of guanosine.isolation and culture of xenopus laevis neural tubes resulted in differentiation of melanophores and iridophores from neural crest cells; the differentiated melanophores and iridophores were then maintained in culture for more than 6 months. guanosine has been reported to promote reflecting platelet formation in melanin-producing pigment cells; however, the process of pigment organellogenesis is still unclear. in the present study, unusual light-reflecting pigment cells were observed upon additi ...201830309510
two tier hox collinearity mediates vertebrate axial patterning.a two tier mechanism mediates hox collinearity. besides the familiar collinear chromatin modification within each hox cluster (nanocollinearity), there is also a macrocollinearity tier. individual hox clusters and individual cells are coordinated and synchronized to generate multiscale (macro and nano) collinearity in the early vertebrate embryo. macro-collinearity is mediated by three non-cell autonomous hox-hox interactions. these mediate temporal collinearity in early nom (non-organizer mesod ...201830234110
dscam differentially modulates pre- and postsynaptic structural and functional central connectivity during visual system wiring.proper patterning of dendritic and axonal arbors is a critical step in the formation of functional neuronal circuits. developing circuits rely on an array of molecular cues to shape arbor morphology, but the underlying mechanisms guiding the structural formation and interconnectivity of pre- and postsynaptic arbors in real time remain unclear. here we explore how down syndrome cell adhesion molecule (dscam) differentially shapes the dendritic morphology of central neurons and their presynaptic r ...201830219101
uroplakins play conserved roles in egg fertilization and acquired additional urothelial functions during mammalian divergence.uroplakin (up) tetraspanins and their associated proteins are major mammalian urothelial differentiation products that form unique two-dimensional crystals of 16-nm particles ("urothelial plaques") covering the apical urothelial surface. although uroplakins are highly expressed only in mammalian urothelium and are often referred to as being urothelium specific, they are also expressed in several mouse nonurothelial cell types in stomach, kidney, prostate, epididymis, testis/sperms, and ovary/ooc ...201830303751
retinal tissue preparation for high-resolution live imaging of photoreceptors expressing multiple transgenes.live imaging has become the favorite method in recent years to study the protein transport, localization and dynamics in live cells. protein transport is extremely essential for proper function of photoreceptors. aberration in the proper transport of proteins gives rise to the loss of photoreceptor and blindness. on the other hand, the ease of generation of transgenic xenopus laevis tadpoles and the advantage of high resolution live confocal imaging provide new insight into understanding protein ...201830302320
antibacterial and antibiofilm activity and mode of action of magainin 2 against drug-resistant acinetobacter baumannii.antimicrobial peptides (amps) are promising therapeutic agents for treating antibiotic-resistant bacterial infections. previous studies showed that magainin 2 (isolated from african clawed fogs xenopus laevis) has antimicrobial activity against gram-positive and gram-negative bacteria. the present study was conducted to investigate the antibacterial activity of magainin 2 against acinetobacter baumannii. magainin 2 showed excellent antibacterial activity against a. baumannii strains and high sta ...201830301180
functional analysis of a triplet deletion in the gene encoding the sodium glucose transporter 3, a potential risk factor for adhd.sodium-glucose transporters (sglt) belong to the solute carrier 5 family, which is characterized by sodium dependent transport of sugars and other solutes. in contrast, the human sglt3 (hsglt3) isoform, encoded by slc5a4, acts as a glucose sensor that does not transport sugar but induces membrane depolarization by na+ currents upon ligand binding. whole-exome sequencing (wes) of several extended pedigrees with high density of attention-deficit/hyperactivity disorder (adhd) identified a triplet a ...201830286162
bighead is a wnt antagonist secreted by the xenopus spemann organizer that promotes lrp6 endocytosis.the xenopus laevis embryo has been subjected to almost saturating screens for molecules specifically expressed in dorsal spemann organizer tissue. in this study, we performed high-throughput rna sequencing of ectodermal explants, called animal caps, which normally give rise to epidermis. we analyzed dissociated animal cap cells that, through sustained activation of mapk, differentiate into neural tissue. we also microinjected mrnas for cerberus, chordin, fgf8, bmp4, wnt8, and xnr2, which induce ...201830209221
systematic variation of the benzoylhydrazine moiety of the glun2a selective nmda receptor antagonist tcn-201.glun2a containing n-methyl-d-aspartate receptors (nmdars) are important ion channels in the central nervous system and highly involved in several different neurophysiological but also neuropathophysiological processes. however, current understanding of the contribution of glun2a containing nmdars in these processes is incomplete. therefore, highly selective compounds are required to further investigate these ion channels. in 2010, tcn-201 (2), one of the first selective negative allosteric modul ...201830218911
developmental dependence for functional eye regrowth in xenopus laevis. 201830136686
a review of interventions against fetal alcohol spectrum disorder targeting oxidative stress.fetal alcohol spectrum disorder is caused by maternal ethanol exposure; it causes physical, behavioral, cognitive, and neural impairments (murawski et al., 2015). mechanisms of fasd causing damage are not yet fully elucidated. oxidative stress might be one of its mechanisms (henderson et al., 1995). yet no effective treatment against fasd has been found other than ethanol abstention (long et al., 2010).201830205148
in vitro analyses of novel hcn4 gene mutations.the hyperpolarization-activated cyclic nucleotide-gated cation channel hcn4 contributes significantly to the generation of basic cardiac electrical activity in the sinus node and is a mediator of modulation by β-adrenergic stimulation. heterologous expression of sick sinus syndrome (sss) and bradycardia associated mutations within the human hcn4 gene results in altered channel function. the main aim was to describe the functional characterization of three (two novel and one known) missense mutat ...201830196304
amphibian (xenopus laevis) interleukin-8 (cxcl8): a perspective on the evolutionary divergence of granulocyte chemotaxis.the glutamic acid-leucine-arginine (elr) motif is a hallmark feature shared by mammalian inflammatory cxc chemokines such the granulocyte chemo-attractant cxcl8 (interleukin-8, il-8). by contrast, most teleost fish inflammatory chemokines lack this motif. interestingly, the amphibian xenopus laevis encodes multiple isoforms of cxcl8, one of which (cxcl8a) possesses an elr motif, while another (cxcl8b) does not. these cxcl8 isoforms exhibit distinct expression patterns during frog development and ...201830258441
inward rectifier potassium (kir) channels in the soybean aphid aphis glycines: functional characterization, pharmacology, and toxicology.inward rectifier k+ (kir) channels contribute to a variety of physiological processes in insects and are emerging targets for insecticide development. previous studies on insect kir channels have primarily focused on dipteran species (e.g., mosquitoes, fruit flies). here we identify and functionally characterize kir channel subunits in a hemipteran insect, the soybean aphid aphis glycines, which is an economically important insect pest and vector of soybeans. from the transcriptome and genome of ...201830196125
molecular determinants of α-conotoxin potency for inhibition of human and rat α6β4 nicotinic acetylcholine receptors.nicotinic acetylcholine receptors (nachrs) containing α6 and β4 subunits are expressed by dorsal root ganglion neurons and have been implicated in neuropathic pain. rodent models are often used to evaluate the efficacy of analgesic compounds, but species differences may affect the activity of some nachr ligands. a previous candidate α-conotoxin-based therapeutic yielded promising results in rodent models, but failed in human clinical trials, emphasizing the importance of understanding species di ...201830249616
x-fact: xenopus-fast clearing technique.accessibility and imaging of cell compartments in big specimens are crucial for cellular biological research but also a matter of contention. confocal imaging and tissue clearing on whole organs allow for 3d imaging of cellular structures after being subjected to in-toto immunohistochemistry. lately, the passive clarity technique (pact) has been adapted to clear and immunolabel large specimens or individual organs of several aquatic species. we recently demonstrated tissue clearing on one-week o ...201830151770
dye electroporation and imaging of calcium signaling in xenopus nervous system.electroporation is an efficient method of transferring charged macromolecules into living cells in order to study their morphology, function, and connectivity within neuronal networks. labeling cells with fluorophore-coupled macromolecules can be used to trace projections of whole neuronal ensembles, as well as the fine morphology of single cells. here, we present a protocol to visualize pre- and postsynaptic components of a sensory relay synapse in the brain, using the olfactory system of xenop ...201830151769
quantitative proteomics for xenopus embryos ii, data analysis.the oocytes, embryos, and cell-free lysates of the frog xenopus laevis have emerged as powerful models for quantitative proteomic experiments. in the accompanying paper (chapter 13) we describe how to prepare samples and acquire multiplexed proteomics spectra from those. as an illustrative example we use a 10-stage developmental time series from the egg to stage 35 (just before hatching). here, we outline how to convert the ~700,000 acquired mass spectra from this time series into protein expres ...201830151768
evaluating iodide recycling inhibition as a novel molecular initiating event for thyroid axis disruption in amphibians.the enzyme iodotyrosine deiodinase (dehalogenase, iyd) catalyzes iodide recycling and promotes iodide retention in thyroid follicular cells. loss of function or chemical inhibition of iyd reduces available iodide for thyroid hormone synthesis, which leads to hormone insufficiency in tissues and subsequent negative developmental consequences. iyd activity is especially critical under conditions of lower dietary iodine and in low iodine environments. our objective was to evaluate the toxicological ...201830137636
kv12.1 channels are not sensitive to gqpcr-triggered activation of phospholipase cβ.kv12.1 k+ channels are expressed in several brain areas, but no physiological function could be attributed to these subunits so far. as genetically-modified animal models are not available, identification of native kv12.1 currents must rely on characterization of distinct channel properties. recently, it was shown in xenopus laevis oocytes that kv12.1 channels were modulated by membrane pi(4,5)p2. however, it is not known whether these channels are also sensitive to physiologically-relevant pi(4 ...201830136882
cross-limb communication during xenopus hindlimb regenerative response: non-local bioelectric injury signals.regeneration of damaged body parts requires coordination of size, shape, location and orientation of tissue with the rest of the body. it is not currently known to what extent injury sites communicate with the remaining soma during repair, or what information may emanate from the injury site and reach other regions. we examined the bioelectric properties (resting potential gradients in the epidermis) of xenopus laevis froglets undergoing hindlimb amputation and observed that the contralateral (u ...201830126906
draft genome of dugesia japonica provides insights into conserved regulatory elements of the brain restriction gene nou-darake in planarians.planarians are non-parasitic platyhelminthes (flatworms) famous for their regeneration ability and for having a well-organized brain. dugesia japonica is a typical planarian species that is widely distributed in the east asia. extensive cellular and molecular experimental methods have been developed to identify the functions of thousands of genes in this species, making this planarian a good experimental model for regeneration biology and neurobiology. however, no genome-level information is ava ...201830181897
the cytoskeletal protein zyxin interacts with the zinc-finger transcription factor zic1 and plays the role of a scaffold for gli1 and zic1 interactions during early development of xenopus laevis.we have shown recently that the cytoskeletal protein zyxin participates in the fine tuning of the neural plate pattering in xenopus laevis embryos by modulating activity of one of the effectors of hedgehog (shh) signaling cascade, the transcription factor gli1. in the present work, we show that zyxin can also interact with the potential modulator of the shh pathway, the transcription factor zic1. the interaction of proteins occurs primarily by mean of the zinc-finger domain of zic1 and 2nd lim d ...201830180953
a single xlf dimer bridges dna ends during nonhomologous end joining.nonhomologous end joining (nhej) is the primary pathway of dna double-strand-break repair in vertebrate cells, yet how nhej factors assemble a synaptic complex that bridges dna ends remains unclear. to address the role of xrcc4-like factor (xlf) in synaptic-complex assembly, we used single-molecule fluorescence imaging in xenopus laevis egg extract, a system that efficiently joins dna ends. we found that a single xlf dimer binds dna substrates just before the formation of a ligation-competent sy ...201830177755
ras-dva small gtpases lost during evolution of amniotes regulate regeneration in anamniotes.in contrast to amniotes (reptiles, birds and mammals), anamniotes (fishes and amphibians) can effectively regenerate body appendages such as fins, limbs and tails. why such a useful capability was progressively lost in amniotes remains unknown. as we have hypothesized recently, one of the reasons for this could be loss of some genes regulating the regeneration in evolution of amniotes. here, we demonstrate the validity of this hypothesis by showing that genes of small gtpases ras-dva1 and ras-dv ...201830158598
isolation and primary culture methods of adult and larval myogenic cells from xenopus laevis.during amphibian metamorphosis, larval-to-adult conversion of the myogenic system occurs and there are two distinct types of muscle stem cells; larval myogenic cells have a death-fate by apoptosis in the presence of thyroid hormone t3, and adult myogenic cells have a life-fate under the same conditions. here, we describe isolation and culture methods for adult and larval myogenic cells from the frog, xenopus laevis. both types of cultured myogenic cells undergo cell division and cell differentia ...201830151775
isolation and culture of amphibian (xenopus laevis) sub-capsular liver and bone marrow cells.the x. laevis sub-capsular liver is thought to be the principal hematopoietic site of xenopodinae species from early development and, in case of certain species, into adulthood. the xenopus bone marrow appears to be comprised of precursor cells committed to myeloid lineages, such as macrophage- and granulocyte-progenitor cells. with alarming increases in the contribution of pathogenic infections to the global amphibian declines, now more than ever a better understanding of the mechanisms control ...201830151774
quantitative proteomics of xenopus embryos i, sample preparation.xenopus oocytes and embryos are model systems optimally suited for quantitative proteomics. this is due to the availability of large amount of protein material and the ease of physical manipulation. furthermore, facile in vitro fertilization provides superbly synchronized embryos for cell cycle and developmental stages. here, we detail protocols developed over the last few years for sample preparation of multiplexed proteomics with tmt-tags followed by quantitative mass spectrometry analysis usi ...201830151767
a simple knock-in system for xenopus via microhomology mediated end joining repair.following completion of the genome sequences of xenopus tropicalis and x. laevis, gene targeting techniques have become increasingly important for the further development of xenopus research in the life sciences. gene knockout using programmable nucleases, such as talen and crispr/cas9, has reached a level whereby we can readily and routinely perform loss-of-function analysis of genes of interest in these species. however, there is still room for improvement in gene knock-in techniques owing to ...201830151761
generation and care of xenopus laevis and xenopus tropicalis embryos.robust and efficient protocols for fertilization and early embryo care of xenopus laevis and xenopus tropicalis are essential for experimental success, as well as maintenance and propagation of precious animal stocks. the rapid growth of the national xenopus resource has required effective implementation and optimization of these protocols. here, we discuss the procedures used at the national xenopus resource, which we found helpful for generation and early upkeep of xenopus embryos and tadpoles ...201830151756
husbandry, general care, and transportation of xenopus laevis and xenopus tropicalis.maintenance of optimal conditions such as water parameters, diet, and feeding is essential to a healthy xenopus laevis and xenopus tropicalis colony and thus to the productivity of the lab. our prior husbandry experience as well as the rapid growth of the national xenopus resource has given us a unique insight into identifying and implementing these optimal parameters into our husbandry operations. here, we discuss our standard operating procedures that will be of use to both new and established ...201830151755
photocatalytic degradation of azo dye using core@shell nano-tio2 particles to reduce toxicity.clean and safe water is fundamental for human and environmental health. traditional remediation of textile dye-polluted water with chemical, physical, and biological processes has many disadvantages. due to this, nano-engineered materials are drawing more attention to this area. however, the widespread use of nano-particles for this purpose may lead to photocatalytic degradation of xenobiotics, while increasing the risk of nano-particle-induced ecotoxicity. therefore, we comparatively evaluated ...201830136182
a dual-app nucleoside probe provides structural insights into the human telomeric overhang in live cells.understanding the topology adopted by individual g-quadruplex (gq)-forming sequences in vivo and targeting a specific gq motif among others in the genome will have a profound impact on gq-directed therapeutic strategies. however, this remains a major challenge as most of the tools poorly distinguish different gq conformations and are not suitable for both cell-free and in-cell analysis. here, we describe an innovative probe design to investigate gq conformations and recognition in both cell-free ...201830192541
multiscale analysis of architecture, cell size and the cell cortex reveals cortical f-actin density and composition are major contributors to mechanical properties during convergent extension.the large-scale movements that construct complex three-dimensional tissues during development are governed by universal physical principles. fine-grained control of both mechanical properties and force production is crucial to the successful placement of tissues and shaping of organs. embryos of the frog xenopus laevis provide a dramatic example of these physical processes, as dorsal tissues increase in young's modulus by six-fold to 80 pascal over 8 h as germ layers and the central nervous syst ...201830190279
tectal crfr1 receptors modulate food intake and feeding behavior in the south african clawed frog xenopus laevis.the optic tectum and superior colliculus rapidly inhibit food intake when a visual threat is present. previous work indicates that crf, acting on crfr1 receptors, may play a role in tectal inhibition of feeding behavior and food intake. here we test the hypothesis that tectal crfr1 receptors modulate food intake and feeding behavior in juvenile xenopus laevis. we performed five experiments to test the following questions: 1) does tectal crf injection decrease food intake/feeding behavior? 2) doe ...201830077740
molecular characterization of wdr68 gene in embryonic development of xenopus laevis.wdr68, also known as dcaf7, is a wd40 repeated domain protein highly conserved in eukaryotic organisms in both plants and animals. this protein participates in numerous cellular processes and exerts its function through interaction with other proteins. in the present work, we isolated, sequenced and characterized cdna corresponding to the wdr68 gene in embryos of the amphibian xenopus laevis. syntenic analysis revealed high conservation of the genomic region containing the wdr68 locus in amniote ...201830125741
in vitro expression and functional characterization of npa motifs in aquaporins of nosema bombycis.nosema bombycis contains functional aquaporins (nbaqps), which are key targets for exploring the mechanism of n. bombycis infection; however, the regulation of these nbaqps remains unknown. the two highly conserved asparagine-proline-alanine sequences (npa motifs) play important roles in aqp biogenesis. as part of this study, we constructed a series of nbaqp mutants (nbaqp_npa1, nbaqp_npa2, and nbaqp_npa1,2) and expressed them in bmn cells. the results showed that mutations in either npa motif o ...201830105406
polystyrene microplastics did not affect body growth and swimming activity in xenopus laevis tadpoles.a growing number of studies have highlighted the contamination and the effects towards organisms of diverse microplastics (μps) in the marine environment. surprisingly, although the main sources of μps for marine environments are inland surface waters, the information on the occurrence and the effects of μps in freshwater ecosystems is still scant. thus, the aim of the present work is to investigate the ingestion and possible adverse effects due to the exposure to polystyrene μps (psμps; ø = 3 μ ...201830317408
trypanosoma brucei aquaglyceroporins mediate the transport of metabolic end-products: methylglyoxal, d-lactate, l-lactate and acetate.bloodstream forms of trypanosoma (t.) brucei, the causative agent of african sleeping sickness, possess a highly active glycolysis, which generates as main end-products: pyruvate under aerobic conditions, and pyruvate and glycerol under anaerobic conditions. to secrete them into the extracellular milieu, the parasites have at least two main specific membrane proteins, the pyruvate transporter and the aquaglyceroporins however, there are several other minor products from the glycolysis that must ...201830409521
brief local application of progesterone via a wearable bioreactor induces long-term regenerative response in adult xenopus hindlimb.the induction of limb repair in adult vertebrates is a pressing, unsolved problem. here, we characterize the effects of an integrated device that delivers drugs to severed hindlimbs of adult xenopus laevis, which normally regenerate cartilaginous spikes after amputation. a wearable bioreactor containing a silk protein-based hydrogel that delivered progesterone to the wound site immediately after hindlimb amputation for only 24 hr induced the regeneration of paddle-like structures in adult frogs. ...201830404012
differential modulation of human gabac-ρ1 receptor by sulfur-containing compounds structurally related to taurine.the amino acid taurine (2-aminoethanesulfonic acid) modulates inhibitory neurotransmitter receptors. this study aimed to determine if the dual action of taurine on gabac-ρ1r relates to its structure. to address this, we tested the ability of the structurally related compounds homotaurine, hypotaurine, and isethionic acid to modulate gabac-ρ1r.201830075755
apoptosis propagates through the cytoplasm as trigger waves.apoptosis is an evolutionarily conserved form of programmed cell death critical for development and tissue homeostasis in animals. the apoptotic control network includes several positive feedback loops that may allow apoptosis to spread through the cytoplasm in self-regenerating trigger waves. we tested this possibility in cell-free xenopus laevis egg extracts and observed apoptotic trigger waves with speeds of ~30 micrometers per minute. fractionation and inhibitor studies implicated multiple f ...201830093599
the age-regulated zinc finger factor znf367 is a new modulator of neuroblast proliferation during embryonic neurogenesis.global population aging is one of the major social and economic challenges of contemporary society. during aging the progressive decline in physiological functions has serious consequences for all organs including brain. the age-related incidence of neurodegenerative diseases coincides with the sharp decline of the amount and functionality of adult neural stem cells. recently, we identified a short list of brain age-regulated genes by means of next-generation sequencing. among them znf367 codes ...201830087422
gli2 is required for the induction and migration of xenopus laevis neural crest.the neural crest (nc) is a multipotent migratory embryonic population that is formed during late gastrulation and gives rise to a wide array of derivatives, including cells from the peripheral nervous system (pns), the craniofacial bones and cartilages, peripheral glial cells, and melanocyte cells, among others. in this work we analyzed the role of the hedgehog signaling pathway effector gli2 in xenopus nc. we provide evidence that the gli2 gene is expressed in the prospective, premigratory and ...201830086335
spatial and temporal analysis of pcp protein dynamics during neural tube closure.planar cell polarity (pcp) controls convergent extension and axis elongation in all vertebrates. although asymmetric localization of pcp proteins is central to their function, we understand little about pcp protein localization during convergent extension. here, we use quantitative live imaging to simultaneously monitor cell intercalation behaviors and pcp protein dynamics in the xenopus laevis neural plate epithelium. we observed asymmetric enrichment of pcp proteins, but more interestingly, we ...201830080139
rare coding variants in genes encoding gabaa receptors in genetic generalised epilepsies: an exome-based case-control study.genetic generalised epilepsy is the most common type of inherited epilepsy. despite a high concordance rate of 80% in monozygotic twins, the genetic background is still poorly understood. we aimed to investigate the burden of rare genetic variants in genetic generalised epilepsy.201830033060
a simple decision to move in response to touch reveals basic sensory memory and mechanisms for variable response times.short-term working memory and decision-making are usually studied in the cerebral cortex; in many models of simple decision making, sensory signals build slowly and noisily to threshold to initiate a motor response after long, variable delays. when touched, hatchling frog tadpoles decide whether to swim; we define the long and variable delays to swimming and use whole-cell recordings to uncover the neurons and processes responsible. firing in sensory and sensory pathway neurons is short latency, ...201830074236
disruption of amino acid homeostasis by novel asct2 inhibitors involves multiple targets.the glutamine transporter asct2 (slc1a5) is actively investigated as an oncological target, but the field lacks efficient asct2 inhibitors. a new group of asct2 inhibitors, 2-amino-4-bis(aryloxybenzyl)aminobutanoic acids (aaba), were developed recently and shown to suppress tumor growth in preclinical in vivo models. to test its specificity, we deleted asct2 in two human cancer cell lines. surprisingly, growth of parental and asct2-knockout cells was equally sensitive to aaba compounds. aaba com ...201830072900
tight junction-associated protein gef-h1 in the neighbours of dividing epithelial cells is essential for adaptation of cell-cell membrane during cytokinesis.animal cells divide by a process called cytokinesis which relies on the constriction of a contractile actomyosin ring leading to the production of two daughter cells. cytokinesis is an intrinsic property of cells which occurs even for artificially isolated cells. during division, isolated cells undergo dramatic changes in shape such as rounding and membrane deformation as the division furrow ingresses. however, cells are often embedded in tissues and thus are surrounded by neighbouring cells. ho ...201830056063
nosip functions during vertebrate eye and cranial cartilage development.the nitric oxide synthase interacting protein (nosip) has been associated with diverse human diseases including psychological disorders. in line, early neurogenesis of mouse and xenopus is impaired upon nosip deficiency. nosip knockout mice show craniofacial defects and the down-regulation of nosip in the mouse and xenopus leads to microcephaly. until now, the exact underlying molecular mechanisms of these malformations were still unknown.201830055071
a cyclic nucleotide-gated channel, hvcngc2-3, is activated by the co-presence of na⁺ and k⁺ and permeable to na⁺ and k⁺ non-selectively.cyclic nucleotide-gated channels (cngcs) have been postulated to contribute significantly in plant development and stress resistance. however, their electrophysiological properties remain poorly understood. here, we characterized barley cngc2-3 (hvcngc2-3) by the two-electrode voltage-clamp technique in the xenopus laevis oocyte heterologous expression system. current was not observed in x. laevis oocytes injected with hvcngc2-3 complementary rna (crna) in a bathing solution containing either na ...201830049942
multiple massive domestication and recent amplification of kolobok superfamily transposons in the clawed frog xenopus.dna transposons are generally destroyed by mutations and have short lifespans in hosts, as they are neutral or harmful to the host and therefore not conserved by natural selection. the clawed frog xenopus harbors many dna transposons and certain families, such as t2-mite, have extremely long lives. these have ancient origins, but have shown recent transposition activity. in addition, certain transposase genes may have been "domesticated" by xenopus and conserved over long time periods by natural ...201829946483
adamts9, a member of the adamts family, in xenopus development.extracellular matrix (ecm) remodeling by metalloproteinases is crucial during development. the adamts (a disintegrin and metalloproteinase with thrombospondin type i motifs) enzymes are secreted, multi-domain matrix-associated zinc metalloendopeptidases that have diverse roles in tissue morphogenesis and patho-physiological remodeling. the human family includes 19 members. in this study we identified the 19 members of the adamts family in xenopus laevis and xenopus tropicalis. gene identificatio ...201829935379
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