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determination of metals and pharmaceutical compounds released in hospital wastewater from toluca, mexico, and evaluation of their toxic impact.due to the activities inherent to medical care units, the hospital effluent released contains diverse contaminants such as tensoactives, disinfectants, metals, pharmaceutical products and chemical reagents, which are potentially toxic to the environment since they receive no treatment or are not effectively removed by such treatment before entering the drain. they are incorporated into municipal wastewater, eventually entering water bodies where they can have harmful effects on organisms and can ...201829751329
yokukansan contains compounds that antagonize the 5-ht3 receptor.we recently focused on alkaloids in uncaria hook (a constituent of the kampo medicine, yokukansan) and identified the pharmacological action of geissoschizine methyl ether on several g protein-coupled receptors. however, the functions of other identified alkaloids in uncaria hook, including hirsutine, hirsuteine, rhynchophylline, isorhynchophylline, corynoxeine, isocorynoxeine, are not clear.201829747744
foxo4 activity is regulated by phosphorylation and the cellular environment during dehydration in the african clawed frog, xenopus laevis.the african clawed frog, xenopus laevis, is capable of enduring seasonal bouts of severe dehydration stress resulting from transcriptional regulation that facilitates a pro-survival response. previous studies have shown that dehydration increases antioxidant gene expression in this amphibian. as foxo4 is known to regulate several antioxidant genes, we sought to understand how differential phosphorylation and environmental factors (urea, temperature) may contribute to its transcriptional regulati ...201829746959
deciphering the molecular determinants of cholinergic anthelmintic sensitivity in nematodes: when novel functional validation approaches highlight major differences between the model caenorhabditis elegans and parasitic species.cholinergic agonists such as levamisole and pyrantel are widely used as anthelmintics to treat parasitic nematode infestations. these drugs elicit spastic paralysis by activating acetylcholine receptors (achrs) expressed in nematode body wall muscles. in the model nematode caenorhabditis elegans, genetic screens led to the identification of five genes encoding levamisole-sensitive-achr (l-achr) subunits: unc-38, unc-63, unc-29, lev-1 and lev-8. these subunits form a functional l-achr when hetero ...201829719008
transposable elements and polyploid evolution in animals.polyploidy in animals is much less common than in plants, where it is thought to be pervasive in all higher plant lineages. recent studies have highlighted the impact of polyploidization and the associated process of diploidy restoration on the evolution and speciation of selected taxonomic groups in the animal kingdom: from vertebrates represented by salmonid fishes and african clawed frogs to invertebrates represented by parasitic root-knot nematodes and bdelloid rotifers. in this review, we f ...201829715568
cardiac regeneration in xenopus tropicalis and xenopus laevis: discrepancies and problems.two studies have recently focused on adult heart regeneration in xenopus. while we reported on cardiac myogenic regeneration in xenopus tropicalis after injury, marshall and colleagues found no regeneration in an injured heart in xenopus laevis. here, we would like to join the discussion initiated by marshall et al. who debated the methods and species differences in both studies. we agree with their view that the species difference in cardiac regenerative capacity could lead to different results ...201829713455
is adult cardiac regeneration absent in xenopus laevis yet present in xenopus tropicalis?we recently used an endoscopy-based resection method to explore the consequences of cardiac injury in adult xenopus laevis, obtaining the result that the adult xenopus heart is unable to regenerate. at 11 months post-amputation, cellular and biological marks of scarring persisted. we thus concluded that, contrary to urodeles and teleosts, adult anurans share a cardiac injury outcome similar to adult mammals. however, in their work published in this journal on the 13 december 2017, liao et al. sh ...201829713454
nmda receptor signaling is important for neural tube formation and for preventing antiepileptic drug-induced neural tube defects.failure of neural tube closure leads to neural tube defects (ntds), which can have serious neurological consequences or be lethal. use of antiepileptic drugs (aeds) during pregnancy increases the incidence of ntds in offspring by unknown mechanisms. here we show that during xenopus laevis neural tube formation, neural plate cells exhibit spontaneous calcium dynamics that are partially mediated by glutamate signaling. we demonstrate that nmda receptors are important for the formation of the neura ...201829712790
xenopus laevis oocyte as a model for the study of the cytoskeleton.at the beginning of diplotene, the oocyte of xenopus laevis is a cell of about 10-20 microns destined to increase 10,000-fold its size when the oocyte becomes filled with yolk platelets and has accumulated a great number of pigment granules in a half of its periphery. its internal architecture is gradually accomplished during growth because of several factors, especially because of cytoskeletal changes. in the fully-grown oocyte, the cytoskeleton appears to sustain the eccentrically located germ ...201829705198
roles of xenopus chemokine ligand cxclh (xcxclh) in early embryogenesis.several chemokine molecules control cell movements during early morphogenesis. however, it is unclear whether chemokine molecules affect cell fate. here, we identified and characterized the cxc-type chemokine ligand in xenopus laevis, xenopus cxclh (xcxclh), during early embryogenesis. xcxclh is expressed in the dorsal vegetal region at the gastrula stage. both overexpression and knockdown of xcxclh in the dorsal region inhibited gastrulation. xcxclh contributed to the attraction of mesendoderma ...201829700804
effect of hemin, baicalein and heme oxygenase-1 (ho-1) enzyme activity inhibitors on cd-induced accumulation of ho-1, hsps and aggresome-like structures in xenopus kidney epithelial cells.cadmium is a highly toxic environmental pollutant that can cause many adverse effects including cancer, neurological disease and kidney damage. aquatic amphibians are particularly susceptible to this toxicant as it was shown to cause developmental abnormalities and genotoxic effects. in mammalian cells, the accumulation of heme oxygenase-1 (ho-1), which catalyzes the breakdown of heme into co, free iron and biliverdin, was reported to protect cells against potentially lethal concentrations of cd ...201829698685
microvascular anatomy of the brain of the adult pipid frog, xenopus laevis (daudin): a scanning electron microscopic study of vascular corrosion casts.to demonstrate the 3d microvascular anatomy of the brain of the model organism xenopus laevis daudin scanning electron microscopy of vascular corrosion casts was correlated with light microscopy of stained 7 µm thick serial tissues sections. results showed that supplying arteries descended from the leptomeningeal surface without remarkable branchings straight to the subventricular zone where they branched and capillarized. capillaries showed few h- and/or y-shaped anastomoses during their centri ...201829693258
developmental toxicity of dibutyl phthalate and citrate ester plasticizers in xenopus laevis embryos.citrate esters have been considered as alternatives to phthalate plasticizers. being considered to have low toxicity in mammals, their toxicological information for aquatic animals remains poorly understood. we examined the developmental toxicity of citrate esters including tributyl o-acetylcitrate (atbc), triethyl 2-acetylcitrate (atec), and trihexyl o-acetylcitrate (athc) together with dibutyl phthalate (dbp) based on the frog embryo teratogenesis assay-xenopus (fetax). atbc has the lowest 96  ...201829684872
new apetx-like peptides from sea anemone heteractis crispa modulate asic1a channels.sea anemones are an abundant source of various biologically active peptides. the hydrophobic 20% ethanol fraction of tropical sea anemone heteractis crispa was shown to contain at least 159 peptide compounds including neurotoxins, proteinase and α-amylase inhibitors, as well as modulators of acid-sensing ion channels (asics). the three new peptides, π-anmtx hcr 1b-2, -3, and -4 (41 aa) (short names hcr 1b-2, -3, -4), identified by a combination of reversed-phase liquid chromatography and mass sp ...201829684594
comparison of oocyte mrna localization patterns in sterlet acipenser ruthenus and african clawed frog xenopus laevis.in oocytes, rna localization has critical implications, as assembly of proteins in particular subcellular domains is crucial to embryo development. the distribution of mrna molecules can identify and characterize localized transcripts. the goal of this study was to clarify the origin of primordial germ cells in the oocyte body plan and to reveal the generation of cell lineages by localized rnas. the distribution of 12 selected mrnas in sterlet acipenser ruthenus oocytes was investigated by qpcr ...201829682883
effects of multiple life stage exposure to the fungicide prochloraz in xenopus laevis: manifestations of antiandrogenic and other modes of toxicity.the larval amphibian growth and development assay (lagda) is an internationally harmonized testing guideline for evaluating effects of chronic chemical exposure in amphibians. in order to evaluate the effects of chronic exposure to an antiandrogenic chemical in an amphibian model, prochloraz was tested using a variation of the lagda design. exposure was initiated with <1d post-fertilization embryos at nominal concentrations of 0, 6.7, 20, 60 and 180 μg/l (0, 18, 53, 159, 478 nm) and continued in ...201829674245
fluorescent anesthetics.methods for using exogenous fluorophore and general anesthetic 1-aminoanthracene (1-ama) and its photoactive derivative 1-azidoanthracene (1-aza) are provided. 1-ama potentiates gabaa chloride currents and immobilizes xenopus laevis tadpoles. cellular and tissue anesthetic distribution can be imaged for quantifying "on-pathway" and "off-pathway" targets. 1-aza shares targets with 1-ama and offers further optoanesthetic spatial and temporal control upon near-uv laser irradiation. furthermore, 1-a ...201829673536
galantamine is not a positive allosteric modulator of human α4β2 or α7 nicotinic acetylcholine receptors.the alkaloid galantamine was originally isolated from the green snowdrop galanthus woronowii and is currently marketed as a drug for treatment of mild to moderate dementia in patients with alzheimer's disease. in addition to a well-documented proficiency to inhibit acetylcholinesterase, galantamine has been reported to increase neuronal nicotinic ach (nach) receptor function by acting as a positive allosteric modulator. yet there remains controversy regarding these findings in the literature. to ...201829669164
atopt6 protein functions in long-distance transport of glutathione in arabidopsis thaliana.the involvement of the arabidopsis oligopeptide transporter atopt6, which was previously shown to take up glutathione (gsh) when expressed in yeast cells or in xenopus laevis oocytes, in gsh transport was analyzed using opt6 knockout mutant lines. the concentration of gsh in flowers or siliques was lower in opt6 mutants relative to wild-type plants, suggesting involvement of atopt6 in long-distance transport of gsh. the gsh concentration in phloem sap was similar between opt6 mutants and wild-ty ...201829669129
developmental expression profiles and thyroidal regulation of cytokines during metamorphosis in the amphibian xenopus laevis.early life-stages of amphibians rely on the innate immune system for defense against pathogens. while thyroid hormones (th) are critical for metamorphosis and later development of the adaptive immune system, the role of th in innate immune system development is less clear. an integral part of the innate immune response are pro-inflammatory cytokines - effector molecules that allow communication between components of the immune system. the objective of this study was to characterize the expressio ...201829656046
new targets for old drugs: cardiac glycosides inhibit atrial-specific k2p3.1 (task-1) channels.cardiac glycosides have been used in the treatment of arrhythmias for more than 200 years. two-pore-domain (k2p) potassium channels regulate cardiac action potential repolarization. recently, k2p3.1 [tandem of p domains in a weak inward rectifying k+ channel (twik)-related acid-sensitive k+ channel (task)-1] has been implicated in atrial fibrillation pathophysiology and was suggested as an atrial-selective antiarrhythmic drug target. we hypothesized that blockade of cardiac k2p channels contribu ...201829643254
expression of the inactivating deiodinase, deiodinase 3, in the pre-metamorphic tadpole retina.thyroid hormone (th) orchestrates amphibian metamorphosis. thus, this developmental phase is often used to study th-dependent responses in specific tissues. however, th signaling appears early in development raising the question of the control of th availability in specific cell types prior to metamorphosis. th availability is under strict temporal and tissue-specific control by deiodinases. we examined the expression of the th-inactivating enzyme, deiodinase type 3 (d3), during early retinal de ...201829641587
tbx2 is required for the suppression of mesendoderm during early xenopus development.t-box family proteins are dna-binding transcriptional regulators that play crucial roles during germ layer formation in the early vertebrate embryo. well-characterized members of this family, including the transcriptional activators brachyury and vegt, are essential for the proper formation of mesoderm and endoderm, respectively. to date, t-box proteins have not been shown to play a role in the promotion of the third primary germ layer, ectoderm.201829633424
an ex vivo assay for screening glucocorticoid signaling disruption based on glucocorticoid-response gene transcription in xenopus tails.there is a pressing need for developing in vivo or ex vivo assays to screen the glucocorticoid (gc) signaling disruption of chemicals. thus, we aimed to establish an ex vivo assay for screening gc signaling disruption based on the gc-response gene transcription in xenopus laevis tails cultured ex vivo. firstly, we investigated effects of corticosterone (cort, a main gc in frogs) on gc-response gene expression, and determined the six genes as molecular endpoints for assaying the gc signaling disr ...201829628076
micrornp-mediated translational activation of nonadenylated mrnas in a mammalian cell-free system.micrornas are small noncoding rnas that regulate translation and mrna stability by binding target mrnas in complex with argonaute (ago) proteins. ago interacts with a member of the tnrc6 family proteins to form a micrornp complex, which recruits the ccr4-not complex to accelerate deadenylation and inhibits translation. micrornas primarily repress translation of target mrnas but have been shown to enhance translation of a specific type of target reporter mrnas in various experimental systems: g0 ...201829626383
an intersubunit electrostatic interaction in the gabaa receptor facilitates its responses to benzodiazepines.benzodiazepines are positive allosteric modulators of the gabaa receptor (gabaar), acting at the α-γ subunit interface to enhance gabaar function. gaba or benzodiazepine binding induces distinct conformational changes in the gabaar. the molecular rearrangements in the gabaar following benzodiazepine binding remain to be fully elucidated. using two molecular models of the gabaar, we identified electrostatic interactions between specific amino acids at the α-γ subunit interface that were broken by ...201829622679
aluminum-activated malate transporters can facilitate gaba transport.plant aluminum-activated malate transporters (almts) are currently classified as anion channels; they are also known to be regulated by diverse signals, leading to a range of physiological responses. gamma-aminobutyric acid (gaba) regulation of anion flux through almt proteins requires a specific amino acid motif in almts that shares similarity with a gaba binding site in mammalian gabaa receptors. here, we explore why taalmt1 activation leads to a negative correlation between malate efflux and ...201829618628
new information on morphology and molecular data of camallanid nematodes parasitising xenopus laevis (anura: pipidae) in south africa.three species of nematodes from the camallanidae that are known to infect xenopus laevis daudin (anura: pipidae) were collected from several localities across south africa. new data on morphology, partial 28s and cox1 genes, infection levels and distribution are presented herein. the most common species, batrachocamallanus slomei southwell et kirshner, 1937, from the stomach and less often oesophagus, was found in eight localities. camallanus kaapstaadi southwell et kirshner, 1937, also from the ...201829611533
distinct mhc class i-like interacting invariant t cell lineage at the forefront of mycobacterial immunity uncovered in xenopus.the amphibian xenopus laevis is to date the only species outside of mammals where a mhc class i-like (mhc-like) restricted innate-like (i) t cell subset (ivα6 t cells) reminiscent of cd1d-restricted inkt cells has been identified and functionally characterized. this provides an attractive in vivo model to study the biological analogies and differences between mammalian it cells and the evolutionarily antecedent xenopus it cell defense system. here, we report the identification of a unique it cel ...201829610296
divergent evolutionary trajectories of two young, homomorphic, and closely related sex chromosome systems.there exists extraordinary variation among species in the degree and nature of sex chromosome divergence. however, much of our knowledge about sex chromosomes is based on comparisons between deeply diverged species with different ancestral sex chromosomes, making it difficult to establish how fast and why sex chromosomes acquire variable levels of divergence. to address this problem, we studied sex chromosome evolution in two species of african clawed frog (xenopus), both of whom acquired novel ...201829608717
preparations and protocols for whole cell patch clamp recording of xenopus laevis tectal neurons.the xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (rgcs) in the eye which form synapses directly onto neurons in the optic tectum, is a popular model to study how neural circuits self-assemble. the ability to carry out whole cell patch clamp recordings from tectal neurons and to record rgc-evoked responses, either in vivo or using a whole brain preparation, has generated a large body of high-resolution data about the mechanisms underlying normal, and abnormal, cir ...201829608176
xenopus laevis macrophage-like cells produce xcl-1, an intelectin family serum lectin that recognizes bacteria.xenopus laevis ca2+ -dependent lectin-1 (xcl-1) is an intelectin family serum lectin that selectively recognizes carbohydrate chains on the bacterial cell surface. immunofluorescence examination of control spleen tissues from normal x. laevis revealed cells producing xcl-1 (xcl-1+ cells) exclusively in red pulps. intraperitoneal injection of escherichia coli lipopolysaccharide (lps) caused a marked increase in the number of xcl-1+ cells in red pulps on day 3, followed by a rapid decrease to near ...201829604105
cellular composition and organization of the spinal cord central canal during metamorphosis of the frog xenopus laevis.studying the cellular composition and morphological changes of cells lining the central canal during xenopus laevis metamorphosis could contribute to understand postnatal development and spinal cord regeneration. here we report the analysis of central canal cells at different stages during metamorphosis using immunofluorescence for protein markers expression, transmission and scanning electron microscopy and cell proliferation assays. the central canal was regionalized according to expression of ...201829603210
effects of environmentally relevant sub-chronic atrazine concentrations on african clawed frog (xenopus laevis) survival, growth and male gonad development.sub-chronic toxicity of environmentally relevant atrazine concentrations on exposed tadpoles and adult male african clawed frogs (xenopus laevis) was evaluated in a quality controlled laboratory for 90 days. the aim of this study was to determine the effects of atrazine on the survival, growth and gonad development of african clawed frogs. after exposure of tadpoles to atrazine concentrations of 0 (control), 0.01, 200 and 500 μg l-1 in water, mortality rates of 0, 0, 3.3 and 70% respectively wer ...201829602044
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
mir-206 is required for changes in cell adhesion that drive muscle cell morphogenesis in xenopus laevis.micrornas (mirnas) are highly conserved small non-coding rna molecules that post-transcriptionally regulate gene expression in multicellular organisms. within the set of muscle-specific mirnas, mir-206 expression is largely restricted to skeletal muscle and is found exclusively within the bony fish lineage. although many studies have implicated mir-206 in muscle maintenance and disease, its role in skeletal muscle development remains largely unknown. here, we examine the role of mir-206 during x ...201829596841
the degenerin region of the human bile acid-sensitive ion channel (basic) is involved in channel inhibition by calcium and activation by bile acids.the bile acid-sensitive ion channel (basic) is a member of the enac/degenerin family of ion channels. it is activated by bile acids and inhibited by extracellular ca2+. the aim of this study was to explore the molecular mechanisms mediating these effects. the modulation of basic function by extracellular ca2+ and tauro-deoxycholic acid (t-dca) was studied in xenopus laevis oocytes heterologously expressing human basic using the two-electrode voltage-clamp and outside-out patch-clamp techniques. ...201829589117
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
des-formylflustrabromine (dfbr): a structure-activity study on its ability to potentiate the action of acetylcholine at α4β2 nicotinic acetylcholine receptors.the naturally occurring indole alkaloid des-formylflustrabromine (dfbr; 1) is one of the first agents shown to act as a selective positive allosteric modulator (pam) at α4β2 nicotinic acetylcholine receptors (nachrs). we previously deconstructed this agent to determine which of its structural features contribute to its actions and have identified an agent that might serve as the basis for a " working pharmacophore". here, we elaborate the dfbr (1; ec50 = 0.2 μm) structure to identify how various ...201830028943
transplantation of ears provides insights into inner ear afferent pathfinding properties.numerous tissue transplantations have demonstrated that otocysts can develop into normal ears in any location in all vertebrates tested thus far, though the pattern of innervation of these transplanted ears has largely been understudied. here, expanding on previous findings that transplanted ears demonstrate capability of local brainstem innervation and can also be innervated themselves by efferents, we show that inner ear afferents grow toward the spinal cord mostly along existing afferent and ...201830027559
amphibian (xenopus laevis) tadpoles and adult frogs differ in their use of expanded repertoires of type i and type iii interferon cytokines.while amphibians around the globe are facing catastrophic declines, in part because of infections with pathogens such as the frog virus 3 (fv3) ranavirus; the mechanisms governing amphibian susceptibility and resistance to such pathogens remain poorly understood. the type i and type iii interferon (ifn) cytokines represent a cornerstone of vertebrate antiviral immunity, while our recent work indicates that tadpoles and adult frogs of the amphibian xenopus laevis may differ in their ifn responses ...201830018186
the b-hlh transcription factor hes3 participates in neural plate border formation by interfering with wnt/β-catenin signaling.hes3 belongs to the hes basic helix-loop-helix family of transcriptional repressors that play central roles in maintaining progenitor cells and regulating binary cell fate decisions in the embryo. during xenopus laevis development, hes3 is expressed in the embryonic ectoderm in a horseshoe shape domain at the edge of the developing neural pate. hes3 mis-expression at early neurula stage blocks neural crest (snai2, sox8, sox9 and sox10) and cranial placode (six1 and dmrta1) gene expression, and p ...201830016640
plc and ip3-evoked ca2+ release initiate the fast block to polyspermy in xenopus laevis eggs.the prevention of polyspermy is essential for the successful progression of normal embryonic development in most sexually reproducing species. in external fertilizers, the process of fertilization induces a depolarization of the egg's membrane within seconds, which inhibits supernumerary sperm from entering an already-fertilized egg. this fast block requires an increase of intracellular ca2+ in the african clawed frog, xenopus laevis, which in turn activates an efflux of cl- that depolarizes the ...201830012841
comprehensive substrate characterization of 22 antituberculosis drugs for multiple solute carrier (slc) uptake transporters in vitro.the substrate potentials of antituberculosis drugs on solute carrier (slc) transporters are not well characterized to date, despite a well-established understanding of their drug dispositions and pharmacokinetics. in this study, we investigated comprehensively the substrate potentials of the 22 currently available antituberculosis drugs for slc family transporter-mediated uptake, using xenopus laevis oocytes and stably transfected hek-293 cells in vitro the result suggested that ethambutol, ison ...201830012768
a gene regulatory network underlying the formation of pre-placodal ectoderm in xenopus laevis.the neural plate border ectoderm gives rise to key developmental structures during embryogenesis, including the neural crest and the preplacodal ectoderm. many sensory organs and ganglia of vertebrates develop from cranial placodes, which themselves arise from preplacodal ectoderm, defined by expression of transcription factor six1 and its coactivator eya1. here we elucidate the gene regulatory network underlying the specification of the preplacodal ectoderm in xenopus, and the functional intera ...201830012125
water contaminants associated with unconventional oil and gas extraction cause immunotoxicity to amphibian tadpoles.chemicals associated with unconventional oil and gas (uog) operations have been shown to contaminate surface and ground water with a variety of endocrine disrupting compounds (edcs) inducing multiple developmental alteration in mice. however, little is known about the impacts of uog-associated contaminants on amphibian health and resistance to an emerging ranavirus infectious disease caused by viruses in the genus ranavirus, especially at the vulnerable tadpole stage. here we used tadpoles of th ...201830011011
thiodipeptides targeting the intestinal oligopeptide transporter as a general approach to improving oral drug delivery.the broad substrate capacity of the intestinal oligopeptide transporter, pept1, has made it a key target of research into drug delivery. whilst the substrate capacity of this transporter is broad, studies have largely been limited to small peptides and peptide-like drugs. here, we demonstrate for the first time that a diverse range of drugs can be targeted towards transport by pept1 using a hydrolysis resistant carrier. eleven prodrugs were synthesized by conjugating modified dipeptides containi ...201830006163
pharmacological and toxicological activity of rsd921, a novel sodium channel blocker.rsd921, the r,r enantiomer of the kappa (k) agonist pd117,302, lacks significant activity on opioid receptors.201829990839
human mutations in slc2a9 (glut9) affect transport capacity for urate.slc2a9 or glut9 is a voltage sensitive urate transporter, mainly expressed in the kidneys, the liver, and the intestine. human glut9 loss-of-function mutations were identified in familial hypouricemia, and several single nucleotide polymorphisms (snps) were associated with lower serum urate, further indicating that glut9 is a major determinant of serum uric acid level. to get insights in glut9 transport characteristics, we systematically analyzed the function of known human glut9 mutants using 1 ...201829967582
the fungal alkaloid okaramine-b activates an l-glutamate-gated chloride channel from ixodes scapularis, a tick vector of lyme disease.a novel l-glutamate-gated anion channel (iscaglucl1) has been cloned from the black-legged tick, ixodes scapularis, which transmits multiple pathogens including the agents of lyme disease and human granulocytic anaplasmosis. when mrna encoding iscaglucl1 was expressed in xenopus laevis oocytes, we detected robust 50-400 na currents in response to 100 μm l-glutamate. responses to l-glutamate were concentration-dependent (pec50 3.64 ± 0.11). ibotenate was a partial agonist on iscaglucl1. we detect ...201829957333
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
influenza d virus m2 protein exhibits ion channel activity in xenopus laevis oocytes.a new type of influenza virus, known as type d, has recently been identified in cattle and pigs. influenza d virus infection in cattle is typically asymptomatic; however, its infection in swine can result in clinical disease. swine can also be infected with all other types of influenza viruses, namely a, b, and c. consequently, swine can serve as a "mixing vessel" for highly pathogenic influenza viruses, including those with zoonotic potential. currently, the only antiviral drug available target ...201829927982
probing proline residues in the prokaryotic ligand-gated ion channel, elic.erwinia ligand-gated ion channel (elic) is a bacterial homologue of vertebrate pentameric ligand-gated ion channels (plgics) and has proven to be a valuable model for understanding the structure and function of this important protein family. there is nevertheless still a question about whether molecular details can be accurately extrapolated from this protein to those found in eukaryotes. here we explore the role of proline residues (pros) in elic by creating mutant receptors, expressing them in ...201829927250
nuclear arp2/3 drives dna break clustering for homology-directed repair.dna double-strand breaks repaired by non-homologous end joining display limited dna end-processing and chromosomal mobility. by contrast, double-strand breaks undergoing homology-directed repair exhibit extensive processing and enhanced motion. the molecular basis of this movement is unknown. here, using xenopus laevis cell-free extracts and mammalian cells, we establish that nuclear actin, wasp, and the actin-nucleating arp2/3 complex are recruited to damaged chromatin undergoing homology-direc ...201829925947
gitelman's syndrome: characterization of a novel c.1181g>a point mutation and functional classification of the known mutations.we have investigated the mechanisms by which a novel missense point mutation (c.1181g>a) found in two sisters causes gitelman's syndrome by impairing the sodium chloride co-transporter (ncc, encoded by slc12a3 gene) function. the cdna and in vitro transcribed mrna of either wild-type or mutated slc12a3 were transfected into hek293 cells and injected into xenopus laevis oocytes, respectively. the expression, maturation, trafficking, and function of the mutated and wild-type ncc were assessed by w ...201829925901
regulation of nuclear factor of activated t cells (nfat) and downstream myogenic proteins during dehydration in the african clawed frog.xenopus laevis, otherwise known as the african clawed frog, undergoes natural dehydration of up to 30% of its total body water during the dry season in sub-saharan africa. to survive under these conditions, a variety of physiological and biochemical changes take place in x. laevis. we were interested in understanding the role that the calcineurin-nfat pathway plays during dehydration stress response in the skeletal muscles of x. laevis. immunoblotting was performed to characterize the protein le ...201829923155
semicircular canal influences on the developmental tuning of the translational vestibulo-ocular reflex.vestibulo-ocular reflexes (vors) rely on neuronal computations that transform vestibular sensory signals into spatio-temporally appropriate extraocular motor commands. the motoneuronal discharge for contractions of the superior oblique eye muscle during linear translation derives from a utricular epithelial sector that is spatially aligned with the pulling direction of this muscle. in xenopus laevis, the alignment is gradually achieved during larval development and requires motion-related semici ...201829922219
a wide variety of mitf transcript variants are expressed in the xenopus laevis periodic albino mutant.the periodic albino mutant of xenopus laevis has been used to study the development of pigment cells because both the retinal pigment epithelium (rpe) and melanophores are affected. in this mutant, "white pigment cells" containing both melanophore-specific and iridophore-specific pigment organelles appear. the present experiments were designed to investigate the structural organization and expression of microphthalmia-associated transcription factor (mitf) in the mutant since mitf is known to re ...201829920861
a sodium transporter hvhkt1;1 confers salt tolerance in barley via regulating tissue and cell ion homeostasis.our previous studies showed that high salt tolerance in tibetan wild barley accessions was associated with hvhkt1;1, a member of the high-affinity potassium transporter family. however, molecular mechanisms of hvhkt1;1 for salt tolerance and its roles in k+/na+ homeostasis remain to be elucidated. functional characterization of hvhkt1;1 was conducted in the present study. nacl-induced transcripts of hvhkt1;1 were significantly higher in the roots of tibetan wild barley xz16 relative to other gen ...201829917153
isolation and functional characterization of cslsi2, a cucumber silicon efflux transporter gene.silicon has been proven to exert beneficial effects on plant growth and stress tolerance, and silicon accumulation varies among different plant species. cucumber (cucumis sativus) is a widely used dicot model for silicon accumulation, but little is known about the molecular mechanism of its silicon uptake. previously, we isolated and characterized cslsi1, a silicon influx transporter gene from cucumber. in this study, we cloned a putative silicon efflux transporter gene, cslsi2, from cucumber an ...201829905780
the rna-binding complex escrt-ii in xenopus laevis eggs recognizes purine-rich sequences through its subunit, vps25.rna-binding proteins (rbp) are critical regulators of gene expression. recent studies have uncovered hundreds of mrna-binding proteins that do not contain annotated rna-binding domains and have well-established roles in other cellular processes. investigation of these nonconventional rbps is critical for revealing novel rna-binding domains and may disclose connections between rna regulation and other aspects of cell biology. the endosomal sorting complex required for transport ii (escrt-ii) is a ...201829903915
transcriptome of xenopus andrei, an octoploid frog, during embryonic development.although polyploidy occurs throughout the fish and amphibian lineages, the xenopus genus exhibits a high incidence of polyploidy, with 25 out of the 26 known species being polyploid. however, transcriptomic information is currently available for only one of these species, the tetraploid xenopus laevis. xenopus andrei, an octoploid species within the xenopus genus, offers an opportunity for assessing a novel polyploid transcriptome during vertebrate development. rna-seq data was generated at nine ...201829900348
validation and bioinformatics analysis of differentially expressed circrnas involved in developing male xenopus laevis chronically exposed to atrazine.the data presented here are related to the research article titled "identification of circular rnas and their alterations involved in developing male xenopus laevis chronically exposed to atrazine" (sai et al., 2018) [1]. circular rnas (circrnas) are implicated in multiple developmental anomalies (bachmayr-heyda et al., 2015; li et al., 2015) [2], [3]. this report describes the differentially expressed circrnas involved in developing male xenopus laevis (x. laevis) chronically exposed to atrazin ...201829900306
visualization of gene expression patterns by in situ hybridization on early stages of development of xenopus laevis.in situ hybridization performed using whole fixed embryos provides accurate and detailed visualization of gene expression patterns. these patterns are useful for investigating spatial patterns of gene expression in normally developing embryos but can also be useful in investigating the effects of genetic or environmental changes on expression of genetic markers characteristic of particular tissues, organs, or genetic pathways. our lab's protocol for whole-mount in situ hybridization is presented ...201829896701
evaluation of teratogenicity of pharmaceuticals using fetax.pharmaceuticals are chemical compounds which are used to preserve human and animal health. once administered, these compounds are metabolized or can remain unaltered until excreted. therefore, a mixture of pharmaceuticals and their metabolites enters municipal sewers and wastewater treatment plants where, depending on their polarity, water solubility, and persistence, they cannot be completely removed or transformed during the treatment process, so that unaltered pharmaceuticals and/or their met ...201829896699
inward- and outward-facing homology modeling of human concentrative nucleoside transporter 3 (hcnt3) predicts an elevator-type transport mechanism.the human slc28 family of concentrative (na+-dependent) nucleoside transporters has three members, hcnt1, hcnt2 and hcnt3. previously, we have used heterologous expression in xenopus laevis oocytes in combination with an engineered cysteine-less hcnt3 protein hcnt3(c-) to undertake systematic substituted cysteine accessibility method (scam) analysis of the transporter using the membrane-impermeant thiol reactive reagent p-chloromercuribenzene sulfonate (pcmbs). a continuous sequence of more than ...201830096006
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
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
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
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
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
nucleosomes and the three glycosylases: high, medium, and low levels of excision by the uracil dna glycosylase superfamily.human cells express the udg superfamily of glycosylases, which excise uracil (u) from the genome. the three members of this structural superfamily are uracil dna glycosylase (ung/udg), single-strand selective monofunctional uracil dna glycosylase (smug1), and thymine dna glycosylase (tdg). we previously reported that udg is efficient at removing u from dna packaged into nucleosome core particles (ncp) and is minimally affected by the histone proteins when acting on an outward-facing u in the dya ...201830268365
a xenopus oocyte model system to study action potentials.action potentials (aps) are the functional units of fast electrical signaling in excitable cells. the upstroke and downstroke of an ap is generated by the competing and asynchronous action of na+- and k+-selective voltage-gated conductances. although a mixture of voltage-gated channels has been long recognized to contribute to the generation and temporal characteristics of the ap, understanding how each of these proteins function and are regulated during electrical signaling remains the subject ...201830266757
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
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
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
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
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
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
developmental dependence for functional eye regrowth in xenopus laevis. 201830136686
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
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
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
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