36 results on '"John B. Wallingford"'
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2. Spatiotemporal transcriptional dynamics of the cycling mouse oviduct
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Elle C. Roberson, Anna Battenhouse, Edward M. Marcotte, Ngan Kim Tran, John B. Wallingford, and Riddhiman K. Garge
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endocrine system ,Embryonic Development ,Gene Expression ,Estrous Cycle ,Oviducts ,Biology ,Article ,Transcriptome ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Pregnancy ,Gene expression ,Genetic model ,Animals ,Hox gene ,Molecular Biology ,Gene ,reproductive and urinary physiology ,030304 developmental biology ,Estrous cycle ,0303 health sciences ,urogenital system ,Gene Expression Profiling ,Dynamics (mechanics) ,Embryogenesis ,Cell Biology ,Cell biology ,Fertility ,Gene Expression Regulation ,Oviduct ,Female ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Female fertility in mammals requires iterative remodeling of the entire adult female reproductive tract across the menstrual/estrous cycle. However, while transcriptome dynamics across the estrous cycle have been reported in human and bovine models, no global analysis of gene expression across the estrous cycle has yet been reported for the mouse. Here, we examined the cellular composition and global transcriptional dynamics of the mouse oviduct along the anteroposterior axis and across the estrous cycle. We observed robust patterns of differential gene expression along the anteroposterior axis, but we found surprisingly few changes in gene expression across the estrous cycle. Notable gene expression differences along the anteroposterior axis included a surprising enrichment for genes related to embryonic development, such as Hox and Wnt genes. The relatively stable transcriptional dynamics across the estrous cycle differ markedly from other mammals, leading us to speculate that this is an evolutionarily derived state that may reflect the extremely rapid five-day mouse estrous cycle. This dataset fills a critical gap by providing an important genomic resource for a highly tractable genetic model of mammalian female reproduction. more...
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- 2021
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3. A systematic, label-free method for identifying RNA-associated proteins in vivo provides insights into vertebrate ciliary beating machinery
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Ryan L. Huizar, Vy Dang, John B. Wallingford, Ophelia Papoulas, Edward M. Marcotte, Chanjae Lee, Caitlin C. Devitt, Kevin Drew, Claire D. McWhite, and Rachael M. Cox
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Embryo, Nonmammalian ,Xenopus ,Xenopus Proteins ,Biology ,Proteomics ,Epithelium ,Article ,Tissue Culture Techniques ,03 medical and health sciences ,0302 clinical medicine ,Animals ,Cilia ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Cilium ,RNA-Binding Proteins ,RNA ,Cell Biology ,Embryonic Tissue ,Inner dynein arm ,Cell biology ,Spindle apparatus ,Cytoplasm ,Proteome ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Cell-type specific RNA-associated proteins are essential for development and homeostasis in animals. Despite a massive recent effort to systematically identify RNA-associated proteins, we currently have few comprehensive rosters of cell-type specific RNA-associated proteins in vertebrate tissues. Here, we demonstrate the feasibility of determining the RNA-interacting proteome of a defined vertebrate embryonic tissue using DIF-FRAC, a systematic and universal (i.e., label-free) method. Application of DIF-FRAC to cultured tissue explants of Xenopus mucociliary epithelium identified dozens of known RNA-associated proteins as expected, but also several novel RNA-associated proteins, including proteins related to assembly of the mitotic spindle and regulation of ciliary beating. In particular, we show that the inner dynein arm tether Cfap44 is an RNA-associated protein that localizes not only to axonemes, but also to liquid-like organelles in the cytoplasm called DynAPs. This result led us to discover that DynAPs are generally enriched for RNA. Together, these data provide a useful resource for a deeper understanding of mucociliary epithelia and demonstrate that DIF-FRAC will be broadly applicable for systematic identification of RNA-associated proteins from embryonic tissues. more...
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- 2020
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4. PCP and Septins Govern the Polarized Organization and Mechanics of the Actin Cytoskeleton During Convergent Extension
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Caitlin C Devitt, Shinuo Weng, Vidal D Bejar-Padilla, José Alvarado, and John B Wallingford
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SummaryConvergent extension (CE) is a collective cell movement required for embryonic axis elongation, neural tube closure, and kidney tubule elongation. During CE, iterative cell intercalations drive the elongation of a tissue 1. In order to move, cells reorganize their actin cytoskeleton; the implication of actin regulators during CE, then, is expected. Interestingly, two other groups of proteins have been shown to be required for this cell behavior: Planar Cell Polarity proteins (PCP)2-4 and Septins5, both of which interact with the actin cytoskeleton 6-14. Disruption of any of these systems have substantial overlap in terms of embryonic phenotypes, but how they interact to govern CE is largely unknown15-17. Here, we find that PCP proteins and Septins sequester actomyosin activity, create flows of actin across the anterior side of cells, and bundle actin into a node and cable system that displays a planar polarized gradient of stiffness.Key findings-PCP- and Septin-dependent actin node and cable structures form and become anteriorized over time-Actin cytoskeletal reorganization correlates with PCP protein polarization.-Actomyosin flows are directed to anterior PCP puncta and PCP movement relies on actin turnover.-Septins localize to anterior nodes and bundle actin filaments into anterior actin cable.-PCP- and Septin-dependent anterior actin cables increase local cellular stiffness and create a planar polarized stiffness gradient across the cell. more...
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- 2022
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5. We Are All Developmental Biologists
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John B. Wallingford
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Cognitive science ,0303 health sciences ,Stem Cells ,Genetic Diseases, Inborn ,Face (sociological concept) ,Cell Biology ,Biology ,Embryo, Mammalian ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,0302 clinical medicine ,Transformative learning ,Humans ,Regeneration ,Regeneration (ecology) ,Molecular Biology ,Developmental biology ,030217 neurology & neurosurgery ,030304 developmental biology ,Developmental Biology - Abstract
Humans have sought to understand the embryo for millennia. Paradoxically, even as technical and intellectual innovations bring us ever closer to a transformative understanding of developmental biology, our discipline faces an "image problem." We should face this problem by acknowledging that developmental biology is fundamental to the human experience. more...
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- 2019
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6. Commentary and tribute to Antone Jacobson: The pioneer of morphodynamics
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John B. Wallingford and Amy K. Sater
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Embryology ,Teaching ,Art history ,Tribute ,Historical Article ,Biography ,Cell Biology ,History, 20th Century ,Biology ,Salamandridae ,History, 21st Century ,United States ,Morphogenesis ,Animals ,Drosophila ,Molecular Biology ,Beach morphodynamics ,Developmental Biology - Published
- 2019
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7. Cell Adhesions Link Subcellular Actomyosin Dynamics to Tissue Scale Force Production During Vertebrate Convergent Extension
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Chaehee Lee, Sena Sarıkaya, John B. Wallingford, Ophelia Papoulas, Rachael M. Cox, Shinuo Weng, Robert J. Huebner, and Edward M. Marcotte
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Multicellular organism ,Convergent extension ,Catenin ,Armadillo repeats ,Biology ,Cell adhesion ,Cytoskeleton ,Proteomics ,Phenotype ,Cell biology - Abstract
SummaryAxis extension is a fundamental biological process that shapes multicellular organisms. The design of an animal’s body plan is encoded in the genome and execution of this program is a multiscale mechanical progression involving the coordinated movement of proteins, cells, and whole tissues. Thus, a key challenge to understanding axis extension is connecting events that occur across these various length scales. Here, we use approaches from proteomics, cell biology, and tissue biomechanics to describe how a poorly characterized cell adhesion effector, the Armadillo Repeat protein deleted in Velo-Cardio-Facial syndrome (Arvcf) catenin, controls vertebrate head-to-tail axis extension. We find that Arvcf catenin is required for axis extension within the intact organism but is not required for extension of isolated tissues. We then show that the organism scale phenotype is caused by a modest defect in force production at the tissue scale that becomes apparent when the tissue is challenged by external resistance. Finally, we show that the tissue scale force defect results from dampening of the pulsatile recruitment of cell adhesion and cytoskeletal proteins to cell membranes. These results not only provide a comprehensive understanding of Arvcf function during an essential biological process, but also provide insight into how a modest cellular scale defect in cell adhesion results in an organism scale failure of development. more...
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- 2021
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8. PCP-dependent transcellular regulation of actomyosin oscillation facilitates convergent extension of vertebrate tissue
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Asako Shindo, John B. Wallingford, Yasuhiro Inoue, and Makoto Kinoshita
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Embryo, Nonmammalian ,Contraction (grammar) ,Central nervous system ,Cell ,Xenopus Proteins ,Models, Biological ,Time-Lapse Imaging ,Article ,Xenopus laevis ,03 medical and health sciences ,0302 clinical medicine ,Cell Movement ,biology.animal ,medicine ,Animals ,Transcellular ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Microscopy, Confocal ,biology ,Convergent extension ,Oscillation ,Cell Membrane ,Neural tube ,Cell Polarity ,Gene Expression Regulation, Developmental ,Vertebrate ,Actomyosin ,Cell Biology ,Cell biology ,Luminescent Proteins ,medicine.anatomical_structure ,Algorithms ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Oscillatory flows of actomyosin play a key role in the migration of single cells in culture and in collective cell movements in Drosophila embryos. In vertebrate embryos undergoing convergent extension (CE), the Planar Cell Polarity (PCP) pathway drives the elongation of the body axis and shapes the central nervous system, and mutations of the PCP genes predispose humans to various malformations including neural tube defects. However, the spatiotemporal patterns of oscillatory actomyosin contractions during vertebrate CE and how they are controlled by the PCP signaling remain unknown. Here, we address these outstanding issues using a combination of in vivo imaging and mathematical modeling. We find that effective execution of CE requires alternative oscillations of cortical actomyosin across cell membranes of neighboring cells within an optimal frequency range. Intriguingly, temporal and spatial clustering of the core PCP protein Prickle 2 (Pk2) is correlated to submembranous accumulations of F-actin, and depletion of Pk2 perturbs the oscillation of actomyosin contractions. These findings shed light on the significance of temporal regulation of actomyosin contraction by the PCP pathway during CE, in addition to its well-studied spatial aspects. more...
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- 2019
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9. ARVCF catenin controls force production during vertebrate convergent extension
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Robert J, Huebner, Shinuo, Weng, Chanjae, Lee, Sena, Sarıkaya, Ophelia, Papoulas, Rachael M, Cox, Edward M, Marcotte, and John B, Wallingford
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Armadillo Domain Proteins ,Xenopus laevis ,Morphogenesis ,Animals ,Catenins ,Cell Biology ,Cadherins ,Phosphoproteins ,Cell Adhesion Molecules ,Molecular Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,Developmental Biology - Abstract
The design of an animal's body plan is encoded in the genome, and the execution of this program is a mechanical progression involving coordinated movement of proteins, cells, and whole tissues. Thus, a challenge to understanding morphogenesis is connecting events that occur across various length scales. Here, we describe how a poorly characterized adhesion effector, Arvcf catenin, controls Xenopus head-to-tail axis extension. We find that Arvcf is required for axis extension within the intact organism but not within isolated tissues. We show that the organism-scale phenotype results from a defect in tissue-scale force production. Finally, we determine that the force defect results from the dampening of the pulsatile recruitment of cell adhesion and cytoskeletal proteins to membranes. These results provide a comprehensive understanding of Arvcf function during axis extension and produce an insight into how a cellular-scale defect in adhesion results in an organism-scale failure of development. more...
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- 2022
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10. Convergent extension requires adhesion-dependent biomechanical integration of cell crawling and junction contraction
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Robert J. Huebner, John B. Wallingford, and Shinuo Weng
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Physics ,Convergent extension ,Cell ,Morphogenesis ,Neural tube ,Crawling ,General Biochemistry, Genetics and Molecular Biology ,medicine.anatomical_structure ,Live cell imaging ,Cell Movement ,Catenin ,medicine ,Cell Adhesion ,Humans ,Neural Tube Defects ,Cell adhesion ,Neuroscience ,Cell Adhesion Molecules - Abstract
Convergent extension is an evolutionarily conserved collective cell movement that elongates the body axis of almost all animals and is required for the morphogenesis of several organ systems. Decades of study have revealed two distinct mechanisms of cell movement during CE, one based on cell crawling and the other on junction contraction. How these two behaviors collaborate during CE is not understood. Here, using quantitative live cell imaging we show that these two modes act both independently and in concert during CE, but that cell movement is more effective when the two modes are integrated via mechano-reciprocity. Based on these findings, we developed a novel computational model that for the first time treats crawling and contraction independently. This model not only confirmed the biomechanical efficacy of integrating the two modes, but also revealed for the first time how the two modes -and their integration- are influenced by cell adhesion. Finally, we use these new insights to further understand the complex CE phenotype resulting from loss of the C-cadherin interacting catenin Arvcf. These data are significant for providing new biomechanical and cell biological insights into a fundamental morphogenetic process that is implicated in human neural tube defects and skeletal dysplasias. more...
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- 2022
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11. Identification of new regulators of embryonic patterning and morphogenesis in Xenopus gastrulae by RNA sequencing
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Chenbei Chang, Taejoon Kwon, Ivan K. Popov, David K. Crossman, John B. Wallingford, and Michael R. Crowley
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0301 basic medicine ,Embryo, Nonmammalian ,Polarity in embryogenesis ,Xenopus ,Morphogenesis ,Xenopus Proteins ,Cell fate determination ,Article ,03 medical and health sciences ,Animals ,RNA, Messenger ,Molecular Biology ,Body Patterning ,Genetics ,Extracellular Matrix Proteins ,biology ,Sequence Analysis, RNA ,Convergent extension ,Organizers, Embryonic ,Embryogenesis ,Gene Expression Regulation, Developmental ,Gastrula ,Cell Biology ,Protein-Tyrosine Kinases ,biology.organism_classification ,Actin cytoskeleton ,Activins ,Cell biology ,Gastrulation ,030104 developmental biology ,embryonic structures ,Germ Layers ,Developmental Biology - Abstract
During early vertebrate embryogenesis, cell fate specification is often coupled with cell acquisition of specific adhesive, polar and/or motile behaviors. In Xenopus gastrulae, tissues fated to form different axial structures display distinct motility. The cells in the early organizer move collectively and directionally toward the animal pole and contribute to anterior mesendoderm, whereas the dorsal and the ventral-posterior trunk tissues surrounding the blastopore of mid-gastrula embryos undergo convergent extension and convergent thickening movements, respectively. While factors regulating cell lineage specification have been described in some detail, the molecular machinery that controls cell motility is not understood in depth. To gain insight into the gene battery that regulates both cell fates and motility in particular embryonic tissues, we performed RNA sequencing (RNA-seq) to investigate differentially expressed genes in the early organizer, the dorsal and the ventral marginal zone of Xenopus gastrulae. We uncovered many known signaling and transcription factors that have been reported to play roles in embryonic patterning during gastrulation. We also identified many uncharacterized genes as well as genes that encoded extracellular matrix (ECM) proteins or potential regulators of actin cytoskeleton. Co-expression of a selected subset of the differentially expressed genes with activin in animal caps revealed that they had distinct ability to block activin-induced animal cap elongation. Most of these factors did not interfere with mesodermal induction by activin, but an ECM protein, EFEMP2, inhibited activin signaling and acted downstream of the activated type I receptor. By focusing on a secreted protein kinase PKDCC1, we showed with overexpression and knockdown experiments that PKDCC1 regulated gastrulation movements as well as anterior neural patterning during early Xenopus development. Overall, our studies identify many differentially expressed signaling and cytoskeleton regulators in different embryonic regions of Xenopus gastrulae and imply their functions in regulating cell fates and/or behaviors during gastrulation. more...
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- 2017
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12. From Planar Cell Polarity to Ciliogenesis and Back: The Curious Tale of the PPE and CPLANE proteins
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Paul N. Adler and John B. Wallingford
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0301 basic medicine ,chemistry.chemical_classification ,Frizzled ,Effector ,Organogenesis ,Cilium ,Cell Polarity ,Membrane Proteins ,Cell Biology ,Biology ,Actin cytoskeleton ,Article ,Dishevelled ,Cell biology ,Actin Cytoskeleton ,03 medical and health sciences ,030104 developmental biology ,chemistry ,Membrane protein ,Ciliogenesis ,Cell polarity ,Animals ,Humans ,Disease ,Cilia - Abstract
Why some genes are more popular than others remains an open question, but one example of this phenomenon involves the genes controlling planar cell polarity (PCP), the polarization of cells within a plane of a tissue. Indeed, the so-called 'core' PCP genes such as dishevelled, frizzled, and prickle have been extensively studied both in animal models and by human genetics. By contrast, other genes that influence PCP signaling have received far less attention. Among the latter are inturned, fuzzy, and fritz, but recent work should bring these once obscure regulators into the limelight. We provide here a brief history of planar polarity effector (PPE) and CPLANE (ciliogenesis and planar polarity effector) proteins, discuss recent advances in understanding their molecular mechanisms of action, and describe their roles in human disease. more...
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- 2017
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13. A novel ciliopathic skull defect arising from excess neural crest
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Karen J. Liu, John B. Wallingford, Jacqueline M. Tabler, and Christopher P. Rice
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0301 basic medicine ,Greig cephalopolysyndactyly ,Gene Dosage ,Ciliopathies ,Fgf8 ,Craniofacial Abnormalities ,Mesoderm ,Mice ,Morphogenesis ,Intracellular Signaling Peptides and Proteins ,Gene Expression Regulation, Developmental ,Neural crest ,Anatomy ,craniosynostosis ,medicine.anatomical_structure ,Neural Crest ,coronal suture ,neural crest ,Signal Transduction ,skull ,calvaria ,Fibroblast Growth Factor 8 ,Wnt1 ,Kruppel-Like Transcription Factors ,morphogenesis ,Mesp1 ,Mice, Transgenic ,Nerve Tissue Proteins ,Calvaria ,Biology ,craniofacial ,Article ,Craniosynostosis ,Parietal Bone ,03 medical and health sciences ,Zinc Finger Protein Gli3 ,medicine ,Animals ,Cilia ,Molecular Biology ,mouse ,Fuz ,Skull ,Cell Biology ,medicine.disease ,Cytoskeletal Proteins ,Ciliopathy ,ciliopathy ,030104 developmental biology ,Frontal bone ,Frontal Bone ,Coronal suture ,Developmental Biology - Abstract
The skull is essential for protecting the brain from damage, and birth defects involving disorganization of skull bones are common. However, the developmental trajectories and molecular etiologies by which many craniofacial phenotypes arise remain poorly understood. Here, we report a novel skull defect in ciliopathic Fuz mutant mice in which only a single bone pair encases the forebrain, instead of the usual paired frontal and parietal bones. Through genetic lineage analysis, we show that this defect stems from a massive expansion of the neural crest-derived frontal bone. This expansion occurs at the expense of the mesodermally-derived parietal bones, which are either severely reduced or absent. A similar, though less severe, phenotype was observed in Gli3 mutant mice, consistent with a role for Gli3 in cilia-mediated signaling. Excess crest has also been shown to drive defective palate morphogenesis in ciliopathic mice, and that defect is ameliorated by reduction of fgf8 gene dosage. Strikingly, skull defects in Fuz mutant mice are also rescued by loss of one allele of fgf8, suggesting a potential route to therapy. In sum, this work is significant for revealing a novel skull defect with a previously un-described developmental etiology and for suggesting a common developmental origin for skull and palate defects in ciliopathies. more...
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- 2016
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14. TTC25 Deficiency Results in Defects of the Outer Dynein Arm Docking Machinery and Primary Ciliary Dyskinesia with Left-Right Body Asymmetry Randomization
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Heymut Omran, Gerard W. Dougherty, Julia Wallmeier, John B. Wallingford, Heike Olbrich, Yasuko Asai, Chanjae Lee, Sascha Sauer, Yasin Memari, Kyosuke Shinohara, Petra Pennekamp, Matthias Griese, Johanna Raidt, Anja Kolb-Kokocinski, Tabea Menchen, Hidetaka Shiratori, Rim Hjeij, Christine Edelbusch, Richard Durbin, Claudius Werner, Hiroshi Hamada, Katsura Minegishi, Niki T. Loges, and Katsuyoshi Takaoka more...
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0301 basic medicine ,Axoneme ,Xenopus ,Dynein ,Fluorescent Antibody Technique ,Genes, Recessive ,Biology ,Xenopus Proteins ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Microscopy, Electron, Transmission ,Report ,medicine ,Genetics ,Animals ,Humans ,Exome ,Genetics(clinical) ,Cilia ,Genetics (clinical) ,Primary ciliary dyskinesia ,Kartagener Syndrome ,Cilium ,Dyneins ,Anatomy ,Exons ,medicine.disease ,biology.organism_classification ,Cell biology ,030104 developmental biology ,030220 oncology & carcinogenesis ,Mutation ,Outer dynein arm ,NODAL ,Carrier Proteins ,Congenital disorder ,Protein Binding - Abstract
Multiprotein complexes referred to as outer dynein arms (ODAs) develop the main mechanical force to generate the ciliary and flagellar beat. ODA defects are the most common cause of primary ciliary dyskinesia (PCD), a congenital disorder of ciliary beating, characterized by recurrent infections of the upper and lower airways, as well as by progressive lung failure and randomization of left-right body asymmetry. Using a whole-exome sequencing approach, we identified recessive loss-of-function mutations within TTC25 in three individuals from two unrelated families affected by PCD. Mice generated by CRISPR/Cas9 technology and carrying a deletion of exons 2 and 3 in Ttc25 presented with laterality defects. Consistently, we observed immotile nodal cilia and missing leftward flow via particle image velocimetry. Furthermore, transmission electron microscopy (TEM) analysis in TTC25-deficient mice revealed an absence of ODAs. Consistent with our findings in mice, we were able to show loss of the ciliary ODAs in humans via TEM and immunofluorescence (IF) analyses. Additionally, IF analyses revealed an absence of the ODA docking complex (ODA-DC), along with its known components CCDC114, CCDC151, and ARMC4. Co-immunoprecipitation revealed interaction between the ODA-DC component CCDC114 and TTC25. Thus, here we report TTC25 as a new member of the ODA-DC machinery in humans and mice. more...
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- 2016
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15. Proteome-wide dataset supporting the study of ancient metazoan macromolecular complexes
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Zuyao Ni, Mihail Sarov, Pierre C. Havugimana, Edward M. Marcotte, Julian Kwan, Sadhna Phanse, Ramy H. Malty, Kyle Chessman, Ophelia Papoulas, Xuejian Xiong, Xinghua Guo, Blake Borgeson, Jack Greenblatt, Mohan Babu, John B. Wallingford, John Parkinson, W. Brent Derry, Snejana Stoilova, Alexandr Bezginov, Elisabeth R. M. Tillier, Swati Pal, Daniel R. Boutz, Kevin Drew, Greg W. Clark, Graham L. Cromar, Olga Kagan, Cuihong Wan, Andrew Emili, and Fan Tu more...
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0301 basic medicine ,Proteomics ,Protein complexes ,Computational biology ,Bioinformatics ,lcsh:Computer applications to medicine. Medical informatics ,Dictyostelium discoideum ,Biochemical ,03 medical and health sciences ,Interaction network ,Fractionation ,lcsh:Science (General) ,Caenorhabditis elegans ,Data Article ,Multidisciplinary ,biology ,Metazoa ,biology.organism_classification ,Strongylocentrotus purpuratus ,030104 developmental biology ,Homo sapiens ,Proteome ,lcsh:R858-859.7 ,Drosophila melanogaster ,lcsh:Q1-390 - Abstract
Our analysis examines the conservation of multiprotein complexes among metazoa through use of high resolution biochemical fractionation and precision mass spectrometry applied to soluble cell extracts from 5 representative model organisms Caenorhabditis elegans, Drosophila melanogaster, Mus musculus, Strongylocentrotus purpuratus, and Homo sapiens. The interaction network obtained from the data was validated globally in 4 distant species (Xenopus laevis, Nematostella vectensis, Dictyostelium discoideum, Saccharomyces cerevisiae) and locally by targeted affinity-purification experiments. Here we provide details of our massive set of supporting biochemical fractionation data available via ProteomeXchange (http://www.ebi.ac.uk/pride/archive/projects/PXD002319-http://www.ebi.ac.uk/pride/archive/projects/PXD002328), PPIs via BioGRID (185267); and interaction network projections via (http://metazoa.med.utoronto.ca) made fully accessible to allow further exploration. The datasets here are related to the research article on metazoan macromolecular complexes in Nature [1]. Keywords: Proteomics, Metazoa, Protein complexes, Biochemical, Fractionation more...
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- 2016
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16. Emergence of an Apical Epithelial Cell Surface In Vivo
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Maté Biro, Jakub Sedzinski, Fan Tu, John B. Wallingford, and Edouard Hannezo
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0301 basic medicine ,Xenopus ,Cell ,Cell Separation ,Article ,Epithelium ,General Biochemistry, Genetics and Molecular Biology ,Tight Junctions ,03 medical and health sciences ,medicine ,Animals ,Homeostasis ,Molecular Biology ,Process (anatomy) ,Actin ,Epithelial polarity ,biology ,Tight junction ,Epithelial Cells ,Cell Biology ,biology.organism_classification ,Actins ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Developmental Biology - Abstract
SummaryEpithelial sheets are crucial components of all metazoan animals, enclosing organs and protecting the animal from its environment. Epithelial homeostasis poses unique challenges, as addition of new cells and loss of old cells must be achieved without disrupting the fluid-tight barrier and apicobasal polarity of the epithelium. Several studies have identified cell biological mechanisms underlying extrusion of cells from epithelia, but far less is known of the converse mechanism by which new cells are added. Here, we combine molecular, pharmacological, and laser-dissection experiments with theoretical modeling to characterize forces driving emergence of an apical surface as single nascent cells are added to a vertebrate epithelium in vivo. We find that this process involves the interplay between cell-autonomous actin-generated pushing forces in the emerging cell and mechanical properties of neighboring cells. Our findings define the forces driving this cell behavior, contributing to a more comprehensive understanding of epithelial homeostasis. more...
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- 2016
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17. Zeta-Tubulin Is a Member of a Conserved Tubulin Module and Is a Component of the Centriolar Basal Foot in Multiciliated Cells
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Airon A. Wills, Tim Stearns, Erin Turk, Taejoon Kwon, John B. Wallingford, and Jakub Sedzinski
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Centriole ,Agricultural and Biological Sciences(all) ,Biochemistry, Genetics and Molecular Biology(all) ,Cellular polarity ,macromolecular substances ,Biology ,Xenopus Proteins ,General Biochemistry, Genetics and Molecular Biology ,Article ,Cell biology ,Xenopus laevis ,Tubulin ,Cytoplasm ,Microtubule ,biology.protein ,Chaperone complex ,Animals ,General Agricultural and Biological Sciences ,Cytoskeleton ,Actin ,Centrioles - Abstract
Summary There are six members of the tubulin superfamily in eukaryotes [1]. Alpha- and beta-tubulin form a heterodimer that polymerizes to form microtubules, and gamma-tubulin nucleates microtubules as a component of the gamma-tubulin ring complex. Alpha-, beta-, and gamma-tubulin are conserved in all eukaryotes. In contrast, delta- and epsilon-tubulin are conserved in many, but not all, eukaryotes and are associated with centrioles, although their molecular function is unclear [2–7]. Zeta-tubulin is the sixth and final member of the tubulin superfamily and is largely uncharacterized. We find that zeta-, epsilon-, and delta-tubulin form an evolutionarily co-conserved module, the ZED module, that has been lost at several junctions in eukaryotic evolution and that zeta- and delta-tubulin are evolutionarily interchangeable. Humans lack zeta-tubulin but have delta-tubulin. In Xenopus multiciliated cells, zeta-tubulin is a component of the basal foot, a centriolar appendage that connects centrioles to the apical cytoskeleton, and co-localizes there with epsilon-tubulin. Depletion of zeta-tubulin results in disorganization of centriole distribution and polarity in multiciliated cells. In contrast with multiciliated cells, zeta-tubulin in cycling cells does not localize to centrioles and is associated with the TRiC/CCT cytoplasmic chaperone complex. We conclude that zeta-tubulin facilitates interactions between the centrioles and the apical cytoskeleton as a component of the basal foot in differentiated cells and propose that the ZED tubulins are important for centriole functionalization and orientation of centrioles with respect to cellular polarity axes. more...
- Published
- 2015
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18. The planar cell polarity effector protein Wdpcp (Fritz) controls epithelial cell cortex dynamics via septins and actomyosin
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Su Kyoung Kim, John B. Wallingford, and Tae Joo Park
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Embryo, Nonmammalian ,Biophysics ,Biology ,Septin ,Time-Lapse Imaging ,Biochemistry ,Article ,Cell membrane ,Xenopus laevis ,Imaging, Three-Dimensional ,Cell Movement ,Cell polarity ,medicine ,Animals ,Cytoskeleton ,Molecular Biology ,Glycoproteins ,Cilium ,Cell Membrane ,Intracellular Signaling Peptides and Proteins ,Cell Polarity ,Epithelial Cells ,Mesenchymal Stem Cells ,Actomyosin ,Cell Biology ,Basolateral plasma membrane ,Cell biology ,Gastrulation ,medicine.anatomical_structure ,Motile cilium ,Female ,Septins - Abstract
Planar cell polarity (PCP) signaling controls polarized behaviors in diverse tissues, including the collective cell movements of gastrulation and the planar polarized beating of motile cilia. A major question in PCP signaling concerns the mechanisms linking this signaling cascade with more general cytoskeletal elements to drive polarized behavior. Previously, we reported that the PCP effector protein Wdpcp (formerly known as Fritz) interacts with septins and is critical for collective cell migration and cilia formation. Here, we report that Wdpcp is broadly involved in maintaining cortical tension in epithelial cells. In vivo 3D time-lapse imaging revealed that Wdpcp is necessary for basolateral plasma membrane stability in epithelial tissues, and we further show that Wdpcp controls cortical septin localization to maintain cortical rigidity in mucociliary epithelial cells. Finally, we show that Wdpcp acts via actomyosin to maintain balanced cortical tension in the epithelium. These data suggest that, in addition to its role in controlling plasma membrane dynamics in collective mesenchymal cell movements, Wdpcp is also essential for normal cell cortex stability during epithelial homeostasis. more...
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- 2015
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19. Multiciliated Cells
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John B. Wallingford and Eric R. Brooks
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education.field_of_study ,Agricultural and Biological Sciences(all) ,Biochemistry, Genetics and Molecular Biology(all) ,Cilium ,Population ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Epithelium ,Cell biology ,medicine.anatomical_structure ,Deuterosome ,Microtubule ,Motile cilium ,medicine ,General Agricultural and Biological Sciences ,education ,Multiciliation - Abstract
Cilia are microtubule based cellular projections that serve a wide variety of essential functions in animal cells. Defects in cilia structure or function have recently emerged as etiological mechanisms underpinning diverse human diseases. While many eukaryotic cells possess only one or two cilia, some cells, including those of many unicellular organisms, exhibit extensive multiciliation. In vertebrates, multiciliated cells (MCCs) are a specialized population of post-mitotic cells decorated with dozens of motile cilia that beat in a polarized and synchronized fashion to drive directed fluid flow across an epithelium. Dysfunction of human MCCs is associated with diseases of the brain, airway and reproductive tracts. Despite their importance, MCCs are relatively poorly studied and we are only beginning to understand the mechanisms underlying their development and function. Here, we briefly review the general phylogeny and physiology of multiciliation and detail our current understanding of the developmental and cellular events underlying the formation, maturation, and function of MCCs in vertebrates. more...
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- 2014
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20. Systematic Discovery of Endogenous Human Ribonucleoprotein Complexes
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Kevin Drew, Edward M. Marcotte, Anna L. Mallam, Jeffrey M. Schaub, John B. Wallingford, Yu Jin Jang, Ilya J. Finkelstein, Wisath Sae-Lee, Jonghwan Kim, Anna Battenhouse, and Fan Tu
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0301 basic medicine ,Proteome ,Endogeny ,RNA-binding protein ,Computational biology ,Cell Fractionation ,Proteomics ,Interactome ,Article ,General Biochemistry, Genetics and Molecular Biology ,Mice ,03 medical and health sciences ,Replication factor C ,0302 clinical medicine ,Human disease ,Animals ,Humans ,Replication Protein C ,lcsh:QH301-705.5 ,030304 developmental biology ,Ribonucleoprotein ,0303 health sciences ,Chemistry ,Reproducibility of Results ,RNA ,Centralspindlin complex ,Embryonic stem cell ,3. Good health ,HEK293 Cells ,030104 developmental biology ,Ribonucleoproteins ,lcsh:Biology (General) ,Multiprotein Complexes ,Nucleic Acid Conformation ,Function (biology) ,030217 neurology & neurosurgery - Abstract
SUMMARY RNA-binding proteins (RBPs) play essential roles in biology and are frequently associated with human disease. Although recent studies have systematically identified individual RNA-binding proteins, their higher-order assembly into ribonucleoprotein (RNP) complexes has not been systematically investigated. Here, we describe a proteomics method for systematic identification of RNP complexes in human cells. We identify 1,428 protein complexes that associate with RNA, indicating that more than 20% of known human protein complexes contain RNA. To explore the role of RNA in the assembly of each complex, we identify complexes that dissociate, change composition, or form stable protein-only complexes in the absence of RNA. We use our method to systematically identify cell-type-specific RNA-associated proteins in mouse embryonic stem cells and finally, distribute our resource, rna.MAP, in an easy-to-use online interface (rna.proteincomplexes.org). Our system thus provides a methodology for explorations across human tissues, disease states, and throughout all domains of life., In Brief Ribonucleoprotein (RNP) complexes carry out many essential biological processes. Mallam et al. developed differential fractionation (DIF-FRAC), a proteomics method to systematically discover RNP complexes. Using their method, they discovered previously unknown RNP complexes, classified complexes by their RNA-dependent stability, and identified previously unknown roles for RNA in known protein complexes., Graphical Abstract more...
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- 2019
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21. Planar cell polarity signaling, cilia and polarized ciliary beating
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John B. Wallingford
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Cilium ,Ciliated cell ,Cell Polarity ,Context (language use) ,Cell Biology ,Biology ,Article ,Cell biology ,Planar cell polarity ,Cell polarity ,Motile cilium ,Animals ,Humans ,Cilia ,Signal transduction ,Ciliary beating ,Signal Transduction - Abstract
Planar cell polarity signaling governs a wide array of polarized cell behaviors in animals. Recent reports now show that PCP signaling is essential for the directional beating of motile cilia. Interestingly, PCP signaling acts in a variety of ciliated cell types that use motile cilia to generate directional fluid flow in very different ways. This review will synthesize these recent papers and place them in context with previous studies of PCP signaling in polarized cellular morphogenesis and collective cell movement. more...
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- 2010
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22. Identification of novel ciliogenesis factors using a new in vivo model for mucociliary epithelial development
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Atsushi Kitayama, Emily R. Herrington, Tae Joo Park, Matthew W. Grow, Naoto Ueno, Julie M. Hayes, Su Kyoung Kim, Philip B. Abitua, and John B. Wallingford
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Axoneme ,Neural Tube ,Notch ,Embryo, Nonmammalian ,Xenopus ,Xenopus Proteins ,Biology ,Models, Biological ,Epithelium ,Article ,Ciliogenesis ,medicine ,Animals ,Humans ,Basal body ,Cilia ,Molecular Biology ,Mucous Membrane ,Receptors, Notch ,Epidermis (botany) ,Gene Expression Profiling ,Cilium ,Neural tube ,TTC25 ,Gene Expression Regulation, Developmental ,Reproducibility of Results ,Mucocilliary epithelium ,Cell Biology ,biology.organism_classification ,Asthma ,Cell biology ,Mucus ,Airway ,Protein Transport ,medicine.anatomical_structure ,Epidermal Cells ,Goblet Cells ,Epidermis ,In situ hybridization ,Biomarkers ,Mig12 ,Developmental Biology - Abstract
Mucociliary epithelia are essential for homeostasis of many organs and consist of mucus-secreting goblet cells and ciliated cells. Here, we present the ciliated epidermis of Xenopus embryos as a facile model system for in vivo molecular studies of mucociliary epithelial development. Using an in situ hybridization-based approach, we identified numerous genes expressed differentially in mucus-secreting cells or in ciliated cells. Focusing on genes expressed in ciliated cells, we have identified new candidate ciliogenesis factors, including several not present in the current ciliome. We find that TTC25-GFP is localized to the base of cilia and to ciliary axonemes, and disruption of TTC25 function disrupts ciliogenesis. Mig12-GFP localizes very strongly to the base of cilia and confocal imaging of this construct allows for simple visualization of the planar polarity of basal bodies that underlies polarized ciliary beating. Knockdown of Mig12 disrupts ciliogenesis. Finally, we show that ciliogenesis factors identified in the Xenopus epidermis are required in the midline to facilitate neural tube closure. These results provide further evidence of a requirement for cilia in neural tube morphogenesis and suggest that genes identified in the Xenopus epidermis play broad roles in ciliogenesis. The suites of genes identified here will provide a foundation for future studies, and may also contribute to our understanding of pathological changes in mucociliary epithelia that accompany diseases such as asthma. more...
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- 2007
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23. Shroom Induces Apical Constriction and Is Required for Hingepoint Formation during Neural Tube Closure
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Saori L. Haigo, John B. Wallingford, Jeffrey D. Hildebrand, and Richard M. Harland
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Embryo, Nonmammalian ,Xenopus ,Nervous System ,General Biochemistry, Genetics and Molecular Biology ,medicine ,Morphogenesis ,Animals ,Actin-binding protein ,Neural Tube Defects ,Actin ,Body Patterning ,biology ,Agricultural and Biological Sciences(all) ,Convergent extension ,Biochemistry, Genetics and Molecular Biology(all) ,Microfilament Proteins ,Neural tube ,rap1 GTP-Binding Proteins ,Apical constriction ,Epithelial Cells ,Cell biology ,Neuroepithelial cell ,Actin Cytoskeleton ,Neurulation ,medicine.anatomical_structure ,Microscopy, Fluorescence ,Neural Crest ,biology.protein ,Rap1 ,General Agricultural and Biological Sciences - Abstract
Background: The morphogenetic events of early vertebrate development generally involve the combined actions of several populations of cells, each engaged in a distinct behavior. Neural tube closure, for instance, involves apicobasal cell heightening, apical constriction at hingepoints, convergent extension of the midline, and pushing by the epidermis. Although a large number of genes are known to be required for neural tube closure, in only a very few cases has the affected cell behavior been identified. For example, neural tube closure requires the actin binding protein Shroom, but the cellular basis of Shroom function and how it influences neural tube closure remain to be elucidated. Results: We show here that expression of Shroom is sufficient to organize apical constriction in transcriptionally quiescent, naive epithelial cells but not in non-polarized cells. Shroom-induced apical constriction was associated with enrichment of apically localized actin filaments and required the small GTPase Rap1 but not Rho. Endogenous Xenopus shroom was found to be expressed in cells engaged in apical constriction. Consistent with a role for Shroom in organizing apical constriction, disrupting Shroom function resulted in a specific failure of hingepoint formation, defective neuroepithelial sheet-bending, and failure of neural tube closure. Conclusions: These data demonstrate that Shroom is an essential regulator of apical constriction during neurulation. The finding that a single protein can initiate this process in epithelial cells establishes that bending of epithelial sheets may be patterned during development by the regulation of expression of single genes. more...
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- 2003
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24. Cloning and expression of Xenopus Prickle, an orthologue of a Drosophila planar cell polarity gene
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John B. Wallingford, Richard M. Harland, Ray Keller, and Toshiyasu Goto
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Embryology ,Frizzled ,animal structures ,Xenopus ,Molecular Sequence Data ,Morphogenesis ,Genes, Insect ,Pronephric duct ,Species Specificity ,Paraxial mesoderm ,Animals ,Drosophila Proteins ,Amino Acid Sequence ,Cloning, Molecular ,In Situ Hybridization ,Genetics ,Base Sequence ,Sequence Homology, Amino Acid ,biology ,Cell Polarity ,DNA ,LIM Domain Proteins ,biology.organism_classification ,Protein Structure, Tertiary ,Cell biology ,DNA-Binding Proteins ,Gastrulation ,Neurulation ,Drosophila ,Otic vesicle ,Developmental Biology - Abstract
We have cloned Xenopus orthologues of the Drosophila planar cell polarity (PCP) gene Prickle. Xenopus Prickle (XPk) is expressed in tissues at the dorsal midline during gastrulation and early neurulation. XPk is later expressed in a segmental pattern in the presomitic mesoderm and then in recently formed somites. XPk is also expressed in the tailbud, pronephric duct, retina, and the otic vesicle. The complex expression pattern of XPk suggests that PCP signaling is used in a diverse array of developmental processes in vertebrate embryos. more...
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- 2002
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25. Convergent Extension
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Richard M. Harland, Scott E. Fraser, and John B. Wallingford
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Cell signaling ,biology ,Convergent extension ,Cellular differentiation ,Embryogenesis ,Xenopus ,Morphogenesis ,Vertebrate ,Cell Biology ,Anatomy ,biology.organism_classification ,General Biochemistry, Genetics and Molecular Biology ,Cell biology ,biology.animal ,Signal transduction ,Molecular Biology ,Developmental Biology - Abstract
During development, vertebrate embryos undergo dramatic changes in shape. The lengthening and narrowing of a field of cells, termed convergent extension, contributes to a variety of morphogenetic processes. Focusing on frogs and fish, we review the different cellular mechanisms and the well-conserved signaling pathways that underlie this process. more...
- Published
- 2002
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26. Control of Intercalation Is Cell-Autonomous in the Notochord of Ciona intestinalis
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David N. Keys, John B. Wallingford, Richard M. Harland, and Michael Levine
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animal structures ,cell intercalation ,Xenopus ,Notochord ,Morphogenesis ,morphogenesis ,planar cell polarity ,Biology ,Animals, Genetically Modified ,Dishevelled ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Ciona intestinalis ,Molecular Biology ,DNA Primers ,030304 developmental biology ,Genetics ,chemistry.chemical_classification ,0303 health sciences ,Base Sequence ,Convergent extension ,fungi ,Embryo ,Cell Biology ,biology.organism_classification ,Cell biology ,Ciona ,medicine.anatomical_structure ,convergent extension ,chemistry ,Mutation ,embryonic structures ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Dishevelled signaling plays a critical role in the control of cell intercalation during convergent extension in vertebrates. This study presents evidence that Dishevelled serves a similar function in the Ciona notochord. Embryos transgenic for mutant Dishevelled fail to elongate their tails, and notochord cells fail to intercalate, though notochord cell fates are unaffected. Analysis of mosaic transgenics revealed that the effects of mutant Dishevelled on notochord intercalation are cell-autonomous in Ciona, though such defects have nonautonomous effects in Xenopus. Furthermore, our data indicate that notochord cell intercalation in Ciona does not require the progressive signals which coordinate cell intercalation in the Xenopus notochord, highlighting an important difference in how mediolateral cell intercalation is controlled in the two animals. Finally, this study establishes the Ciona embryo as an effective in vivo system for the study of the molecular control of morphogenetic cell movements in chordates. more...
- Published
- 2002
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27. Tumors in tadpoles: the Xenopus embryo as a model system for the study of tumorigenesis
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John B. Wallingford
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Xenopus ,Model system ,Xenopus Proteins ,Models, Biological ,Zinc Finger Protein GLI1 ,Animal model ,Proto-Oncogene Proteins ,Genetics ,Animals ,Humans ,Genes, Tumor Suppressor ,Oncogene Proteins ,Human T-lymphotropic virus 1 ,biology ,Nuclear Proteins ,Proto-Oncogene Proteins c-mdm2 ,Zinc Fingers ,Embryo ,Neoplasms, Experimental ,biology.organism_classification ,Cell Transformation, Neoplastic ,Carcinoma, Basal Cell ,Larva ,Trans-Activators ,Cancer research ,Tumor Suppressor Protein p53 ,Transcription Factors - Abstract
I would like to thank K. Kao for providing information prior to publication. I also thank P. Vize for comments and advice and T. Grammer, D. Hsu and M. Dionne for critical reading of the manuscript. J.B.W. is supported by NIH NRSA #1F32GM19823-01. more...
- Published
- 1999
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28. Inhibition of Morphogenetic Movement duringXenopusGastrulation by Injected Sulfatase: Implications for Anteroposterior and Dorsoventral Axis Formation
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J. Akif Uzman, Michael V. Danilchik, John B. Wallingford, and Amy K. Sater
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medicine.medical_specialty ,Time Factors ,Ultraviolet Rays ,Xenopus ,Biology ,03 medical and health sciences ,Xenopus laevis ,0302 clinical medicine ,Cell Movement ,Internal medicine ,medicine ,Morphogenesis ,Animals ,Molecular Biology ,Cells, Cultured ,030304 developmental biology ,Body Patterning ,0303 health sciences ,Dose-Response Relationship, Drug ,Convergent extension ,Sulfatase ,Embryo ,Gastrula ,Cell Biology ,Marginal zone ,biology.organism_classification ,Phenotype ,Immunohistochemistry ,Cortex (botany) ,Cell biology ,Gastrulation ,Endocrinology ,Sulfatases ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
In order to explore the role of morphogenetic movement in the establishment of anteroposterior and dorsoventral axes, we sought to identify novelin vivoinhibitors of gastrulation movements inXenopus laevis.Injection of hydrolytic sulfatase into the blastocoels of gastrula stage embryos resulted in severe anteroposterior truncation, without a corresponding truncation of the dorsoventral axis. Confocal microscopy of whole embryos revealed that gastrulation movements are severely disrupted by sulfatase; in addition, sulfatase dramatically inhibited chordomesodermal cell elongation and convergent extension movements in planar dorsal marginal zone explants. The phenotype of anteroposterior reduction elicited by sulfatase is distinctly different from commonly generated dorsoanterior phenotypes (e.g., ultraviolet irradiation of the vegetal cortex prior to cortical rotation or suramin injection), and the two varieties of phenotype appear to result from inhibition of distinct, separable components of the axis-generating machinery. more...
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- 1997
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29. Vertebrate Gastrulation: Polarity Genes Control the Matrix
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John B. Wallingford
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Polarity (international relations) ,biology ,Agricultural and Biological Sciences(all) ,Biochemistry, Genetics and Molecular Biology(all) ,Vertebrate ,Cell Polarity ,Gene Expression Regulation, Developmental ,Anatomy ,Gastrula ,Matrix (biology) ,General Biochemistry, Genetics and Molecular Biology ,Cell biology ,Extracellular Matrix ,Fibronectins ,Gastrulation ,Extracellular matrix ,Polarized cell ,biology.animal ,Vertebrates ,Animals ,Cell Surface Extensions ,General Agricultural and Biological Sciences ,Gene ,Signal Transduction - Abstract
The PCP signaling cascade controls polarized cell behaviors in various organisms. New evidence suggests that this signaling cascade also controls the deposition of extracellular matrix during vertebrate gastrulation. more...
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- 2005
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30. Fritz regulates the membrane stability mediated by septins dynamics during Convergent Extension in Xenopus embryo
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John B. Wallingford, Su Kyoung Kim, Asako Shindo, and Tae Joo Park
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Membrane ,Convergent extension ,fungi ,Dynamics (mechanics) ,Xenopus ,Embryo ,Cell Biology ,Biology ,Septin ,biology.organism_classification ,Molecular Biology ,Cell biology ,Developmental Biology - Published
- 2010
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31. S03-01. Planar cell polarity: Linking developmental regulatory mechanisms to basic cellular machinery during morphogenesis
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Su Kyoung Kim, Rick Finnell, Tae Joo Park, John B. Wallingford, Edward M. Marcotte, Srimoyee Ghosh, Philip B. Abitua, Ryan S. Gray, and Bogdan J. Wlodarczyk
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Embryology ,Planar cell polarity ,Morphogenesis ,Biology ,Cell biology ,Developmental Biology - Published
- 2009
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32. Developmental regulation of cell division mechanisms in a vertebrate embryo
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Michael Glotzer, John B. Wallingford, and Esther K. Kieserman
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Vertebrate embryo ,Cell division ,Cell Biology ,Biology ,Molecular Biology ,Cell biology ,Developmental Biology - Published
- 2008
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33. A role for Cdc42 in spindle positioning and planar orientation of cell divisions during vertebrate neural tube closure
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John B. Wallingford and Esther K. Kieserman
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biology ,Neural tube ,Closure (topology) ,Vertebrate ,Cell Biology ,CDC42 ,Orientation (graph theory) ,Cell biology ,medicine.anatomical_structure ,Planar ,biology.animal ,Spindle positioning ,medicine ,Molecular Biology ,Developmental Biology - Published
- 2009
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34. Subcellular localization and signaling properties of dishevelled in developing vertebrate embryos
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Tae Joo Park, Akira Sato, Ryan S. Gray, Raymond Habas, and John B. Wallingford
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Cytoplasm ,Frizzled ,Embryo, Nonmammalian ,Dishevelled Proteins ,Xenopus ,Xenopus Proteins ,environment and public health ,Xenopus laevis ,Cell polarity ,In Situ Hybridization ,chemistry.chemical_classification ,Agricultural and Biological Sciences(all) ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,Wnt signaling pathway ,Cell Polarity ,Vertebrate ,Embryo ,rac GTP-Binding Proteins ,Cell biology ,Dishevelled ,Protein Transport ,embryonic structures ,Intercellular Signaling Peptides and Proteins ,Female ,lipids (amino acids, peptides, and proteins) ,General Agricultural and Biological Sciences ,Plasmids ,Signal Transduction ,animal structures ,Green Fluorescent Proteins ,Cell fate determination ,General Biochemistry, Genetics and Molecular Biology ,stomatognathic system ,biology.animal ,AXIN1 ,Animals ,Molecular Biology ,Adaptor Proteins, Signal Transducing ,Biochemistry, Genetics and Molecular Biology(all) ,Convergent extension ,Cell Membrane ,fungi ,Cell Biology ,Phosphoproteins ,biology.organism_classification ,Subcellular localization ,Wnt Proteins ,chemistry ,Developmental Biology - Abstract
The Dishevelled protein mediates several diverse biological processes. Intriguingly, within the same tissues where Xenopus Dishevelled (Xdsh) controls cell fate via canonical Wnt signaling, it also controls cell polarity via the vertebrate planar cell polarity (PCP) cascade [1, 2, 3, 4, 5, 6, 7, 8 and 9]. The relationship between subcellular localization of Dishevelled and its signaling activities remains unclear; conflicting results have been reported depending upon the organism and cell types examined [8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20]. We have approached this issue by developing new reagents to sequester wild-type Dishevelled protein either at the cell membrane or away from the cell membrane. Removal of Dishevelled from the cell membrane disrupts convergent extension by preventing Rho/Rac activation and mediolateral cell polarization. By manipulating the subcellular localization of K--M (dsh1), we show that this mutation inhibits Dishevelled activation of Rac, regardless of its subcellular localization. These data demonstrate that membrane localization of Dishevelled is a prerequisite for vertebrate PCP signaling. However, both membrane-targeted and cytoplasm-targeted Dishevelled can potently activate canonical Wnt signaling, suggesting that local concentration of Dishevelled protein, but not its spatial localization, is central to canonical Wnt signaling. These results suggest that in vertebrate embryos, subcellular localization is insufficient to account for the pathway specificity of Dishevelled in the canonical Wnt versus PCP signaling cascades. more...
- Published
- 2006
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35. The Wnt signaling pathway: A focus on its role in establishing polarity of cells, tissues and organs
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John B. Wallingford and Athula H. Wikramanayake
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Focus (computing) ,Frizzled ,Polarity (physics) ,Wnt signaling pathway ,LRP6 ,LRP5 ,Cell Biology ,Biology ,Developmental Biology ,Cell biology - Published
- 2006
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36. The planar cell polarity effector Fuzzy is essential for targeted membrane trafficking, ciliogenesis, and mouse embryonic development
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Edward M. Marcotte, Ryan S. Gray, Richard H. Finnell, Phil. B. Abitua, Greg S. Weiss, Bogdan J. Wlodarczyk, John B. Wallingford, Otis Blanchard, and Insuk Lee
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Membrane ,Effector ,Ciliogenesis ,Embryogenesis ,Planar cell polarity ,Cell Biology ,Biology ,Molecular Biology ,Developmental Biology ,Cell biology - Published
- 2009
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