A5059725025- Barrier Structure and Function Studies
- Connexins and lens biology
- Neurological Disease Mechanisms and Treatments
- S100 Proteins and Annexins
- Gut microbiota and health
- Caveolin-1 and cellular processes
- Cell Adhesion Molecules Research
- Clostridium difficile and Clostridium perfringens research
- Mitochondrial Function and Pathology
- Neonatal and fetal brain pathology
- Pancreatic function and diabetes
- Cellular Mechanics and Interactions
- Metabolism and Genetic Disorders
- Drug Transport and Resistance Mechanisms
- Estrogen and related hormone effects
- Salivary Gland Disorders and Functions
- Glycosylation and Glycoproteins Research
- Wnt/β-catenin signaling in development and cancer
- Intracerebral and Subarachnoid Hemorrhage Research
- Hippo pathway signaling and YAP/TAZ
- Machine Learning in Bioinformatics
- Metabolism, Diabetes, and Cancer
- Neurological diseases and metabolism
- Neuroendocrine Tumor Research Advances
- Biotin and Related Studies
National Institutes of Health
2011-2019
University of Michigan
2019
National Heart Lung and Blood Institute
2011-2018
Laboratoire de Physique du Rayonnement et de la Lumière
2013
University of North Carolina at Chapel Hill
2003-2011
University of North Carolina Health Care
2006-2008
Indiana University School of Medicine
2008
Yale University
1988-2000
University of California, Berkeley
1990
University of Pittsburgh
1983
Epithelia separate tissue spaces by regulating the passage of ions, solutes, and water through both transcellular paracellular pathways. Paracellular permeability is defined intercellular tight junctions, which vary widely among tissues with respect to solute flux, electrical resistance, ionic charge selectivity. To test hypothesis that members claudin family junction proteins create selectivity, we assessed effect reversing selected extracellular amino acids in two claudins using...
ZO-1 binds numerous transmembrane and cytoplasmic proteins is required for assembly of both adherens tight junctions, but its role in defining barrier properties an established junction unknown. We depleted MDCK cells using siRNA methods observed specific defects the large solutes, even though flux through small claudin pores was unaffected. This permeability increase accompanied by morphological alterations reorganization apical actin myosin. The defect, to a lesser extent changes, could be...
Epithelial tight junctions contain size- and charge-selective pores that control the paracellular movement of charged noncharged solutes. Claudins influence charge selectivity electrical resistance junctions, but there is no direct evidence describing pore composition or whether size density differs among cell types. To characterize independent influences from selectivity, we profiled `apparent permeabilities' (Papp) a continuous series polyethylene glycols (PEGs) across monolayers five...
Tight junctions (TJ) regulate paracellular ionic charge selectivity and conductance across epithelial tissues cell lines. These properties vary among epithelia, recent evidence implicates the claudins, a family of TJ transmembrane proteins, as important determinants both characteristics. To test hypothesis that each claudin contributes characteristic discrimination to TJ, we expressed claudins-2, -4, -11, -15 in cation-selective Madin-Darby canine kidney (MDCK) II cells anion-selective...
The structure and function of both adherens (AJ) tight (TJ) junctions are dependent on the cortical actin cytoskeleton. zonula occludens (ZO)-1 -2 proteins have context-dependent interactions with junction types bind directly to F-actin other cytoskeletal proteins, suggesting ZO-1 might regulate activity at cell junctions. To address this hypothesis, we generated stable Madin-Darby canine kidney lines depleted -2. Both paracellular permeability localization TJ disrupted in ZO-1/-2-depleted...
Tight junctions (TJs) create ion-selective paracellular permeability barriers between extracellular compartments. In the organ of Corti inner ear, TJs reticular lamina separate K + -rich endolymph and Na perilymph. humans, mutations gene encoding claudin 14 TJ protein cause profound deafness but underlying pathogenesis is unknown. To explore role in ear other tissues we created a mouse model by targeted deletion Cldn14. allele lacZ cassette expressed under Cldn14 promoter. Cldn14-lacZ...
ABSTRACT Occludin is an integral membrane protein specifically associated with tight junctions. Previous studies suggest it likely to function in forming the intercellular seal. In present study, we expressed occludin under inducible promotor occludin-null fibroblasts determine whether this confers adhesion. When human stably NRK and Rat-1 fibroblasts, which lack endogenous junctions but do have well developed ZO-1-containing adherens-like junctions, colocalizes ZO-1 points of cellcell...
Members of the large claudin family tight junction (TJ) proteins create differences in paracellular conductance and charge selectivity observed among different epithelia. Previous studies demonstrated that ionic is influenced by acidic or basic amino acids on first extracellular domain claudins. We noted two alternatively spliced variants claudin-10 database, 10a 10b, which are predicted to encode domains asked whether this might be a novel mechanism generate permselectivities from single...
The function of occludin remains elusive. Proposed roles include maintenance tight junction barriers, signaling and remodeling. To investigate a potential role in mediating cytokine-induced changes barrier properties, we measured responses to interferon-gamma plus TNFalpha control, occludin-overexpressing knockdown MDCK II monolayers. cells show complex response cytokines characterized by simultaneous increase the transepithelial electrical resistance decrease for large solutes. We observed...
Abstract Transforming growth factor‐beta 1 (TGF‐β ) is angiogenic in vivo. In two‐dimensional (2‐D) culture systems microvascular endothelial cell proliferation inhibited up to 80% by TGF‐β however, three‐dimensional (3‐D) collagen gels found have no effect on while eliciting the formation of calcium and magnesium dependent tube‐like structures mimicking angiogenesis. DNA analyses performed 3‐D cultures reveal significant difference amount or number control versus treated cultures. 2‐D known...
Palmitoylation of integral membrane proteins can affect intracellular trafficking, protein-protein interactions and protein stability. The goal the present study was to determine whether claudins, transmembrane-barrier-forming tight junction, are palmitoylated this modification has functional implications for tight-junction barrier. Claudin-14, like other members claudin family, contains membrane-proximal cysteines following both second fourth transmembrane domains, which we speculated could...
The organization and integrity of epithelial tight junctions depend on interactions between claudins, ZO scaffolding proteins, the cytoskeleton. However, although binding claudins ZO-1/2/3 numerous cytoskeletal proteins has been demonstrated in vitro, fluorescence recovery after photobleaching analysis suggests vivo are likely highly dynamic. Here we use superresolution live-cell imaging a model fibroblast system to examine relationships ZO-1, occludin, actin. We find that GFP make easily...
Biotin ligase tagging with ZO-1 was applied to identify a more complete tight junction proteome.Identical but also different proteins and functional networks were identified near the N C ends of ZO-1.The are embedded in subcompartments junction.Biotin expands proteome defines junction. The protein remain incompletely defined. Among currently known barrier-forming like occludin claudin family; scaffolding ZO-1; some cytoskeletal, signaling, cell polarity proteins. To define list infer their...
We previously identified and characterized ZO-1 as a peripheral membrane protein specifically associated with the cytoplasmic surface of tight junctions. Here we describe identification partial cDNA sequences encoding rat human their use to study assembly junctions in Caco-2 intestinal epithelial cell line. A was isolated from lambda-gtll expression library by screening mAbs. Polyclonal antibodies were raised cDNA-encoded fusion protein; several properties these support this ZO-1. Expression...
Barrett's esophagus (BE) is a specialized columnar epithelium (SCE) that develops as replacement for damaged squamous (SqE) in subjects with reflux disease, and such it apparently more acid resistant than SqE. How SCE resists injury poorly understood; one means may involve altered tight junctions (TJs) since the TJ SqE an early target of attack damage by disease. To assess this possibility, quantitative RT-PCR 21 claudins was performed on endoscopic biopsies BE from healthy without...
Addition of epidermal growth factor (EGF) to A431 human carcinoma cells results in actin reorganization and phosphorylation several cytoskeletal proteins. In the present study, we found that EGF treatment this cell line also redistribution tyrosine ZO-1. normal polarized epithelial cells, ZO-1 is restricted cytoplasmic surface most apical intercellular junctions, tight junction. contrast, majority unstimulated small subconfluent islands colocalizes with along lateral membranes rare...