DOCSLIB.ORG

Acute Regulation of Tight Junction Ion Selectivity in Human Airway Epithelia

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Acute Regulation of Tight Junction Ion Selectivity in Human Airway Epithelia Acute regulation of tight junction ion selectivity in human airway epithelia Andrea N. Flynn, Omar A. Itani, Thomas O. Moninger, and Michael J. Welsh1 Howard Hughes Medical Institute, Departments of Internal Medicine and Molecular Physiology and Biophysics, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242 Contributed by Michael J. Welsh, December 31, 2008 (sent for review December 17, 2008) Electrolyte transport through and between airway epithelial cells erties of individual claudins (5, 8, 9). However, because most controls the quantity and composition of the overlying liquid. epithelia express multiple different claudins, the properties Many studies have shown acute regulation of transcellular ion conferred by individual claudins remain uncertain and appear to transport in airway epithelia. However, whether ion transport depend on context, including the cell type and presence of other through tight junctions can also be acutely regulated is poorly claudins. Moreover, claudins form homooligomers and heterooli- understood both in airway and other epithelia. To investigate the gomers, both within the same cell and across adjacent cells (8, 10). paracellular pathway, we used primary cultures of differentiated Regulation of tight junction function is also poorly under- human airway epithelia and assessed expression of claudins, the stood. Previous work on individual claudins showed that several primary determinants of paracellular permeability, and measured contain a C terminus subject to phosphorylation, which may .(transepithelial electrical properties, ion fluxes, and La3؉ move- modify claudin localization or paracellular permeability (11–14 ment. Like many other tissues, airway epithelia expressed multiple However, for the reasons outlined above, ascribing causal mech- claudins. Moreover, different cell types in the epithelium expressed anisms to specific modifications has remained difficult. Acute the same pattern of claudins. To evaluate tight junction regulation, regulation of tight junction function is even less well understood. ϩ we examined the response to histamine, an acute regulator of One example is apical Na -coupled glucose entry into small airway function. Histamine stimulated a rapid and transient in- intestinal epithelial cells, which triggered an acute increase in -crease in the paracellular Na؉ conductance, with a smaller increase paracellular permeability (15, 16). The increase involved disrup ؊ in Cl conductance. The increase was mediated by histamine H1 tion of tight junction integrity with contraction of the actomyosin .receptors and depended on an increase in intracellular Ca2؉ con- ring and dilations within the tight junctions centration. These results suggest that ion flow through the para- Previous work in airway epithelia showed that several hor- cellular pathway can be acutely regulated. Such regulation could mones, peptides, neurotransmitters, and other agents regulate facilitate coupling of the passive flow of counter ions to active electrolyte transport through the cellular pathway (1–3, 17). In transcellular transport, thereby controlling net transepithelial salt many cases, ligands alter transcellular ion transport within and water transport. seconds. Because both the cellular and paracellular pathways determine net transport, we hypothesized that the paracellular claudin ͉ histamine ͉ paracellular pathway pathway might be regulated in parallel with the cellular pathway. To test this hypothesis, we chose histamine as an agonist. First, irway epithelia determine the quantity and composition of it has a rapid and transient local effect after its release by mast Athe overlying liquid (1–3). Two parallel pathways control cells (18). Second, it can acutely regulate active transcellular ion transepithelial electrolyte transport, the cellular pathway and the transport (19, 20). Third, it is a physiological regulator of epithelial function and may be involved in airway disease (21). paracellular pathway. The cellular pathway comprises the epi- Fourth, previous studies indicated that histamine acutely in- thelial cells with their distinct apical and basolateral membranes creases transepithelial electrical conductance (G ) in airway PHYSIOLOGY containing channels, transporters, and pumps. Together, they t epithelia (19, 22). In endothelia and cell lines, activation of generate active (and sometimes passive) transepithelial ion histamine receptors disrupts the interaction of vascular endo- transport. The paracellular pathway provides a route for passive thelial-cadherin with the vimentin cytoskeleton to increase transepithelial transport, with ions moving in either direction paracellular conductance (G ) (23). Those studies focused on driven solely by their electrochemical gradient. The permeability p histamine’s effect on adherens junctions, but whether tight and ion selectivity of the paracellular pathway is critical for junction function is also altered was not addressed. Therefore, in establishing or dissipating ion concentration gradients and hence this study we tested the hypothesis that histamine might acutely for determining the ionic composition of the apical compart- regulate tight junction permeability in airway epithelia. ment and net volume flow. Thus, cellular and paracellular pathways must work in concert, being functionally matched to Results meet the transport requirements of a specific tissue. Differentiated Human Airway Epithelia Express Multiple Claudins. We The paracellular pathway is formed by tight junctions, located studied primary cultures of differentiated human airway epithe- near the apical side of the cells, and the lateral intercellular lia (24). After Ϸ2-weeks culture at the air–liquid interface, spaces, where adjacent epithelial cells interdigitate and are held epithelia contain basal cells, ciliated cells, goblet cells, and together by scattered adherens junctions (4–6). Tight junctions form the functional and structural boundary that separates apical and basolateral compartments. Importantly, they also Author contributions: A.N.F., O.A.I., T.O.M., and M.J.W. designed research; A.N.F., O.A.I., determine the ion transport properties of the paracellular path- and T.O.M. performed research; A.N.F., O.A.I., T.O.M., and M.J.W. analyzed data; and A.N.F. way. Tight junctions are comprised of a complex of proteins that and M.J.W. wrote the paper. includes claudins, which determine the ion selectivity and con- The authors declare no conflict of interest. ductance of the paracellular pathway. Freely available online through the PNAS open access option. Identification of claudin mutations associated with human 1To whom correspondence should be addressed. E-mail: [email protected]. disease have emphasized their importance in maintaining barrier This article contains supporting information online at www.pnas.org/cgi/content/full/ function and ion selectivity (7). Overexpression, knockout, 0813393106/DCSupplemental. knockdown, and mutagenesis studies have suggested the prop- © 2009 by The National Academy of Sciences of the USA www.pnas.org͞cgi͞doi͞10.1073͞pnas.0813393106 PNAS ͉ March 3, 2009 ͉ vol. 106 ͉ no. 9 ͉ 3591–3596 Downloaded by guest on October 2, 2021 +RT A -RT 40 30 Ct 20 10 0 B i ii iii Fig. 2. Claudins 1, 3, and 7 localize to tight junctions between ciliated cells (A–C) and goblet cells (D–F). Data are en face projections of Alexa Fluor 488 bound to rabbit anti-claudins 1, 3, and 7 (green) and Alexa Fluor 568 bound to antiacetylated ␣-tubulin, a ciliated cell marker (A–C; red), or to Muc5AC, a iv v vi goblet cell marker (D–F; red). Arrows point to one example of tight junctions between neighboring ciliated or goblet cells that express claudins 1, 3, or 7. Data are stacks of images, and therefore the claudin immunostaining appears slightly blurry in some panels. 60ϫ magnification. Experiments were per- formed twice with epithelia from 2 different donors. examined junctions between 2 identical cell types; if a claudin appeared at this location, we concluded that the specific cell type expressed it. We identified ciliated cells with antibodies to Fig. 1. Differentiated human airway epithelia express multiple different acetylated ␣-tubulin (27) and goblet cells with antibodies to claudins. (A) Real-time RT-PCR of primary cultures of differentiated airway epithelia. Ct indicates cycle threshold. The black and light gray bars indicate mucin Muc5AC (28). Claudins 1, 3, and 7 localized to tight samples with and without reverse transcriptase (RT), respectively. (B) Immu- junctions between neighboring ciliated cells and between neigh- nocytochemistry revealed staining of claudins 1, 3, 4, and 7 in a pattern boring goblet cells and to junctions between nonidentical cell consistent with tight junction localization (i–iv). Data are en face (Upper) and types (Fig. 2). Thus, both ciliated and goblet cells express X-Z projections (Lower) of Alexa Fluor 488 bound to rabbit anti-claudin claudins 1, 3, and 7. antibodies (green). ToPro-3 stained nuclei are in blue. Dotted line indicates position of filter. (v and vi) Immunostaining of claudin 2 was not detected in The Airway Epithelial Paracellular Pathway Is Cation-Selective. To airway epithelia (v) even though it could be detected in MDCKII cells (vi). Data investigate the paracellular pathway, we sought to eliminate the in v and vi are Alexa Fluor 488 bound to mouse anti-claudin 2 antibodies (green), and ethidium bromide-stained nuclei are in red. 40ϫ magnification. contribution of ion flow
Load more

Recommended publications
  • Vocabulario De Morfoloxía, Anatomía E Citoloxía Veterinaria
    Vocabulario de Morfoloxía, anatomía e citoloxía veterinaria (galego-español-inglés) Servizo de Normalización Lingüística Universidade de Santiago de Compostela COLECCIÓN VOCABULARIOS TEMÁTICOS N.º 4 SERVIZO DE NORMALIZACIÓN LINGÜÍSTICA Vocabulario de Morfoloxía, anatomía e citoloxía veterinaria (galego-español-inglés) 2008 UNIVERSIDADE DE SANTIAGO DE COMPOSTELA VOCABULARIO de morfoloxía, anatomía e citoloxía veterinaria : (galego-español- inglés) / coordinador Xusto A. Rodríguez Río, Servizo de Normalización Lingüística ; autores Matilde Lombardero Fernández ... [et al.]. – Santiago de Compostela : Universidade de Santiago de Compostela, Servizo de Publicacións e Intercambio Científico, 2008. – 369 p. ; 21 cm. – (Vocabularios temáticos ; 4). - D.L. C 2458-2008. – ISBN 978-84-9887-018-3 1.Medicina �������������������������������������������������������������������������veterinaria-Diccionarios�������������������������������������������������. 2.Galego (Lingua)-Glosarios, vocabularios, etc. políglotas. I.Lombardero Fernández, Matilde. II.Rodríguez Rio, Xusto A. coord. III. Universidade de Santiago de Compostela. Servizo de Normalización Lingüística, coord. IV.Universidade de Santiago de Compostela. Servizo de Publicacións e Intercambio Científico, ed. V.Serie. 591.4(038)=699=60=20 Coordinador Xusto A. Rodríguez Río (Área de Terminoloxía. Servizo de Normalización Lingüística. Universidade de Santiago de Compostela) Autoras/res Matilde Lombardero Fernández (doutora en Veterinaria e profesora do Departamento de Anatomía e Produción Animal.
    [Show full text]
  • Claudins in the Renal Collecting Duct
    International Journal of Molecular Sciences Review Claudins in the Renal Collecting Duct Janna Leiz 1,2 and Kai M. Schmidt-Ott 1,2,3,* 1 Department of Nephrology and Intensive Care Medicine, Charité-Universitätsmedizin Berlin, 12203 Berlin, Germany; [email protected] 2 Molecular and Translational Kidney Research, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), 13125 Berlin, Germany 3 Berlin Institute of Health (BIH), 10178 Berlin, Germany * Correspondence: [email protected]; Tel.: +49-(0)30-450614671 Received: 22 October 2019; Accepted: 20 December 2019; Published: 28 December 2019 Abstract: The renal collecting duct fine-tunes urinary composition, and thereby, coordinates key physiological processes, such as volume/blood pressure regulation, electrolyte-free water reabsorption, and acid-base homeostasis. The collecting duct epithelium is comprised of a tight epithelial barrier resulting in a strict separation of intraluminal urine and the interstitium. Tight junctions are key players in enforcing this barrier and in regulating paracellular transport of solutes across the epithelium. The features of tight junctions across different epithelia are strongly determined by their molecular composition. Claudins are particularly important structural components of tight junctions because they confer barrier and transport properties. In the collecting duct, a specific set of claudins (Cldn-3, Cldn-4, Cldn-7, Cldn-8) is expressed, and each of these claudins has been implicated in mediating aspects of the specific properties of its tight junction. The functional disruption of individual claudins or of the overall barrier function results in defects of blood pressure and water homeostasis. In this concise review, we provide an overview of the current knowledge on the role of the collecting duct epithelial barrier and of claudins in collecting duct function and pathophysiology.
    [Show full text]
  • Adherens Junctions, Desmosomes and Tight Junctions in Epidermal Barrier Function Johanna M
    14 The Open Dermatology Journal, 2010, 4, 14-20 Open Access Adherens Junctions, Desmosomes and Tight Junctions in Epidermal Barrier Function Johanna M. Brandner1,§, Marek Haftek*,2,§ and Carien M. Niessen3,§ 1Department of Dermatology and Venerology, University Hospital Hamburg-Eppendorf, Hamburg, Germany 2University of Lyon, EA4169 Normal and Pathological Functions of Skin Barrier, E. Herriot Hospital, Lyon, France 3Department of Dermatology, Center for Molecular Medicine, Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Germany Abstract: The skin is an indispensable barrier which protects the body from the uncontrolled loss of water and solutes as well as from chemical and physical assaults and the invasion of pathogens. In recent years several studies have suggested an important role of intercellular junctions for the barrier function of the epidermis. In this review we summarize our knowledge of the impact of adherens junctions, (corneo)-desmosomes and tight junctions on barrier function of the skin. Keywords: Cadherins, catenins, claudins, cell polarity, stratum corneum, skin diseases. INTRODUCTION ADHERENS JUNCTIONS The stratifying epidermis of the skin physically separates Adherens junctions are intercellular structures that couple the organism from its environment and serves as its first line intercellular adhesion to the cytoskeleton thereby creating a of structural and functional defense against dehydration, transcellular network that coordinate the behavior of a chemical substances, physical insults and micro-organisms. population of cells. Adherens junctions are dynamic entities The living cell layers of the epidermis are crucial in the and also function as signal platforms that regulate formation and maintenance of the barrier on two different cytoskeletal dynamics and cell polarity.
    [Show full text]
  • Cx43 and the Actin Cytoskeleton: Novel Roles and Implications for Cell-Cell Junction-Based Barrier Function Regulation
    biomolecules Review Cx43 and the Actin Cytoskeleton: Novel Roles and Implications for Cell-Cell Junction-Based Barrier Function Regulation Randy E. Strauss 1,* and Robert G. Gourdie 2,3,4,* 1 Virginia Tech, Translational Biology Medicine and Health (TBMH) Program, Roanoke, VA 24016, USA 2 Center for Heart and Reparative Medicine Research, Fralin Biomedical Research Institute at Virginia Tech Carilion, Roanoke, VA 24016, USA 3 Virginia Tech Carilion School of Medicine, Roanoke, VA 24016, USA 4 Department of Biomedical Engineering and Mechanics, Virginia Polytechnic Institute and State University, Blacksburg, VA 24060, USA * Correspondence: [email protected] (R.E.S.); [email protected] (R.G.G.) Received: 29 October 2020; Accepted: 7 December 2020; Published: 10 December 2020 Abstract: Barrier function is a vital homeostatic mechanism employed by epithelial and endothelial tissue. Diseases across a wide range of tissue types involve dynamic changes in transcellular junctional complexes and the actin cytoskeleton in the regulation of substance exchange across tissue compartments. In this review, we focus on the contribution of the gap junction protein, Cx43, to the biophysical and biochemical regulation of barrier function. First, we introduce the structure and canonical channel-dependent functions of Cx43. Second, we define barrier function and examine the key molecular structures fundamental to its regulation. Third, we survey the literature on the channel-dependent roles of connexins in barrier function, with an emphasis on the role of Cx43 and the actin cytoskeleton. Lastly, we discuss findings on the channel-independent roles of Cx43 in its associations with the actin cytoskeleton and focal adhesion structures highlighted by PI3K signaling, in the potential modulation of cellular barriers.
    [Show full text]
  • Tight Junction-Based Epithelial Microenvironment and Cell Proliferation
    Oncogene (2008) 27, 6930–6938 & 2008 Macmillan Publishers Limited All rights reserved 0950-9232/08 $32.00 www.nature.com/onc REVIEW Tight junction-based epithelial microenvironment and cell proliferation S Tsukita1, Y Yamazaki, T Katsuno, A Tamura and S Tsukita2 Laboratory of Biological Science, Graduate School of Frontier Biosciences/Graduate School of Medicine, Osaka University, Suita, Osaka, Japan Belt-like tight junctions (TJs), referred to as zonula bodies of multicellular organisms. The sheet-like divi- occludens, have long been regarded as a specialized sions allow diffusion barrier formation and selective differentiation of epithelial cell membranes. They are permeation of substances and ions (permselectivity), required for cell adhesion and paracellular barrier func- both excluding unnecessary or toxic molecules and tions, and are now thought to be partly involved in fence including necessary components to maintain home- functions and in cell polarization. Recently, the molecular ostasis. The combination of the belt-like adherens bases of TJs have gradually been unveiled. TJs are junctions (AJs) and tight junctions (TJs) have important constructed by TJ strands, whose basic frameworks are functions in the formation of epithelial cell sheets and composed of integral membrane proteins with four also in the formation of paracellular permselective transmembrane domains, designated claudins. The claudin barriers through their functions as septa (Mitic and family is supposedly composed of at least 24 members in Anderson, 1998; Tsukita and Furuse, 1999; Hartsock mice and humans. Other types of integral membrane and Nelson, 2008). Paracellular barrier functions with proteins with four transmembrane domains, namely occlu- permselectivity are unique to the epithelial cell system din and tricellulin, as well as the single transmembrane and regulate the internal homeostasis of ions and solutes proteins, JAMs (junctional adhesion molecules)and CAR in the body.
    [Show full text]
  • ILDR1 Is Important for Paracellular Water Transport and Urine Concentration Mechanism
    ILDR1 is important for paracellular water transport and urine concentration mechanism Yongfeng Gonga,1, Nina Himmerkusb,1, Abby Sunqa, Susanne Milatzb, Cosima Merkelb, Markus Bleichb,2, and Jianghui Houa,2 aDepartment of Internal Medicine, Washington University in St. Louis, St. Louis, MO 63110; and bDepartment of Physiology, University of Kiel, 24118 Kiel, Germany Edited by David W. Russell, University of Texas Southwestern Medical Center, Dallas, TX, and approved April 6, 2017 (received for review January 18, 2017) Whether the tight junction is permeable to water remains highly chromosome 12, and the sites of mutations were found to be controversial. Here, we provide evidence that the tricellular tight functionally important in making the water permeation pore (12) junction is important for paracellular water permeation and that Ig- or facilitating proper intracellular trafficking (13). Whether there like domain containing receptor 1 (ILDR1) regulates its permeability. may exist any previously unknown gene important for renal water In the mouse kidney, ILDR1 is localized to tricellular tight junctions reabsorption is a tantalizing question. Here, we have revealed Ildr1 of the distal tubules. Genetic knockout of in the mouse kidney that homozygous deletion of the ILDR1 gene, which encodes a causes polyuria and polydipsia due to renal concentrating defects. TJ protein controlling the putative paracellular water pathway, Microperfusion of live renal distal tubules reveals that they are im- can cause NDI-like phenotypes in the mouse kidney. permeable to water in normal animals but become highly perme- able to water in Ildr1 knockout animals whereas paracellular ionic Results permeabilities in the Ildr1 knockout mouse renal tubules are not The Gene Expression of ILDR1 in the Kidney.
    [Show full text]
  • Restoration of Desmosomal Junction Protein Expression and Inhibition Of
    MOLECULAR AND CLINICAL ONCOLOGY 3: 171-178, 2015 Restoration of desmosomal junction protein expression and inhibition of H3K9‑specifichistone demethylase activity by cytostatic proline‑rich polypeptide‑1 leads to suppression of tumorigenic potential in human chondrosarcoma cells KARINA GALOIAN1, AMIR QURESHI1, GINA WIDEROFF1 and H.T. TEMPLE2 1Department of Orthopaedic Surgery; 2University of Miami Tissue Bank Division, Miller School of Medicine, University of Miami, Miami, FL 33136, USA Received September 18, 2014; Accepted October 8, 2014 DOI: 10.3892/mco.2014.445 Abstract. Disruption of cell-cell junctions and the concomi- and inhibits H3K9 demethylase activity, contributing to the tant loss of polarity, downregulation of tumor-suppressive suppression of tumorigenic potential in chondrosarcoma cells. adherens junctions and desmosomes represent hallmark phenotypes for several different cancer cells. Moreover, a Introduction variety of evidence supports the argument that these two common phenotypes of cancer cells directly contribute to Chondrosarcoma is a highly aggressive bone cancer of tumorigenesis. In this study, we aimed to determine the status mesenchymal origin, which is resistant to chemo- and radia- of intercellular junction proteins expression in JJ012 human tion therapies, leaving surgical resection as the only option. malignant chondrosarcoma cells and investigate the effect Metastatic spread of malignant cells eventually develops of the antitumorigenic cytokine, proline-rich polypeptide-1 following surgical resection. The malignant progression to (PRP-1) on their expression. The cell junction pathway array chondrosarcoma has been suggested to involve a degree of data indicated downregulation of desmosomal proteins, mesenchymal-to-epithelial transition (MET) and it has been such as desmoglein (1,428-fold), desmoplakin (620-fold) and demonstrated in an in vitro model that chondrosarcoma cells plakoglobin (442-fold).
    [Show full text]
  • Regulated Assembly of Tight Junctions by Protein Kinase C ROBERT 0
    Proc. Natl. Acad. Sci. USA Vol. 92, pp. 6072-6076, June 1995 Cell Biology Regulated assembly of tight junctions by protein kinase C ROBERT 0. STUART AND SANJAY K. NIGAM* Renal Division, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115 Communicated by Carl W. Gottschalk, University of North Carolina, Chapel Hill, NC, March 14, 1995 (received for review December 7, 1994) ABSTRACT We have previously shown that protein phos- We have previously observed localized changes in intracel- phorylation plays an important role in the sorting and assembly lular calcium at sites of cell-cell contact as MDCK cells of tight junctions. We have now examined in detail the role of establish intercellular junctions and hypothesized a role for this protein kinases in intercellular junction biogenesis by using a ion in signaling relevant to intercellular-junction formation (4, combination of highly specific and broad-spectrum inhibitors 6). In addition, previous studies have suggested a role for that act by independent mechanisms. Our data indicate that phosphorylation in the assembly of tight junctions and sorting protein kinase C (PKC) is required for the proper assembly of of tight-junction proteins, zona occludens 1 (ZO-1) and cin- tight junctions. Low concentrations of the specific inhibitor of gulin, to the tight junction (3, 7). Using a combination of highly PKC, calphostin C, markedly inhibited development oftransepi- specific, as well as broad-spectrum, inhibitors of protein ki- thelial electrical resistance, a functional measure of tight- nases, we now detect apparent selective regulation of the junction biogenesis. The effect of PKC inhibitors on the devel- biogenesis oftightjunctions by protein kinase C (PKC) without opment of tight junctions, as measured by resistance, was par- immunocytochemical evidence of an effect on desmosome and alleled by a delay in the sorting ofthe tight-junction protein, zona adherens junction biogenesis.
    [Show full text]
  • Regulation of the Tight Junction Permeabilities in the TAL
    Regulation of the tight junction permeabilities in the TAL Dissertation zur Erlangung des Doktorgrades der Mathematisch-Naturwissenschaftlichen Fakultät der Christian-Albrechts-Universität zu Kiel vorgelegt von Allein Plain Reyes Kiel, 2015 “Choose a job you love, and you will never have to work a day in your life” Confucius Reviewer: Prof. Dr. Markus Bleich 2nd reviewer: Prof. Dr. Eric Beitz Committee chair: Prof. Dr. Gernot Friedrichs Committee member: Prof. Dr. Frank Melzner Oral examination: 18. 12. 2015 Approved for print: 18. 12. 2015 List of Abbreviations List of Abbreviations aTL ascending Thin Limb AVP Arginine vasopressin CaSR Calcium Sensing Receptor CD Collecting Duct Cld Claudin CLD Claudin gene CNT Connecting Tubule DCT Distal Convoluted Tubule dKO Double knock-out for Cld10 and Cld16 DMSO Dimethyl sulfoxide dTL descending Thin Limb DTT Dithiothreitol FHHNC Familial Hypomagnesemia with Hypercalciuria and Nephrocalcinosis HDAC Histone deacetylase HEPES 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid 11β-HSD2 11β-hydroxysteroid dehydrogenase type 2 Isc Equivalent short circuit current miR micro RNA MR Mineralocorticoid receptor NKCC2 Na+-K+-2Cl- cotransporter PT Proximal Tubule PTH Parathyroid hormone ROMK Renal outer medullary potassium channel Rte Transepithelial resistance SAHA Suberanilohydroxamic acid SDS Sodium dodecyl sulfate TAL Thick Ascending Limb TBS Tris-buffered saline Tween (20) Polysorbate 20 Vte Transepithelial voltage WR Water-restricted WL Water-loaded WT Wild-type iii List of Figures List of Figures Figure
    [Show full text]
  • Relocalization of Cell Adhesion Molecules During Neoplastic Transformation of Human Fibroblasts
    INTERNATIONAL JOURNAL OF ONCOLOGY 39: 1199-1204, 2011 Relocalization of cell adhesion molecules during neoplastic transformation of human fibroblasts CRISTINA BELGIOVINE, ILARIA CHIODI and CHIARA MONDELLO Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche, Via Abbiategrasso 207, 27100 Pavia, Italy Received May 6, 2011; Accepted June 10, 2011 DOI: 10.3892/ijo.2011.1119 Abstract. Studying neoplastic transformation of telomerase cell-cell contacts (1,2). Cadherins are transmembrane glyco- immortalized human fibroblasts (cen3tel), we found that the proteins mediating homotypic cell-cell adhesion via their transition from normal to tumorigenic cells was associated extracellular domain. Through their cytoplasmic domain, with the loss of growth contact inhibition, the acquisition of an they bind to catenins, which mediate the connection with the epithelial-like morphology and a change in actin organization, actin cytoskeleton. Different types of cadherins are expressed from stress fibers to cortical bundles. We show here that these in different cell types; e.g. N-cadherin is typically expressed variations were paralleled by an increase in N-cadherin expres- in mesenchymal cells, such as fibroblasts, while E-cadherin sion and relocalization of different adhesion molecules, such participates in the formation of adherens junctions in cells of as N-cadherin, α-catenin, p-120 and β-catenin. These proteins epithelial origin. The role of E-cadherin and β-catenin in the presented a clear membrane localization in tumorigenic cells development and progression of tumors of epithelial origin compared to a more diffuse, cytoplasmic distribution in is well documented (3). In particular, loosening of cell-cell primary fibroblasts and non-tumorigenic immortalized cells, contacts because of loss of E-cadherin expression and nuclear suggesting that tumorigenic cells could form strong cell-cell accumulation of β-catenin are hallmarks of the epithelial- contacts and cell contacts did not induce growth inhibition.
    [Show full text]
  • 26 April 2010 TE Prepublication Page 1 Nomina Generalia General Terms
    26 April 2010 TE PrePublication Page 1 Nomina generalia General terms E1.0.0.0.0.0.1 Modus reproductionis Reproductive mode E1.0.0.0.0.0.2 Reproductio sexualis Sexual reproduction E1.0.0.0.0.0.3 Viviparitas Viviparity E1.0.0.0.0.0.4 Heterogamia Heterogamy E1.0.0.0.0.0.5 Endogamia Endogamy E1.0.0.0.0.0.6 Sequentia reproductionis Reproductive sequence E1.0.0.0.0.0.7 Ovulatio Ovulation E1.0.0.0.0.0.8 Erectio Erection E1.0.0.0.0.0.9 Coitus Coitus; Sexual intercourse E1.0.0.0.0.0.10 Ejaculatio1 Ejaculation E1.0.0.0.0.0.11 Emissio Emission E1.0.0.0.0.0.12 Ejaculatio vera Ejaculation proper E1.0.0.0.0.0.13 Semen Semen; Ejaculate E1.0.0.0.0.0.14 Inseminatio Insemination E1.0.0.0.0.0.15 Fertilisatio Fertilization E1.0.0.0.0.0.16 Fecundatio Fecundation; Impregnation E1.0.0.0.0.0.17 Superfecundatio Superfecundation E1.0.0.0.0.0.18 Superimpregnatio Superimpregnation E1.0.0.0.0.0.19 Superfetatio Superfetation E1.0.0.0.0.0.20 Ontogenesis Ontogeny E1.0.0.0.0.0.21 Ontogenesis praenatalis Prenatal ontogeny E1.0.0.0.0.0.22 Tempus praenatale; Tempus gestationis Prenatal period; Gestation period E1.0.0.0.0.0.23 Vita praenatalis Prenatal life E1.0.0.0.0.0.24 Vita intrauterina Intra-uterine life E1.0.0.0.0.0.25 Embryogenesis2 Embryogenesis; Embryogeny E1.0.0.0.0.0.26 Fetogenesis3 Fetogenesis E1.0.0.0.0.0.27 Tempus natale Birth period E1.0.0.0.0.0.28 Ontogenesis postnatalis Postnatal ontogeny E1.0.0.0.0.0.29 Vita postnatalis Postnatal life E1.0.1.0.0.0.1 Mensurae embryonicae et fetales4 Embryonic and fetal measurements E1.0.1.0.0.0.2 Aetas a fecundatione5 Fertilization
    [Show full text]
  • Gap Junctions Between Endothelial Cells Are Disrupted by Circulating Extracellular Vesicles from Sickle Cell Patients with Acute Chest Syndrome
    International Journal of Molecular Sciences Article Gap Junctions between Endothelial Cells Are Disrupted by Circulating Extracellular Vesicles from Sickle Cell Patients with Acute Chest Syndrome Joanna Gemel , Yifan Mao, Gabrielle Lapping-Carr and Eric C. Beyer * Department of Pediatrics, University of Chicago, Chicago, IL 60637, USA; [email protected] (J.G.); [email protected] (Y.M.); [email protected] (G.L.-C.) * Correspondence: [email protected]; Tel.: +1-773-834-1498 Received: 9 October 2020; Accepted: 20 November 2020; Published: 24 November 2020 Abstract: Intercellular junctions maintain the integrity of the endothelium. We previously found that the adherens and tight junctions between endothelial cells are disrupted by plasma extracellular vesicles from patients with sickle cell disease (especially those with Acute Chest Syndrome). In the current study, we evaluated the effects of these vesicles on endothelial gap junctions. The vesicles from sickle cell patients (isolated during episodes of Acute Chest Syndrome) disrupted gap junction structures earlier and more severely than the other classes of intercellular junctions (as detected by immunofluorescence). These vesicles were much more potent than those isolated at baseline from the same subject. The treatment of endothelial cells with these vesicles led to reduced levels of connexin43 mRNA and protein. These vesicles severely reduced intercellular communication (transfer of microinjected Neurobiotin). Our data suggest a hierarchy of progressive disruption of different intercellular connections between endothelial cells by circulating extracellular vesicles that may contribute to the pathophysiology of the endothelial disturbances in sickle cell disease. Keywords: Connexin43; endothelium; gap junction; sickle cell disease; tight junction; extracellular vesicle; endothelial integrity 1.
    [Show full text]