DOCSLIB.ORG

Characterisation of a Mouse Model of Protocadherin 19

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Characterisation of a Mouse Model of Protocadherin 19 Characterisation of a mouse model of Protocadherin 19 A thesis submitted to Cardiff University in accordance with the requirements for the degree of Doctor of Philosophy in the discipline of Neurosciences Natalia Galindo Riera Supervisor: Isabel Martínez Garay September 2019 . Declaration This work has not been submitted in substance for any other degree or award at this or any other university or place of learning, nor is being submitted concurrently in candidature for any degree or another award. Signed.......................................................... Date ............................................... STATEMENT 1 This thesis is being submitted in partial fulfillment of the requirements for the degree of Ph.D. Signed.......................................................... Date ............................................... STATEMENT 2 This thesis is the result of my own independent work/investigation, except where otherwise stated, and the thesis has not been edited by a third party beyond what is permitted by Cardiff University’s Policy on the Use of Third Party Editors by Research Degree Students. Other sources are acknowledged by explicit references. The views expressed are my own. Signed.......................................................... Date ............................................... STATEMENT 3: I hereby give consent for my thesis, if accepted, to be available online in the University’s Open Access repository and for inter-library loan, and for the title and summary to be made available to outside organisations. Signed.......................................................... Date ........................................... I Acknowledgements Many thanks to Doctor Isabel Martínez Garay for her kind supervision and patience over the duration of my PhD. Thank you for the opportunity to work in this amazing project and helping me in taking these first steps into my career. I would also like to thank Professor Yves Barde and Doctor Stéphane for their explanations and input in the project. Special thanks to Professor Meng Li, whose enthusiastic assessment of the project was very encouraging. Also, thanks to all member of my lab for the incredible support and effort to create a positive and smooth environment. To Dr. Xinsheng Nan for the happiness and expertise provided; to Dr. Cristina Llinares Benadero and Dr. Katharina Saeuberli for their earnest help and ideas; and to Ph.D. students Spyros Merkouris, Dr. Laura Cassels, Dr. Sven Bachmann and PhD students Sylvia Newbold, Monika Sledziowska, Sarah Ateaque and Erin Wosnitzka for daily lab life help and for their energy in providing a calm and warm environment. Very special thanks to my colleagues and friends, Dr. Hayley Dingsdale, Dr. Ellen Cross, Dr. Jessica Griffiths and Dr. Shireene Kalbassi for all the help and support and for providing an environment in the office that makes you feel at home. To PhD student Yipkin for listening to my rantings and exerting a role as a driver when I stayed late at the lab. Many thanks to my summer student Niahm, for her help in the RORB and SATB2 cortical analysis. My thanks also go to the Wellcome Trust and the Welsh Government’s Sêr Cymru program, who funded this project. Last but not least, I would like to thank my parents, Carlos Galindo Pastor and Maria Riera Montero, for their invaluable and unconditional support, and my extended family and friends, for their comprehension and positive mindset during the completion of my thesis. II Several people also contributed to the completion of this thesis. I would like to thank Dr. Isabel Martinez-Garay for conducting the in utero electroporations used to study the migration in Chapter 4. Many thanks to Dr Xinsheng Nan for providing the HEK293T cell line used for the shRNA effectiveness test of Chapter 4. Special thanks to my summer student Niahm, for her contribution in the RORB and SATB2 cortical analysis of Chapter 5. Thanks also go to Dr. Jessica Griffiths, for the optimisation of the BGAL and PCDH19 antibodies used in Chapter 3. Finally, I would like to also thank Dr Stéphane Badouin for providing me with the necessary equipment to perform the open field, elevated plus maze and social interaction behavioural tests, as well as Professor Riccardo Brambilla, who kindly allowed me to use the equipment needed for the 24- hour activity test conducted in Chapter 6. III Abstract The protocadherin 19 (PCDH19) gene is encoded on the X-chromosome and its mutation causes Early Infantile Epileptic Encephalopathy 9 (EIEE9). This disorder is characterised by epilepsy in infancy or early childhood, often accompanied by cognitive impairment and behavioural disturbances of diverse severity in heterozygous females. Mosaicism of PCDH19 positive and negative cells is believed to underpin the symptoms through a mechanism of cellular interference, leading to disruption in cell-cell communication, synapse formation and hyperexcitability of neurons. However, the expression and function of the gene in mammals are still poorly understood. The aim of this thesis was to characterise a Pcdh19- knock-out (KO) mouse model, focusing on cortical migration and lamination, and on animal behaviour. An analysis of the neuronal types expressing Pcdh19 in the postnatal cortex was also undertaken. By combining RNA in situ hybridisation and immunohistochemistry, Pcdh19 was found to be expressed by a wide variety of excitatory and inhibitory neuronal types, although expression was strongest in layers II/III and Va. In utero electroporation experiments in mutant animals revealed subtle differences in the migration of Pcdh19-KO neurons but no significant changes in Pcdh19 heterozygous (HET) females. Accordingly, analysis of cortical markers by immunostaining revealed only very minor differences in the number or distribution of marker-positive cells between Pcdh19-WT and mutant animals. The results of the behavioural analysis suggested a higher sensitivity of Pcdh19-HET females to new environments and demonstrated an effect of housing on the behaviour of WT animals. Together, this study highlights the heterogeneity of Pcdh19 expressing cells and discards major roles of this protein in cortical lamination and migration. However, the behavioural changes point to alterations in circuit formation or function. These results provide an initial path to gain insight into the pathophysiology of EIEE9. IV Abbreviations µg Microgram (s) µL Microlitre (s) µm Micrometer (s) 10x Low power objective, 10x magnification 5HT3aR Ionotropic serotonin receptor 5HT3a Amp Ampicillin ANOVA Analysis of variance AP Alkaline phosphatase AS Probe Antisense probe ASD Autism spectrum disorder AUC Area under the curve BCA Bicinchoninic acid BCIP/NBT 5-bromo-4-chloro-3-indolyl-phosphate/nitro blue tetrazolium Bis-Tris Bisamino-trismethan BSA Bovine serum albumin BSNP Burst spiking non-pyramidal cells CA Cornu ammonis CB Calbindin V Cb Cerebellum CBA Chicken beta actin promoter Cdh2 Cadherin 2 cDNA Coding deoxyribonucleic acid CGE Caudal ganglionic eminence CISH Colorimetric in situ hybridization CM Conserved motifs cm2 Square centimetre (s) CMV promoter Cytomegalovirus promoter CNS Central nervous system CR Calretinin CTIP2 Chicken ovalbumin upstream promoter transcription factor interacting protein 2 CTNND1 catenin delta-1 CTX Cortex CUX1 Cut like homeobox 1 DAPI 4’,6-diamidino-2-phenylindol dihydrochloride DCX Doublecortin DG Dentate gyrus DIG Digoxigenin VI DMEM Dulbecco’s Modified Eagle Medium DMSO Dimethyl sulfoxide DNA Deoxyribonucleic acid dNTP Deoxynucleotide triphosphate dsDNA Double-stranded DNA DTT Dithiothreitol dUTP 2'-deoxyuridine 5'-triphosphate E0.5 Embryonic day 0.5 post coitum EC Extracellular cadherin domains EDTA Ethylenediaminetetraacetic acid EGFP Enhanced green fluorescent protein EIEE9 Early infantile epileptic encephalopathy 9 EPM Elevated plus maze EPS Expanded polystyrene ERα Oestrogen receptor alpha EtOH Ethanol FBS foetal bovine serum FISH Fluorescent in situ hybridization FS Fast-spiking GABA Gamma aminobutyric acid VII GAD65/67 Glutamic acid decarboxylase 65 and 67 GAPDH Glyceraldehyde 3-phosphate dehydrogenase h Hours HA-tag Human influenza hemagglutinin tag HEBS HEPES saline buffer HEK293T Human embryonic kidney cells experiment 293 transformed with large T antigen HET Heterozygous HPC Hippocampus HRP Horseradish peroxidase IHC Immunohistochemistry IP Intermediate progenitor IRES Internal ribosomal entry site IS Irregular spiking ISH In situ hybridization IUE In utero electroporation Kan Kanamycin sulphate KCl Potassium chloride kDa Kilodalton (s) KH2PO4 Potassium phosphate monobasic VIII KO Knock-out L Litre (s) LDS Lithium dodecyl sulphate LHX2 LIM homeobox 2 M Molar MES 2-(N-morpholino) ethanesulfonic acid MgCl2 Magnesium dichloride MGE Medial ganglionic eminence MGH Mixed genotype housing Min Minute (s) mL Millilitre (s) mM millimolar mM Millimolar mm Millimeter (s) MW Molecular weight MZ Marginal zone Na2HPO4 Sodium phosphate dibasic NaCl Sodium chloride NEAA Non-Essential amino acids NEC Neural stem neuroepihtelial cell IX ng Nanograms OB Olfactory bulb OCT Optimum cutting temperature OTX1 Orthodenticle homeobox 1 P0 Postnatal day 0 PBS Phosphate buffered saline Pcdh10 Protocadherin 10 Pcdh17 Protocadherin 17 Pcdh19 Rodent protocadherin19 gene PCDH19 Human protocadherin19 gene PCDH19 Protocadherin19 protein Pcdh19FL Protocadherin 19 full-length PCR Polymerase chain reaction PFA Paraformaldehyde POD Peroxidase PV Parvalbumin RGC Radial glial cell RIPA Radioimmunoprecipitation assay buffer RNA
Load more

Recommended publications
  • Δ-Protocadherin Function: from Molecular Adhesion Properties to Brain Circuitry
    δ-Protocadherin Function: From Molecular Adhesion Properties to Brain Circuitry DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Sharon Rose Cooper Graduate Program in Molecular, Cellular and Developmental Biology The Ohio State University 2017 Dissertation Committee: Dr. James Jontes, Advisor Dr. Marcos Sotomayor, Co-advisor Dr. Heithem El-Hodiri Dr. Sharon Amacher Copyrighted by Sharon Rose Cooper 2017 Abstract Selective cell-to-cell adhesion is essential for normal development of the vertebrate brain, contributing to coordinated cell movements, regional partitioning and synapse formation. Members of the cadherin superfamily mediate calcium-dependent cell adhesion, and selective adhesion by various family members is thought to contribute to the development of neural circuitry. Members of the δ-protocadherin subfamily of cadherins are differentially expressed in the vertebrate nervous system and have been implicated in a range of neurodevelopmental disorders: schizophrenia, mental retardation, and epilepsy. However, little is known about how the δ- protocadherins contribute to the development of the nervous system, nor how this development is disrupted in the disease state. Here I focus on one member of the δ-protocadherin family, protocadherin-19 (pcdh19), since it has the clearest link to a neurodevelopmental disease, being the second most clinically relevant gene in epilepsy. Using pcdh19 transgenic zebrafish, we observed columnar modules of pcdh19-expresing cells in the optic tectum. In the absence of Pcdh19, the columnar organization is disrupted and visually guided behaviors are impaired. Furthermore, similar columns were observed in pcdh1a transgenic zebrafish, located both in the tectum and in other brain regions.
    [Show full text]
  • PAPC Couples the Segmentation Clock to Somite Morphogenesis by Regulating N-Cadherin-Dependent Adhesion
    © 2017. Published by The Company of Biologists Ltd | Development (2017) 144, 664-676 doi:10.1242/dev.143974 RESEARCH ARTICLE PAPC couples the segmentation clock to somite morphogenesis by regulating N-cadherin-dependent adhesion Jérome Chal1,2,3,4,5,*, Charlenè Guillot3,4,* and Olivier Pourquié1,2,3,4,5,6,7,‡ ABSTRACT specific level of the PSM called the determination front. The Vertebrate segmentation is characterized by the periodic formation of determination front is defined as a signaling threshold epithelial somites from the mesenchymal presomitic mesoderm implemented by posterior gradients of Wnt and FGF (Aulehla (PSM). How the rhythmic signaling pulse delivered by the et al., 2003; Diez del Corral and Storey, 2004; Dubrulle et al., segmentation clock is translated into the periodic morphogenesis of 2001; Hubaud and Pourquie, 2014; Sawada et al., 2001). Cells of somites remains poorly understood. Here, we focused on the role of the posterior PSM exhibit mesenchymal characteristics and paraxial protocadherin (PAPC/Pcdh8) in this process. We showed express Snail-related transcription factors (Dale et al., 2006; that in chicken and mouse embryos, PAPC expression is tightly Nieto, 2002). In the anterior PSM, cells downregulate snail/slug regulated by the clock and wavefront system in the posterior PSM. We expression and upregulate epithelialization-promoting factors such observed that PAPC exhibits a striking complementary pattern to N- as paraxis (Barnes et al., 1997; Sosic et al., 1997). This molecular cadherin (CDH2), marking the interface of the future somite boundary transition correlates with the anterior PSM cells progressively in the anterior PSM. Gain and loss of function of PAPC in chicken acquiring epithelial characteristics (Duband et al., 1987; Martins embryos disrupted somite segmentation by altering the CDH2- et al., 2009).
    [Show full text]
  • The Immunoglobulin Superfamily Protein SYG-1 Determines the Location of Specific Synapses in C
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Cell, Vol. 112, 619–630, March 7, 2003, Copyright 2003 by Cell Press The Immunoglobulin Superfamily Protein SYG-1 Determines the Location of Specific Synapses in C. elegans Kang Shen and Cornelia I. Bargmann* man, 2001). When the motor nerve terminals reach the Department of Anatomy muscle, motor axons secrete agrin, which acts through Department of Biochemistry and Biophysics the muscle MuSK receptor to cluster acetylcholine re- Howard Hughes Medical Institute ceptors. Axon terminals also secrete neuregulin that Box 0452 stimulates the transcription of AChRs in the synaptic University of California, San Francisco nuclei and release acetylcholine that represses AChR San Francisco, California 94143 expression from extrasynaptic nuclei. Retrograde sig- nals from muscle induce the formation of a mature pre- synaptic active zone. During the development of NMJs Summary and especially in the post-injury regeneration of NMJs, a third cell type, the Schwann cell, plays an essential During nervous system development, neurons form role in defining synaptic sites. Thus, NMJ development reproducible synapses onto specific targets. Here, we involves mutual signaling between multiple cell types at analyze the development of stereotyped synapses of the synapse. the C. elegans HSNL neuron in vivo. Postsynaptic neu- The development of central nervous system synapses rons and muscles were not required for accurate syn- has been studied mostly in dissociated neuronal cul- aptic vesicle clustering in HSNL. Instead, vulval epithe- tures, where synapse formation can be initiated by con- lial cells that contact HSNL act as synaptic guidepost tact between presynaptic axons and postsynaptic neu- cells that direct HSNL presynaptic vesicles to adjacent rons.
    [Show full text]
  • Supplementary Table 1: Adhesion Genes Data Set
    Supplementary Table 1: Adhesion genes data set PROBE Entrez Gene ID Celera Gene ID Gene_Symbol Gene_Name 160832 1 hCG201364.3 A1BG alpha-1-B glycoprotein 223658 1 hCG201364.3 A1BG alpha-1-B glycoprotein 212988 102 hCG40040.3 ADAM10 ADAM metallopeptidase domain 10 133411 4185 hCG28232.2 ADAM11 ADAM metallopeptidase domain 11 110695 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 195222 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 165344 8751 hCG20021.3 ADAM15 ADAM metallopeptidase domain 15 (metargidin) 189065 6868 null ADAM17 ADAM metallopeptidase domain 17 (tumor necrosis factor, alpha, converting enzyme) 108119 8728 hCG15398.4 ADAM19 ADAM metallopeptidase domain 19 (meltrin beta) 117763 8748 hCG20675.3 ADAM20 ADAM metallopeptidase domain 20 126448 8747 hCG1785634.2 ADAM21 ADAM metallopeptidase domain 21 208981 8747 hCG1785634.2|hCG2042897 ADAM21 ADAM metallopeptidase domain 21 180903 53616 hCG17212.4 ADAM22 ADAM metallopeptidase domain 22 177272 8745 hCG1811623.1 ADAM23 ADAM metallopeptidase domain 23 102384 10863 hCG1818505.1 ADAM28 ADAM metallopeptidase domain 28 119968 11086 hCG1786734.2 ADAM29 ADAM metallopeptidase domain 29 205542 11085 hCG1997196.1 ADAM30 ADAM metallopeptidase domain 30 148417 80332 hCG39255.4 ADAM33 ADAM metallopeptidase domain 33 140492 8756 hCG1789002.2 ADAM7 ADAM metallopeptidase domain 7 122603 101 hCG1816947.1 ADAM8 ADAM metallopeptidase domain 8 183965 8754 hCG1996391 ADAM9 ADAM metallopeptidase domain 9 (meltrin gamma) 129974 27299 hCG15447.3 ADAMDEC1 ADAM-like,
    [Show full text]
  • Local Genetic and Environmental Factors in Asthma Disease Pathogenesis: Chronicity and Persistence Mechanisms
    Eur Respir J 2007; 29: 793–803 DOI: 10.1183/09031936.00087506 CopyrightßERS Journals Ltd 2007 SERIES ‘‘GENETICS OF ASTHMA AND COPD IN THE POSTGENOME ERA’’ Edited by E. von Mutius, M. Kabesch and F. Kauffmann Number 4 in this Series Local genetic and environmental factors in asthma disease pathogenesis: chronicity and persistence mechanisms S.T. Holgate*, D.E. Davies*, R.M. Powell*, P.H. Howarth*, H.M. Haitchi# and J.W. Holloway" ABSTRACT: While asthma is an inflammatory disorder of the airways usually associated with AFFILIATIONS atopy, an important additional component is involvement of the epithelium and underlying *Allergy and Inflammation Research, Division of Infection, Inflammation mesenchyme acting as a trophic unit (EMTU). and Repair, In addition to allergens, a wide range of environmental factors interact with the EMTU, such as #IIR Division and virus infections, environmental tobacco smoke and pollutants, to initiate tissue damage and "Division of Human Genetics, School aberrant repair responses that are translated into remodelling of the airways. While candidate of Medicine, Southampton General Hospital, Southampton, UK. gene association studies have revealed polymorphic variants that influence asthmatic inflamma- tion, positional cloning of previously unknown genes is identifying a high proportion of novel CORRESPONDENCE genes in the EMTU. S.T. Holgate Dipeptidyl peptidase (DPP) 10 and disintegrin and metalloproteinase (ADAM)33 are newly Allergy and Inflammation Research MP810 identified genes strongly associated with asthma that are preferentially expressed in the airway Level D epithelium and underlying mesenchyme, respectively. Centre Block Also of increasing importance is the recognition that genes associated with asthma and atopy Southampton General Hospital have important interactions with the environment through epigenetic mechanisms that influence Southampton SO16 6YD UK their expression.
    [Show full text]
  • Differential Gene Expression in Oligodendrocyte Progenitor Cells, Oligodendrocytes and Type II Astrocytes
    Tohoku J. Exp. Med., 2011,Differential 223, 161-176 Gene Expression in OPCs, Oligodendrocytes and Type II Astrocytes 161 Differential Gene Expression in Oligodendrocyte Progenitor Cells, Oligodendrocytes and Type II Astrocytes Jian-Guo Hu,1,2,* Yan-Xia Wang,3,* Jian-Sheng Zhou,2 Chang-Jie Chen,4 Feng-Chao Wang,1 Xing-Wu Li1 and He-Zuo Lü1,2 1Department of Clinical Laboratory Science, The First Affiliated Hospital of Bengbu Medical College, Bengbu, P.R. China 2Anhui Key Laboratory of Tissue Transplantation, Bengbu Medical College, Bengbu, P.R. China 3Department of Neurobiology, Shanghai Jiaotong University School of Medicine, Shanghai, P.R. China 4Department of Laboratory Medicine, Bengbu Medical College, Bengbu, P.R. China Oligodendrocyte precursor cells (OPCs) are bipotential progenitor cells that can differentiate into myelin-forming oligodendrocytes or functionally undetermined type II astrocytes. Transplantation of OPCs is an attractive therapy for demyelinating diseases. However, due to their bipotential differentiation potential, the majority of OPCs differentiate into astrocytes at transplanted sites. It is therefore important to understand the molecular mechanisms that regulate the transition from OPCs to oligodendrocytes or astrocytes. In this study, we isolated OPCs from the spinal cords of rat embryos (16 days old) and induced them to differentiate into oligodendrocytes or type II astrocytes in the absence or presence of 10% fetal bovine serum, respectively. RNAs were extracted from each cell population and hybridized to GeneChip with 28,700 rat genes. Using the criterion of fold change > 4 in the expression level, we identified 83 genes that were up-regulated and 89 genes that were down-regulated in oligodendrocytes, and 92 genes that were up-regulated and 86 that were down-regulated in type II astrocytes compared with OPCs.
    [Show full text]
  • Cell Adhesion Molecules in Normal Skin and Melanoma
    biomolecules Review Cell Adhesion Molecules in Normal Skin and Melanoma Cian D’Arcy and Christina Kiel * Systems Biology Ireland & UCD Charles Institute of Dermatology, School of Medicine, University College Dublin, D04 V1W8 Dublin, Ireland; [email protected] * Correspondence: [email protected]; Tel.: +353-1-716-6344 Abstract: Cell adhesion molecules (CAMs) of the cadherin, integrin, immunoglobulin, and selectin protein families are indispensable for the formation and maintenance of multicellular tissues, espe- cially epithelia. In the epidermis, they are involved in cell–cell contacts and in cellular interactions with the extracellular matrix (ECM), thereby contributing to the structural integrity and barrier for- mation of the skin. Bulk and single cell RNA sequencing data show that >170 CAMs are expressed in the healthy human skin, with high expression levels in melanocytes, keratinocytes, endothelial, and smooth muscle cells. Alterations in expression levels of CAMs are involved in melanoma propagation, interaction with the microenvironment, and metastasis. Recent mechanistic analyses together with protein and gene expression data provide a better picture of the role of CAMs in the context of skin physiology and melanoma. Here, we review progress in the field and discuss molecular mechanisms in light of gene expression profiles, including recent single cell RNA expression information. We highlight key adhesion molecules in melanoma, which can guide the identification of pathways and Citation: D’Arcy, C.; Kiel, C. Cell strategies for novel anti-melanoma therapies. Adhesion Molecules in Normal Skin and Melanoma. Biomolecules 2021, 11, Keywords: cadherins; GTEx consortium; Human Protein Atlas; integrins; melanocytes; single cell 1213. https://doi.org/10.3390/ RNA sequencing; selectins; tumour microenvironment biom11081213 Academic Editor: Sang-Han Lee 1.
    [Show full text]
  • Deregulation of Cell Adhesion Molecules Is Associated with Progression and Poor Outcomes in Endometrial Cancer: Analysis of the Cancer Genome Atlas Data
    1906 ONCOLOGY LETTERS 19: 1906-1914, 2020 Deregulation of cell adhesion molecules is associated with progression and poor outcomes in endometrial cancer: Analysis of The Cancer Genome Atlas data XIANGJUN HE1, SHU LEI1,2, QI ZHANG1, LIPING MA1, NA LI1 and JIANLIU WANG2 1Central Laboratory and Institute of Clinical Molecular Biology; 2Department of Obstetrics and Gynecology, Peking University People's Hospital, Beijing 100044, P.R. China Received June 18, 2019; Accepted November 15, 2019 DOI: 10.3892/ol.2020.11295 Abstract. Cell adhesion molecules (CAMs) determine the 11 times of that in those without elevation of these 6 CAMs. behavior of cancer cells during metastasis. Although some Similar results were obtained when relapse-free survival (RFS) CAMs are dysregulated in certain types of cancer and time was used during the analysis. Prognostic reliability of the are associated with cancer progression, to the best of our six-gene model was validated using data of an independent knowledge, a comprehensive study of CAMs has not been cohort from the International Cancer Genome Consortium. In undertaken, particularly in endometrial cancer (EC). In the conclusion, a combination of CAM alterations contributed to present study the expression of 225 CAMs in EC patients progression and aggressiveness of EC. The six-gene signature with various clinicopathological phenotypes were evaluated was effective for predicting worse OS and RFS in patients by statistical analysis using publicly available data from The with EC and could be complementary to the present clinical Cancer Genome Atlas database. The Kaplan-Meier method, prognostic criteria. and univariate and multivariate Cox proportional hazards regression models were used for survival analyses.
    [Show full text]
  • Protocadherin Family: Diversity, Structure, and Function Hirofumi Morishita1 and Takeshi Yagi2
    Protocadherin family: diversity, structure, and function Hirofumi Morishita1 and Takeshi Yagi2 Protocadherins are predominantly expressed in the nervous in the cadherin superfamily [3]. The protocadherin sub- system, and constitute the largest subgroup within the cadherin group has only been identified and characterized in superfamily. The recent structural elucidation of the amino- studies from the past decade. These genetic and func- terminal cadherin domain in an archetypal protocadherin tional studies have revealed a divergent cytoplasmic revealed unique and remarkable features: the lack of an domain, as well as six or seven extracellular cadherin interface for homophilic adhesiveness found in classical (EC) domains with low sequence similarities to the EC cadherins, and the presence of loop structures specific to the domains of the classical cadherin subgroup. protocadherin family. The unique features of protocadherins extend to their genomic organization. Recent findings have Knowledge of protocadherin family at the molecular level revealed unexpected allelic and combinatorial gene regulation has increased profoundly in the past few years from for clustered protocadherins, a major subgroup in the publications of the first detailed structural and functional protocadherin family. The unique structural repertoire and findings, as well as identification of unusual gene regu- unusual gene regulation of the protocadherin family may lation for this family. In the following, we shall contex- provide the molecular basis for the extraordinary diversity
    [Show full text]
  • Structural Basis of Adhesive Binding by Desmocollins and Desmogleins
    Structural basis of adhesive binding by desmocollins and desmogleins Oliver J. Harrisona,b,1, Julia Braschc,1, Gorka Lassoa,d, Phinikoula S. Katsambaa,b, Goran Ahlsena,b, Barry Honiga,b,c,d,e,f,2, and Lawrence Shapiroa,c,f,2 aDepartment of Systems Biology, Columbia University, New York, NY 10032; bHoward Hughes Medical Institute, Columbia University, New York, NY 10032; cDepartment of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032; dCenter for Computational Biology and Bioinformatics, Columbia University, New York, NY 10032; eDepartment of Medicine, Columbia University, New York, NY 10032; and fZuckerman Mind Brain Behavior Institute, Columbia University, New York, NY 10032 Contributed by Barry Honig, April 23, 2016 (sent for review January 21, 2016; reviewed by Steven C. Almo and Dimitar B. Nikolov) Desmosomes are intercellular adhesive junctions that impart dense midline, consistent with a strand-swap mode of interaction first strength to vertebrate tissues. Their dense, ordered intercellular characterized for classical cadherins (19, 20). Nevertheless, the attachments are formed by desmogleins (Dsgs) and desmocollins identity of Dscs and Dsgs in these tomographic reconstructions could (Dscs), but the nature of trans-cellular interactions between these not be determined, and atomic-resolution structures of desmosomal specialized cadherins is unclear. Here, using solution biophysics and cadherins have not been available, with the exception of an NMR coated-bead aggregation experiments, we demonstrate family-wise structure of a monomeric EC1 fragment of mouse Dsg2 with an heterophilic specificity: All Dsgs form adhesive dimers with all Dscs, artificially extended N terminus (PDB ID code 2YQG). In addition, a with affinities characteristic of each Dsg:Dsc pair.
    [Show full text]
  • Functional Role of the Overexpression of the Myelin and Lymphocyte
    Functional role of the overexpression of the myelin and lymphocyte protein MAL in Schwann cells Inauguraldissertation Zur Erlangung der Würde eines Doktors der Philosophie Vorgelegt der Philosophisch‐Naturwissenschaftlichen Fakultät der Universität Basel von Daniela Schmid aus Ramsen (SH) Basel, 2013 Genehmigt von der Philosophisch-Naturwissenschaftlichen Fakultät auf Antrag von: Prof. M.A. Rüegg (Fakultätsverantwortlicher) Prof. N. Schaeren-Wiemers (Dissertationsleiterin) Prof. J. Kapfhammer (Korreferent) Basel, den 18. Juni 2013 Prof. J. Schibler (Dekan) To Michael 1. Acknowledgments 1. ACKNOWLEDGMENTS ................................................................................................................................ 6 2. ABBREVIATIONS ......................................................................................................................................... 7 3. SUMMARY ............................................................................................................................................... 10 4. INTRODUCTION ........................................................................................................................................ 11 4.1. THE NERVOUS SYSTEM AND MYELIN SHEATH COMPOSITION ..................................................................................... 11 4.2. SCHWANN CELL ORIGIN AND LINEAGE ................................................................................................................. 12 4.3. THE FUNCTIONAL ROLE OF THE BASAL LAMINA .....................................................................................................
    [Show full text]
  • Frequent Silencing of Protocadherin 8 by Promoter Methylation, a Candidate Tumor Suppressor for Human Gastric Cancer
    ONCOLOGY REPORTS 28: 1785-1791, 2012 Frequent silencing of protocadherin 8 by promoter methylation, a candidate tumor suppressor for human gastric cancer DANJIE ZHANG, WEI ZHAO, XINHUA LIAO, TIEQIANG BI, HAIJUN LI and XIANGMING CHE Department of General Surgery, First Affiliated Hospital of Medical College of Xi'an JiaoTong University, Xi'an 710061, P.R. China Received May 10, 2012; Accepted July 30, 2012 DOI: 10.3892/or.2012.1997 Abstract. The cadherins are a family of cell surface glycopro- These abnormalities can also define biological characteristics of teins responsible for cell adhesion which play an important role gastric cancer, which can play a role in therapy (3,4). Although in cell morphology, contact inhibition and signal transduction genetic abnormalities which include gene mutation and dele- during tumorigenesis. Protocadherin 8 (PCDH8), a member tion are remarkable in causing oncogene activation and tumor of the cadherin family, has been reported to act as a tumor suppressor gene inactivation, epigenetic silence of tumor suppressor involved in oncogenesis in breast cancer. In this study, suppressor genes through aberrant promoter hypermethylation we aimed to investigate the epigenetic inactivation of PCDH8 have also been confirmed to be frequent in gastric carcinoma and its tumor suppressor function in gastric cancer. The expres- (5,6). Gene silencing by promoter hypermethylation has been sion of PCDH8 was markedly reduced or silenced in gastric affirmed in several genes in gastric cancer, including CDH1, cancer cell lines compared with normal gastric cells or tissues. which is involved in cell adhesion, and hMLH1, which is associ- Methylation of the PCDH8 gene promoter was observed in 100% ated with DNA mismatch repair and the cell cycle regulator p16.
    [Show full text]