DOCSLIB.ORG

Genetics of Sarcoglycan Assembly and Function 2537

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Genetics of Sarcoglycan Assembly and Function 2537 Journal of Cell Science 113, 2535-2544 (2000) 2535 Printed in Great Britain © The Company of Biologists Limited 2000 JCS1467 Differential requirement for individual sarcoglycans and dystrophin in the assembly and function of the dystrophin-glycoprotein complex Andrew A. Hack1, Man-Yee J. Lam2, Laurence Cordier3, Daria I. Shoturma3, Chantal T. Ly2, Melissa A. Hadhazy2, Michele R. Hadhazy2, H. Lee Sweeney3 and Elizabeth M. McNally2,4,* 1Department of Molecular Genetics and Cell Biology, and 2Department of Medicine, Section of Cardiology, University of Chicago, Chicago, IL, 60637, USA 3Department of Physiology, University of Pennsylvania, Philadelphia, PA, 19104, USA 4Department of Human Genetics, University of Chicago, Chicago, IL, 60637, USA *Author for correspondence (e-mail: [email protected]) Accepted 2 May; published on WWW 22 June 2000 SUMMARY Sarcoglycan is a multimeric, integral membrane sarcoglycan, α-, β- and γ-sarcoglycan were undetectable, glycoprotein complex that associates with dystrophin. while dystrophin was expressed at normal levels. In Mutations in individual sarcoglycan subunits have been contrast, without γ-sarcoglycan, reduced levels of α-, β- and identified in inherited forms of muscular dystrophy. To δ-sarcoglycan were expressed, glycosylated and formed a evaluate the contributions of sarcoglycan and dystrophin complex with each other. Thus, the elimination of γ- and δ- to muscle membrane stability and muscular dystrophy, sarcoglycan had different molecular consequences for the we compared muscle lacking specific sarcoglycans or assembly and function of the dystrophin-glycoprotein dystrophin. Here we report that mice lacking δ-sarcoglycan complex. Furthermore, these molecular differences were developed muscular dystrophy and cardiomyopathy associated with different mechanical consequences for the similar to mice lacking γ-sarcoglycan. However, unlike muscle plasma membrane. Through this in vivo analysis, a muscle lacking γ-sarcoglycan, δ-sarcoglycan-deficient model for sarcoglycan assembly is proposed. muscle was sensitive to eccentric contraction-induced disruption of the plasma membrane. In the absence of δ- Key words: Sarcoglycan, Dystrophin, Muscle, Extracellular matrix INTRODUCTION (Durbeej and Campbell, 1999; Straub et al., 1999). At the muscle plasma membrane, sarcoglycan associates with Components of the dystrophin-glycoprotein complex (DGC) dystroglycan, a second subcomplex within the DGC, and have been implicated in muscular dystrophy and dystrophin (reviewed by Hemler, 1999). Because dystrophin cardiomyopathy (reviewed by Culligan et al., 1998; Ferlini et and dystroglycan interact with actin and laminin, respectively, al., 1999; Hack et al., 2000; Ozawa et al., 1998). The absence this molecular linkage directly connects the cytoskeleton and of one or more DGC components causes progressive muscle the extracellular matrix (Ervasti and Campbell, 1993), where degeneration and leads to skeletal and cardiac muscle it is thought to provide support for the plasma membrane weakness. Specifically, mutations in human α-, β-, γ- and δ- against the forces generated during muscle contraction. sarcoglycan cause limb-girdle muscular dystrophy while Although the biochemical properties of sarcoglycan in dystrophin mutations cause Duchenne and Becker muscular normal striated muscle have been studied, less is known about dystrophy (reviewed by Bonnemann et al., 1996; Campbell, the molecular consequences of sarcoglycan deficiency for 1995). The DGC is a large multi-protein complex expressed at the assembly and function of the dystrophin-glycoprotein the plasma membrane of cardiac and skeletal myocytes. complex. Studies of patient muscle biopsies have demonstrated Sarcoglycan is an independent subcomplex within the larger that the absence of one sarcoglycan has important, but variable dystrophin-glycoprotein complex (Yoshida et al., 1994) and is consequences for the stability of the other remaining composed of at least four sarcoglycan subunits, α, β, γ and δ sarcoglycan components at the plasma membrane (reviewed by (Jung et al., 1996; Yoshida et al., 1994). γ- and δ-sarcoglycan Bushby, 1999). Likewise, mice with null mutations in specific are highly related, sharing 67% amino acid similarity (Jung et members of the sarcoglycan complex have shown variation in al., 1996; Nigro et al., 1996; Noguchi et al., 1995). The role of the pattern of sarcoglycan complex disruption depending on ε-sarcoglycan in the striated muscle sarcoglycan complex is the specific sarcoglycan mutated (Araishi et al., 1999; Coral- less clear (Ettinger et al., 1997; McNally et al., 1998). Vazquez et al., 1999; Duclos et al., 1998; Hack et al., 1998; Sarcoglycan complexes exist in multiple tissues including Liu and Engvall, 1999). Based on results obtained in a striated muscle, smooth muscle and non-muscle tissues heterologous, in vitro system it has been proposed that the 2536 A. A. Hack and others absence of any single sarcoglycan may be sufficient to of animals of carrying the δ-sarcoglycan null allele was maintained completely prevent the assembly of the sarcoglycan complex by heterozygote interbreeding. (Holt and Campbell, 1998). This is in contrast to results obtained from patient studies and may reflect the heterologous Phenotypic analysis, histology and Evans blue staining cell type used for the in vitro experiments or the specific Animals were observed frequently and weighed weekly. Survival was sarcoglycan mutants studied. Furthermore, the existence of assessed at the end of each week on a cohort of homozygous mutant (n=8) and normal mice (n=5) (129SvEms-+Ter/J, The Jackson non-tetrameric sarcoglycan complexes in tissues other than Laboratory, Bar Harbor, ME). Serum creatine kinase levels were skeletal muscle (Durbeej and Campbell, 1999; Straub et al., determined as described previously (Hack et al., 1998). Tissues for 1999) suggests that sarcoglycan subcomplexes can be stable histology were either placed in 10% neutral buffered formalin with fewer than four members. overnight and embedded in paraffin or rapidly frozen in liquid To examine the cellular and molecular consequences of nitrogen-cooled isopentane for sectioning on a cryostat. Evans blue multiple mutations in the sarcoglycan complex, we have (Sigma-Aldrich, St Louis, MO) staining of muscle in vivo was generated a targeted mutation in the murine δ-sarcoglycan performed as described previously (Hack et al., 1998). gene. The absence of δ-sarcoglycan caused skeletal and cardiac Isolated muscle mechanics muscle degeneration and resembled the dystrophic phenotype of mice lacking γ-saroclgycan (Hack et al., 1998) or dystrophin Intact extensor digitorum longus (EDL) muscles were isolated from normal (n=4) and homozygous mutant (dsg−/−) animals (n=7) at 13- (Bulfield et al., 1984). Isolated muscle mechanics revealed that δ 15 weeks of age and mechanics were performed as described -sarcoglycan-deficient muscle differed from muscle lacking previously (Hack et al., 1999). All stimulations were performed with γ-sarcoglycan in its increased sensitivity to eccentric the muscle maintained at that length at which maximum twitch force −/− contraction-induced injury. In order to further investigate the was achieved (Lo). Isolated normal and dsg EDL showed similar molecular basis for this difference in vivo we studied the mean (± s.e.m.) Lo (10.9±0.2 and 12.0±0.1, respectively) and weight assembly of the sarcoglycan complex in muscle lacking either (9.4±0.3 and 9.0±0.1, respectively). Eccentric contractions (ECC) γ-sarcoglycan, δ-sarcoglycan or dystrophin. δ-sarcoglycan- were performed in a regimen consisting of five fused tetanic deficiency resulted in the secondary elimination of α-, β-, γ- contractions (700 milliseconds) during which the muscle was and ε-sarcoglycans in all parts of the secretory pathway. In lengthened by 10% Lo over the final 200 milliseconds. A 4 minute contrast, comparative analysis of muscle lacking γ-sarcoglycan recovery period was allowed between each ECC, during which the −/− muscle was maintained at Lo. The low molecular mass dye, procion (gsg ) or dystrophin (mdx) revealed diminished, but orange (0.2% w/v; Sigma-Aldrich, St Louis, MO) was included in continued, expression of the remaining sarcoglycans. the buffer to allow for detection of sarcolemmal disruption during Moreover, these residual sarcoglycans were glycosylated and the ECC protocol. After the ECC protocol, muscles were rapidly formed a multimeric complex in the absence of dystrophin or frozen in liquid nitrogen-cooled isopentane. 10 µm frozen sections γ-sarcoglycan. These findings suggest that residual sarcoglycan were cut on a cryostat (Leica Microsystems, Germany) and mounted in gsg−/− muscle may be competent to prevent the mechanical on slides using Vectashield mounting medium (Vector Laboratories, defects seen in mdx or δ-sarcoglycan-deficient muscle. Based Burlingame, CA). Slides were examined under epifluorescence on our findings, we propose a model for the assembly, on a Zeiss Axiophot microscope (Carl Zeiss, Germany) and trafficking and function of sarcoglycan within the dystrophin- photographed. glycoprotein complex. Antibodies The mouse monoclonal antibodies to α-sarcoglycan (NCL-a-sarc), β-sarcoglycan (NCL-b-sarc) and γ-sarcoglycan (NCL-g-sarc) were MATERIALS AND METHODS purchased from Novocastra (Newcastle upon Tyne, UK). Rabbit ε − − polyclonal antibodies to dystrophin (AB6-10) and -sarcoglycan Mice lacking γ-sarcoglycan (gsg / ) or dystrophin (mdx) have been described previously (Lidov
Load more

Recommended publications
  • Γ-Sarcoglycan Deficiency Increases Cell Contractility, Apoptosis And
    Research Article 1405 γ-Sarcoglycan deficiency increases cell contractility, apoptosis and MAPK pathway activation but does not affect adhesion Maureen A. Griffin1,2, Huisheng Feng1,3, Manorama Tewari1,2, Pedro Acosta1,3, Masataka Kawana1,3, H. Lee Sweeney1,3,4 and Dennis E. Discher1,2,4,* 1Pennsylvania Muscle Institute, University of Pennsylvania Medical Center, D-700 Richards Building, 3700 Hamilton Walk, Philadelphia, PA 19104-6083, USA 2Department of Chemical and Biomolecular Engineering, Towne Building, 220 South 33rd Street, University of Pennsylvania, Philadelphia, PA 19104-6393, USA 3Department of Physiology, University of Pennsylvania Medical Center, 3700 Hamilton Walk, Philadelphia, PA 19104-6085, USA 4Graduate Group in Cell and Molecular Biology, University of Pennsylvania Medical Center, 3620 Hamilton Walk, Philadelphia, PA 19104-6058, USA *Author for correspondence (e-mail: [email protected]) Accepted 10 January 2005 Journal of Cell Science 118, 1405-1416 Published by The Company of Biologists 2005 doi:10.1242/jcs.01717 Summary The functions of γ-sarcoglycan (γSG) in normal myotubes striations were more prominent in γSG-deficient myotubes are largely unknown, however γSG is known to assemble than in normal cells. An initial phosphoscreen of more than into a key membrane complex with dystroglycan and its 12 signaling proteins revealed a number of differences deficiency is one known cause of limb-girdle muscular between normal and γSG–/– muscle, both before and after dystrophy. Previous findings of apoptosis from γSG- stretching. MAPK-pathway proteins displayed the largest deficient mice are extended here to cell culture where changes in activation, although significant phosphorylation apoptosis is seen to increase more than tenfold in γSG- also appeared for other proteins linked to hypertension.
    [Show full text]
  • Further Evidence for the Organisation of the Four Sarcoglycans Proteins Within the Dystrophin–Glycoprotein Complex
    European Journal of Human Genetics (1999) 7, 251–254 © 1999 Stockton Press All rights reserved 1018–4813/99 $12.00 t http://www.stockton-press.co.uk/ejhg SHORT REPORT Further evidence for the organisation of the four sarcoglycans proteins within the dystrophin–glycoprotein complex M Vainzof1,2, ES Moreira2, G Ferraz3, MR Passos-Bueno2, SK Marie1 and M Zatz2 1Departamento de Neurologia, FMUSP, S˜ao Paulo 2Departamento de Biologia, IB-USP, S˜ao Paulo 3Departamento de Gen´etica, UFPE, Recife, PE, Brazil Based on the pattern of distribution of the SG proteins in patients with LGMD2C and 2D, and on the observed decreased abundance of dystrophin through WB in some sarcoglycans (SG) patients, we have recently suggested that α, â and δ subunits of sarcoglycan complex might be more closely associated and that γ-SG might interact more directly with dystrophin. Two additional SG patients here reported give further support to these suggestions: an LGMD2F patient showed patchy labelling for γ-SG, despite the lack of staining of the other three SG proteins; an LGMD2C boy showed deficiency in dystrophin by means of WB and IF, comparable with an DMD manifesting carrier. These two patients represent further evidence of a closer relation of α, â and δ-SG than of γ-SG and of the possible association of γ-SG with dystrophin. In addition the LGMD2C patient illustrates the potential risk of misdiagnosis using only dystrophin analysis, in cases with no positive family history, or when DNA analysis is not informative. Keywords: sarcoglycans; muscular dystrophy;
    [Show full text]
  • Anti-SGCG / Gamma Sarcoglycan Antibody (ARG41710)
    Product datasheet [email protected] ARG41710 Package: 100 μl anti-SGCG / gamma Sarcoglycan antibody Store at: -20°C Summary Product Description Rabbit Polyclonal antibody recognizes SGCG / gamma Sarcoglycan Tested Reactivity Hu Tested Application IHC-P, IP, WB Host Rabbit Clonality Polyclonal Isotype IgG Target Name SGCG / gamma Sarcoglycan Antigen Species Human Immunogen Synthetic peptide of Human SGCG / gamma Sarcoglycan Conjugation Un-conjugated Alternate Names 35DAG; DMDA1; TYPE; SCG3; DMDA; Gamma-sarcoglycan; DAGA4; 35 kDa dystrophin-associated glycoprotein; A4; SCARMD2; Gamma-SG; LGMD2C; MAM Application Instructions Application table Application Dilution IHC-P 1:50 - 1:200 IP 1:50 WB 1:500 - 1:2000 Application Note * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist. Calculated Mw 32 kDa Observed Size ~ 32 kDa Properties Form Liquid Purification Affinity purified. Buffer PBS (pH 7.4), 150 mM NaCl, 0.02% Sodium azide and 50% Glycerol. Preservative 0.02% Sodium azide Stabilizer 50% Glycerol Storage instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use. www.arigobio.com 1/2 Note For laboratory research only, not for drug, diagnostic or other use. Bioinformation Gene Symbol SGCG Gene Full Name sarcoglycan, gamma (35kDa dystrophin-associated glycoprotein) Background This gene encodes gamma-sarcoglycan, one of several sarcolemmal transmembrane glycoproteins that interact with dystrophin.
    [Show full text]
  • Big, Bad Hearts: from Flies to Man
    COMMENTARY Big, bad hearts: From flies to man Fabrizio C. Serluca and Mark C. Fishman* Novartis Institutes for Biomedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139 enetic screens have revealed sponsible gene is unknown, so it would important molecular pathways related essential genes that guide be very useful to have a compendium of to contractility are shared and, as Wolf metazoan complexity, from candidate genes. That is what a model et al. (4) demonstrate, that mutations of fates of individual cells, to system might offer. sarcomeric proteins do lead to dimin- Gpatterning of cell arrays, to their assem- ished contractile function and cardiac bly into organs (1–3). Genetic Screens for Physiology? enlargement. Now the Ur-genetic organism Dro- One vertebrate species, the zebrafish, Of course, the physiology of the hu- sophila is making a play to provide has already been subject to screens for man heart differs in many regards genes for a next wave of biology: heart mutations, including those that from that of Drosophila. The Drosoph- integrative physiology. How do organs interfere with contractility (3). One ila heart is a tube, lacking endothe- move, beat, digest, secrete, behave, and advantage of the zebrafish embryo is lium, composed of two thin layers of interact in manners adjusted to changing its transparency, so contractility can be muscle oriented in the circumferential needs of an organism? How can such monitored visually. In that species, mu- and longitudinal directions (11). Con- features be monitored without perturb- tations in sarcomeric proteins and tractions squeeze along the tube and ing the very processes under study? novel signaling pathways have been drive perilymph in alternating direc- In a recent issue of PNAS, Wolf et al.
    [Show full text]
  • Genetic Modifiers of Hereditary Neuromuscular Disorders
    cells Article Genetic Modifiers of Hereditary Neuromuscular Disorders and Cardiomyopathy Sholeh Bazrafshan 1, Hani Kushlaf 2 , Mashhood Kakroo 1, John Quinlan 2, Richard C. Becker 1 and Sakthivel Sadayappan 1,* 1 Heart, Lung and Vascular Institute, Division of Cardiovascular Health and Disease, Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA; [email protected] (S.B.); [email protected] (M.K.); [email protected] (R.C.B.) 2 Department of Neurology and Rehabilitation Medicine, Neuromuscular Center, University of Cincinnati Gardner Neuroscience Institute, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA; [email protected] (H.K.); [email protected] (J.Q.) * Correspondence: [email protected]; Tel.: +1-513-558-7498 Abstract: Novel genetic variants exist in patients with hereditary neuromuscular disorders (NMD), including muscular dystrophy. These patients also develop cardiac manifestations. However, the association between these gene variants and cardiac abnormalities is understudied. To determine genetic modifiers and features of cardiac disease in NMD patients, we have reviewed electronic medical records of 651 patients referred to the Muscular Dystrophy Association Care Center at the University of Cincinnati and characterized the clinical phenotype of 14 patients correlating with their next-generation sequencing data. The data were retrieved from the electronic medical records of the 14 patients included in the current study and comprised neurologic and cardiac phenotype and genetic reports which included comparative genomic hybridization array and NGS. Novel associations were uncovered in the following eight patients diagnosed with Limb-girdle Muscular Dystrophy, Bethlem Myopathy, Necrotizing Myopathy, Charcot-Marie-Tooth Disease, Peripheral Citation: Bazrafshan, S.; Kushlaf, H.; Kakroo, M.; Quinlan, J.; Becker, R.C.; Polyneuropathy, and Valosin-containing Protein-related Myopathy.
    [Show full text]
  • Sarcoglycan a Mutation in Miniature Dachshund Dogs Causes Limb-Girdle Muscular Dystrophy 2D
    UC Davis UC Davis Previously Published Works Title Sarcoglycan A mutation in miniature dachshund dogs causes limb-girdle muscular dystrophy 2D. Permalink https://escholarship.org/uc/item/3r82r6wn Journal Skeletal muscle, 11(1) ISSN 2044-5040 Authors Mickelson, James R Minor, Katie M Guo, Ling T et al. Publication Date 2021-01-07 DOI 10.1186/s13395-020-00257-y Peer reviewed eScholarship.org Powered by the California Digital Library University of California Mickelson et al. Skeletal Muscle (2021) 11:2 https://doi.org/10.1186/s13395-020-00257-y RESEARCH Open Access Sarcoglycan A mutation in miniature dachshund dogs causes limb-girdle muscular dystrophy 2D James R. Mickelson1* , Katie M. Minor1, Ling T. Guo2, Steven G. Friedenberg3, Jonah N. Cullen3, Amanda Ciavarella4, Lydia E. Hambrook4, Karen M. Brenner5, Sarah E. Helmond6, Stanley L. Marks7 and G. Diane Shelton2 Abstract Background: A cohort of related miniature dachshund dogs with exercise intolerance, stiff gait, dysphagia, myoglobinuria, and markedly elevated serum creatine kinase activities were identified. Methods: Muscle biopsy histopathology, immunofluorescence microscopy, and western blotting were combined to identify the specific pathologic phenotype of the myopathy, and whole genome SNP array genotype data and whole genome sequencing were combined to determine its genetic basis. Results: Muscle biopsies were dystrophic. Sarcoglycanopathy, a form of limb-girdle muscular dystrophy, was suspected based on immunostaining and western blotting, where α, β, and γ-sarcoglycan were all absent or reduced. Genetic mapping and whole genome sequencing identified a premature stop codon mutation in the sarcoglycan A subunit gene (SGCA). Affected dachshunds were confirmed on several continents.
    [Show full text]
  • Validación Y Estudio De La Implicación De Los Genes Adrbk2, Adrb1, Adra2b, Axin 2, Atp6v1c1 Y Atp6v0e En El Carcinoma Oral De
    FACULTADE DE MEDICINA E ODONTOLOXÍA Departamento de Estomatoloxía VALIDACIÓN Y ESTUDIO DE LA IMPLICACIÓN DE LOS GENES ADRBK2, ADRB1, ADRA2B, AXIN 2, ATP6V1C1 Y ATP6V0E EN EL CARCINOMA ORAL DE CÉLULAS ESCAMOSAS MEDIANTE PCR CUANTITATIVA EN TIEMPO REAL TESIS DOCTORAL EVA MARÍA OTERO REY Santiago de Compostela, enero de 2006 FACULTAD DE MEDICINA Y ODONTOLOGÍA DEPARTAMENTO DE ESTOMATOLOGÍA VALIDACIÓN Y ESTUDIO DE LA IMPLICACIÓN DE LOS GENES ADRBK2, ADRB1, ADRA2B, AXIN 2, ATP6V1C1 Y ATP6V0E EN EL CARCINOMA ORAL DE CÉLULAS ESCAMOSAS MEDIANTE PCR CUANTITATIVA EN TIEMPO REAL Autora: EVA MARÍA OTERO REY Santiago de Compostela, 20 de Enero de 2006. D. ABEL GARCÍA GARCÍA, Profesor Titular de Cirugía Oral y Maxilofacial de la Facultad de Medicina y Odontología de la Universidad de Santiago de Compostela; D. FRANCISCO BARROS ANGUEIRA, Doctor en Biología y Jefe de Laboratorio de la Fundación Pública Galega de Medicina Xenómica del Sergas; y D. JOSÉ MANUEL SOMOZA MARTÍN, Doctor en Odontología de la Universidad de Santiago de Compostela, CERTIFICAN: Que la presente Tesis Doctoral titulada “VALIDACIÓN Y ESTUDIO DE LA IMPLICACIÓN DE LOS GENES ADRBK2, ADRB1, ADRA2B, AXIN 2, ATP6V1C1 Y ATP6V0E EN EL CARCINOMA ORAL DE CÉLULAS ESCAMOSAS MEDIANTE PCR CUANTITATIVA EN TIEMPO REAL”, ha sido elaborada por Dña. EVA MARÍA OTERO REY bajo nuestra dirección, y hallándose concluida, autorizamos su presentación a fin de que pueda ser defendida ante el Tribunal correspondiente. Y para que así conste, se expide la presente certificación en Santiago de Compostela, a 20 de Enero de 2006. Prof. Dr. D. Abel García García Dr. D. Francisco Barros Angueira Dr.
    [Show full text]
  • Pigs with δ-Sarcoglycan Deficiency Exhibit Traits of Genetic
    Laboratory Investigation (2020) 100:887–899 https://doi.org/10.1038/s41374-020-0406-7 TECHNICAL REPORT Pigs with δ-sarcoglycan deficiency exhibit traits of genetic cardiomyopathy 1,2 1 1 3 3 Hitomi Matsunari ● Michiyo Honda ● Masahito Watanabe ● Satsuki Fukushima ● Kouta Suzuki ● 3 2 1 1 2 Shigeru Miyagawa ● Kazuaki Nakano ● Kazuhiro Umeyama ● Ayuko Uchikura ● Kazutoshi Okamoto ● 1 4 3 1,2 Masaki Nagaya ● Teruhiko Toyo-oka ● Yoshiki Sawa ● Hiroshi Nagashima Received: 14 December 2019 / Revised: 19 January 2020 / Accepted: 19 January 2020 / Published online: 14 February 2020 © The Author(s) 2020. This article is published with open access Abstract Genetic cardiomyopathy is a group of intractable cardiovascular disorders involving heterogeneous genetic contribution. This heterogeneity has hindered the development of life-saving therapies for this serious disease. Genetic mutations in dystrophin and its associated glycoproteins cause cardiomuscular dysfunction. Large animal models incorporating these genetic defects are crucial for developing effective medical treatments, such as tissue regeneration and gene therapy. In the present study, we knocked out the δ-sarcoglycan (δ-SG) gene (SGCD) in domestic pig by using a combination of efficient de δ 1234567890();,: 1234567890();,: novo gene editing and somatic cell nuclear transfer. Loss of -SG expression in the SGCD knockout pigs caused a concomitant reduction in the levels of α-, β-, and γ-SG in the cardiac and skeletal sarcolemma, resulting in systolic dysfunction, myocardial tissue degeneration, and sudden death. These animals exhibited symptoms resembling human genetic cardiomyopathy and are thus promising for use in preclinical studies of next-generation therapies. Introduction dilated cardiomyopathy (DCM), arrhythmogenic right ven- tricular cardiomyopathy, left ventricular noncompaction, and Idiopathic cardiomyopathy is a serious cardiac disorder restrictive cardiomyopathy [1, 3].
    [Show full text]
  • Muscular Dystrophies Involving the Dystrophin–Glycoprotein Complex: an Overview of Current Mouse Models Madeleine Durbeej and Kevin P Campbell*
    349 Muscular dystrophies involving the dystrophin–glycoprotein complex: an overview of current mouse models Madeleine Durbeej and Kevin P Campbell* The dystrophin–glycoprotein complex (DGC) is a multisubunit dystrophies are a heterogeneous group of disorders. complex that connects the cytoskeleton of a muscle fiber to its Patients with DMD have a childhood onset phenotype and surrounding extracellular matrix. Mutations in the DGC disrupt die by their early twenties as a result of either respiratory the complex and lead to muscular dystrophy. There are a few or cardiac failure, whereas patients with BMD have naturally occurring animal models of DGC-associated moderate weakness in adulthood and may have normal life muscular dystrophy (e.g. the dystrophin-deficient mdx mouse, spans. The limb–girdle muscular dystrophies have a dystrophic golden retriever dog, HFMD cat and the highly variable onset and progression, but the unifying δ-sarcoglycan-deficient BIO 14.6 cardiomyopathic hamster) theme among the limb–girdle muscular dystrophies is the that share common genetic protein abnormalities similar to initial involvement of the shoulder and pelvic girdle those of the human disease. However, the naturally occurring muscles. Moreover, muscular dystrophies may or may not animal models only partially resemble human disease. In be associated with cardiomyopathy [1–4]. addition, no naturally occurring mouse models associated with loss of other DGC components are available. This has Combined positional cloning and candidate gene encouraged the generation of genetically engineered mouse approaches have been used to identify an increasing number models for DGC-linked muscular dystrophy. Not only have of genes that are mutated in various forms of muscular analyses of these mice led to a significant improvement in our dystrophy.
    [Show full text]
  • Dynamic Changes in the Subcellular Localization of Drosophila Β-Sarcoglycan During the Cell Cycle
    CELL STRUCTURE AND FUNCTION 31: 173–180 (2006) © 2006 by Japan Society for Cell Biology Dynamic Changes in the Subcellular Localization of Drosophila β-Sarcoglycan during the Cell Cycle Reina Hashimoto1 and Masamitsu Yamaguchi1∗ 1Department of Applied Biology and Insect Biomedical Research Center, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan ABSTRACT. One of the proposed roles of sarcoglycan is to stabilize dystrophin glycoprotein complexes in muscle sarcolemma. Involvement in signal transduction has also been proposed and abnormalities in some sarcoglycan genes are known to be responsible for muscular dystrophy. While characterization of sarcoglycans in muscle has been performed, little is known about its functions in the non-muscle tissues in which mammalian sarcoglycans are expressed. Here, we investigated temporal and spatial expression patterns of Drosophila β-sarcoglycan (dScgβ) during development by immunohistochemistry. In addition to almost ubiquitous expression in various tissues and organs, as seen for its mammalian counterpart, anti-dScgβ staining data of embryos, eye imaginal discs, and salivary glands demonstrated cytoplasmic localization during S phase in addition to plasma membrane staining. Furthermore we found that subcellular localization of dScgβ dramatically changes during mitosis through possible association with tubulin. These observations point to a complex role of sarcoglycans in non-mus- cle tissues. Key words: sarcoglycan/Drosophila/tubulin/cell cycle Introduction plex is to stabilize DGC itself by interacting with dystro- phin, dystroglycan, and other constitutive proteins (Chen et β-sarcoglycan is a member of the sarcoglycan complex. al., 2006; Crosbie et al., 1999; Durbeej et al., 2000; Straub This forms part of the dystrophin glycoprotein complex et al., 1998).
    [Show full text]
  • Factors Regulating the Function and Assembly of the Sarcoglycan Complex in Brain
    Factors regulating the function and assembly of the sarcoglycan complex in brain A thesis submitted for the Degree of Doctor of Philosophy at Cardiff University School of Medicine Francesca Carlisle 2016 Supervised by Professor Derek Blake and Professor Anthony Isles Thesis summary Myoclonus dystonia (MD) is a neurogenic movement disorder that can be caused by mutations in the SGCE gene encoding ε-sarcoglycan. ε-sarcoglycan belongs to the sarcoglycan family of cell surface-localised, single-pass transmembrane proteins originally identified in muscle where they form a heterotetrameric subcomplex of the dystrophin- associated glycoprotein complex (DGC). Mutations in the SGCA, SGCB, SGCG and SGCD genes encoding α-, β-, γ- and δ-sarcoglycan cause limb-girdle muscular dystrophy (LGMD). There is no phenotypic overlap between MD and LGMD. LGMD-associated sarcoglycan mutations impair trafficking of the entire sarcoglycan complex to the cell surface and destabilise the DGC in muscle, while MD-associated mutations typically result in loss of ε- sarcoglycan from the cell surface. This suggests cell surface ε-sarcoglycan but not other sarcoglycans is required for normal brain function. To gain insight into ε-sarcoglycan’s function(s) in the brain, immunoaffinity purification was used to identify ε-sarcoglycan- interacting proteins. Ubiquitous and brain-specific ε-sarcoglycan isoforms co-purified with three other sarcoglycans including ζ-sarcoglycan (encoded by SGCZ) from the brain. Incorporation of an LGMD-associated β-sarcoglycan mutant into the brain sarcoglycan complex impaired the formation of the βδ-sarcoglycan core but failed to abrogate the association and trafficking of ε- and ζ-sarcoglycan in heterologous cells. Both ε-sarcoglycan isoforms also co-purified with β-dystroglycan, indicating inclusion in DGC-like complexes.
    [Show full text]
  • Childhood Onset Limb-Girdle Muscular Dystrophies in the Aegean Part of Turkey
    Acta Myologica • 2018; XXXVII: p. 210-220 Childhood onset limb-girdle muscular dystrophies in the Aegean part of Turkey Uluç Yi̇ş1, Gülden Dini̇ ż 2, Filiz Hazan3, Hülya Sevcan Dai̇magüler4 5, Bahar Toklu Baysal6, Figen Baydan2, Gülçin Akinci6, Aycan Ünalp6, Gül Aktan7, Erhan Bayram1, Semra Hiz1, Cem Paketçi̇1, Derya Okur1, Erdener Özer8, Ayça Ersen Danyeli̇8, Muzaffer Polat9, Gökhan Uyanik10 11 and Sebahattin Çirak4 5 ¹ Dokuz Eylül University, School of Medicine, Department of Pediatrics, Division of Child Neurology, İzmir, Turkey; ² Neuromuscular Disease Center, Tepecik Research Hospital, İzmir, Turkey; ³ Dr Behçet Uz Children’s Research Hospital, Department of Medical Genetics, İzmir, Turkey; 4 University Hospital Cologne, Department of Pediatrics, Cologne, Germany; 5 Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany; 6 Dr Behçet Uz Children’s Research Hospital, Department of Pediatric Neurology, İzmir, Turkey; 7 Ege University, School of Medicine, Department of Pediatrics, Division of Child Neurology, İzmir, Turkey; 8 Dokuz Eylül University, School of Medicine, Department of Pathology, İzmir, Turkey; 9 Celal Bayar University, School of Medicine, Department of Pediatrics, Division of Child Neurology, Manisa, Turkey; 10 Center for Medical Genetics, Hanusch Hospital, Vienna, Austria; 11 Medical Faculty, Sigmund Freud Private University, Vienna, Austria The aim of this study is to analyze the epidemiology of the ders. They are inherited in an autosomal recessive or clinical and genetic features of childhood-onset limb-girdle dominant manner. Thirty-three recessively and domi- muscular dystrophies (LGMD) in the Aegean part of Turkey. In total fifty-six pediatric cases with LGMD followed in four nantly inherited forms have currently been identified.
    [Show full text]