DOCSLIB.ORG

Modelling FKRP and Fukutin Deficiency in Zebrafish

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Modelling FKRP and Fukutin Deficiency in Zebrafish Modelling FKRP and Fukutin Deficiency in Zebrafish A thesis submitted for the degree of Doctor of Philosophy at Newcastle University November 2012 Alasdair John Wood Institute of Genetic Medicine Abstract Deficiency in fukutin-related protein (FKRP) or fukutin results in aberrant glycosylation of -dystroglycan, a key receptor for basement membrane proteins. There is a broad spectrum of disorders associated with FKRP and fukutin deficiency, ranging from limb-girdle muscular dystrophy to congenital disorders such as muscle eye brain disease and Walker-Warburg syndrome (WWS). fkrp and fukutin were knocked down in the zebrafish with antisense morpholino oligonucleotides (MO). The fkrp, fukutin and dystroglycan MOs each produced a spectrum of comparable phenotypes. With each MO producing a comparable morphant phenotype on morphological examination, it was hypothesised that inferences could be made about similarities and differences in the fkrp, fukutin dystroglycan axis during zebrafish development. The morphants had abnormal muscle fibres, including disruptions of the vertical myosepta and sarcolemma. Disorganised retinal layering in the eyes was found in both fukutin and fkrp morphants. Dysplasia of the lens was observed in most fukutin morphants and some of the fkrp morphants with a severe phenotype. Structural changes in basement membranes at 1-3 days post fertilisation (dpf) were investigated. The perturbation observed across the inner limiting membranes may account for the lens dysplasia. Cell density of the granular epithelium in the photoreceptor cell layer was found to be lower in both morphants with the least density in fukutin knock-downs, which may result from a disruption of the external limiting membrane. This leads to the conclusion that fkrp and fukutin are essential for membrane integrity in the eye and muscle of developing zebrafish. A transgenic zebrafish line expressing enhanced green fluorescent protein (EGFP) in vascular endothelium from the fli-1 promoter was used to investigate early vascularisation. In all morphants, including dystroglycan knock downs, the intersegmental vessels failed to reach the dorsal longitudinal anastomotic vessel at 1dpf. Additionally, in the fkrp and fukutin morphant the eye vasculature was abnormal. Interestingly, no change was observed in the eye vasculature of the dystroglycan morphants suggesting that fkrp and fukutin may modify proteins other than α-dystroglycan in the eye. 2 Acknowledgements To Professor Volker Straub, I am very grateful for the support and encouragement you have given me during the four years of the PhD. Your acute clinical observations have given the project a fantastic depth and direction that would otherwise have been impossible to achieve. Thank you for being a great supervisor. To Dr Rita Barresi, your insight into dystroglycanopathy has throughout the PhD been a great inspiration. Thank you for always having time for discussing my work and helping me view it within the context of the wider field. To Dr Juliane Müller, the time you devoted to helping me find my feet in the lab when I first started, will always be appreciated. Thank you for always being around to answer difficult questions on zebrafish. Special thanks to, Dr Steve Laval, for always guiding me through assessments. I am very grateful for all our interesting discussions throughout the PhD. Thank you to, all my colleges in the Newcastle Muscle Team including: the lab team, clinical team, biobank team, muscle immuno analysis unit, clinical trials group and TREAT-NMD all of which have made my time in Newcastle very enjoyable. Thank you to, Lisa Hodgson for your support with confocal microscopy, always making the work enjoyable. Dr Kath White, Tracey Davey, Ben Walker and Vivian Thompson at the electron microscopy unit, it was a great place to spend the last few months of my PhD. David Burns thanks for being a great fish technician. Catherine Jepson for teaching me a host of lab techniques that would otherwise have been difficult to learn. It has been a true pleasure to work with all of you. The work was funded by the Medical Research Council as part of the MRC Centre for Neuromuscular Diseases. 3 Contents Title Page 01 Abstract 02 Acknowledgments 03 Contents 04 List of Figures 10 List of Tables 13 List of Abbreviations 14 Chapter 1 Introduction ........................................................................................... 17 1.1 Muscular Dystrophy .................................................................................... 18 1.1.1 Duchenne Muscular Dystrophy ............................................................. 18 1.1.2 Limb Girdle Muscular Dystrophy (LGMD) ............................................. 19 1.1.3 Congenital Muscular Dystrophy (CMD) ................................................ 19 1.2 Dystrophin ................................................................................................... 21 1.3 The Dystrophin Associated Glycoprotein Complex (DGC) .......................... 22 1.3.1 Dystroglycan ......................................................................................... 23 1.3.2 Glycosylation ........................................................................................ 24 1.3.3 α-Dystroglycan Structure ...................................................................... 27 1.3.4 Deglycosylation of Dystroglycan ........................................................... 28 1.3.5 The Functional Diversity of Dystroglycan.............................................. 29 1.3.6 Dystroglycan as a Bacterial and Viral Receptor .................................... 29 1.3.7 Integrins ................................................................................................ 30 1.4 Extracellular Matrix ...................................................................................... 31 1.4.1 Laminins ............................................................................................... 31 1.4.2 Basement Membranes.......................................................................... 32 1.5 Dystroglycanopathy ..................................................................................... 33 1.5.1 Primary Dystroglycanopathy ................................................................. 33 4 1.5.2 Secondary Dystroglycanopathies ......................................................... 33 1.5.3 Fukutin .................................................................................................. 34 1.5.4 Fukutin Related Protein ........................................................................ 35 1.6 DGC Mouse Models .................................................................................... 37 1.6.1 Reichert’s Membrane and the Dystroglycan Null Mouse ...................... 39 1.6.2 DG Skeletal Muscle Chimera ................................................................ 40 1.6.3 MORE DG Null Mouse.......................................................................... 41 1.6.4 Nestin-Cre/DG Null Mouse ................................................................... 42 1.6.5 FKRP Mutant Mice ............................................................................... 42 1.6.6 Fukutin Chimera Mouse........................................................................ 43 1.6.7 Fukutin Retrotransposon Mouse ........................................................... 44 1.6.8 Mouse Model Summary ........................................................................ 45 1.7 The zebrafish as a Model for MD ................................................................ 45 1.7.1 Danio rerio ............................................................................................ 45 1.7.2 Zebrafish in Muscle Research .............................................................. 46 1.7.3 Somatogenesis ..................................................................................... 47 1.7.4 Muscle Organisation ............................................................................. 51 1.7.5 Notochord Development ....................................................................... 52 1.7.6 Embryonic Origin of Zebrafish Eye ....................................................... 52 1.7.7 Adult Zebrafish Eye .............................................................................. 54 1.7.8 Vasculogenesis .................................................................................... 55 1.8 Current Approaches for Generating Stable Zebrafish Mutant Lines ............ 56 1.8.1 Zinc Finger Nucleases .......................................................................... 57 1.8.2 TALENs ................................................................................................ 61 1.8.3 Chemical Mutagenesis and the Zebrafish Mutation Project .................. 61 1.8.4 Retrovirus Strategy ............................................................................... 62 1.8.5 Tetracycline Systems............................................................................ 62 5 1.8.6 Binary Systems ..................................................................................... 63 1.9 Direction of Study ........................................................................................ 63 Chapter 2 Materials and Methods ......................................................................... 64 2.1 Animal Models ............................................................................................. 65 2.1.1 Fish strains and maintenance
Load more

Recommended publications
  • Genetic Mutations and Mechanisms in Dilated Cardiomyopathy
    Genetic mutations and mechanisms in dilated cardiomyopathy Elizabeth M. McNally, … , Jessica R. Golbus, Megan J. Puckelwartz J Clin Invest. 2013;123(1):19-26. https://doi.org/10.1172/JCI62862. Review Series Genetic mutations account for a significant percentage of cardiomyopathies, which are a leading cause of congestive heart failure. In hypertrophic cardiomyopathy (HCM), cardiac output is limited by the thickened myocardium through impaired filling and outflow. Mutations in the genes encoding the thick filament components myosin heavy chain and myosin binding protein C (MYH7 and MYBPC3) together explain 75% of inherited HCMs, leading to the observation that HCM is a disease of the sarcomere. Many mutations are “private” or rare variants, often unique to families. In contrast, dilated cardiomyopathy (DCM) is far more genetically heterogeneous, with mutations in genes encoding cytoskeletal, nucleoskeletal, mitochondrial, and calcium-handling proteins. DCM is characterized by enlarged ventricular dimensions and impaired systolic and diastolic function. Private mutations account for most DCMs, with few hotspots or recurring mutations. More than 50 single genes are linked to inherited DCM, including many genes that also link to HCM. Relatively few clinical clues guide the diagnosis of inherited DCM, but emerging evidence supports the use of genetic testing to identify those patients at risk for faster disease progression, congestive heart failure, and arrhythmia. Find the latest version: https://jci.me/62862/pdf Review series Genetic mutations and mechanisms in dilated cardiomyopathy Elizabeth M. McNally, Jessica R. Golbus, and Megan J. Puckelwartz Department of Human Genetics, University of Chicago, Chicago, Illinois, USA. Genetic mutations account for a significant percentage of cardiomyopathies, which are a leading cause of conges- tive heart failure.
    [Show full text]
  • Limb-Girdle Muscular Dystrophy
    www.ChildLab.com 800-934-6575 LIMB-GIRDLE MUSCULAR DYSTROPHY What is Limb-Girdle Muscular Dystrophy? Limb-Girdle Muscular Dystrophy (LGMD) is a group of hereditary disorders that cause progressive muscle weakness and wasting of the shoulders and pelvis (hips). There are at least 13 different genes that cause LGMD, each associated with a different subtype. Depending on the subtype of LGMD, the age of onset is variable (childhood, adolescence, or early adulthood) and can affect other muscles of the body. Many persons with LGMD eventually need the assistance of a wheelchair, and currently there is no cure. How is LGMD inherited? LGMD can be inherited by autosomal dominant (AD) or autosomal recessive (AR) modes. The AR subtypes are much more common than the AD types. Of the AR subtypes, LGMD2A (calpain-3) is the most common (30% of cases). LGMD2B (dysferlin) accounts for 20% of cases and the sarcoglycans (LGMD2C-2F) as a group comprise 25%-30% of cases. The various subtypes represent the different protein deficiencies that can cause LGMD. What testing is available for LGMD? Diagnosis of the LGMD subtypes requires biochemical and genetic testing. This information is critical, given that management of the disease is tailored to each individual and each specific subtype. Establishing the specific LGMD subtype is also important for determining inheritance and recurrence risks for the family. The first step in diagnosis for muscular dystrophy is usually a muscle biopsy. Microscopic and protein analysis of the biopsy can often predict the type of muscular dystrophy by analyzing which protein(s) is absent. A muscle biopsy will allow for targeted analysis of the appropriate LGMD gene(s) and can rule out the diagnosis of the more common dystrophinopathies (Duchenne and Becker muscular dystrophies).
    [Show full text]
  • Fukuyama-Type Congenital Muscular Dystrophy and Abnormal
    Fukuyama-type Congenital Muscular Dystrophy and Abnormal Glycosylation of α-Dystroglycan Tatsushi Toda, Kazuhiro Kobayashi, Satoshi Takeda(1), Junko Sasaki, Hiroki Kurahashi, Hiroki Kano, Masaji Tachikawa, Fan Wang, Yoshitaka Nagai, Kiyomi Taniguchi, Mariko Taniguchi, Yoshihide Sunada(2), Toshio Terashima(3), Tamao Endo(4) and Kiichiro Matsumura(5) Division of Functional Genomics, Department of Post-Genomics and Diseases, Osaka University Graduate School of Medicine, Suita, (1) Otsuka GEN Research Institute, Otsuka Pharmaceutical Co. Ltd., Tokushima, (2) Department of Neurology, Kawasaki Medical School, Kurashiki, (3) Department of Anatomy and Neurobiology, Kobe University Graduate School of Medicine, Kobe, (4) Glycobiology Research Group, Tokyo Metropolitan Institute of Gerontology, Tokyo and (5) Department of Neurology, Teikyo University School of Medicine, Tokyo, Japan Abstract Fukuyama-type congenital muscular dystrophy (FCMD), Walker-Warburg syndrome (WWS), and muscle-eye-brain (MEB) disease are clinically similar autosomal recessive disorders characterized by congenital muscular dystrophy, lissencephaly, and eye anomalies. We identified the gene for FCMD and MEB, which encodes the fukutin protein and the protein O-linked mannose β1, 2-N-acetylglucosaminyltransferase (POMGnT1), respectively. α-dystroglycan is a key component of the dystrophin-glycoprotein-complex, providing a tight linkage between the cell and basement membranes by binding laminin via its carbohydrate residues. Recent studies have revealed that posttranslational
    [Show full text]
  • Muscle Diseases: the Muscular Dystrophies
    ANRV295-PM02-04 ARI 13 December 2006 2:57 Muscle Diseases: The Muscular Dystrophies Elizabeth M. McNally and Peter Pytel Department of Medicine, Section of Cardiology, University of Chicago, Chicago, Illinois 60637; email: [email protected] Department of Pathology, University of Chicago, Chicago, Illinois 60637; email: [email protected] Annu. Rev. Pathol. Mech. Dis. 2007. Key Words 2:87–109 myotonia, sarcopenia, muscle regeneration, dystrophin, lamin A/C, The Annual Review of Pathology: Mechanisms of Disease is online at nucleotide repeat expansion pathmechdis.annualreviews.org Abstract by Drexel University on 01/13/13. For personal use only. This article’s doi: 10.1146/annurev.pathol.2.010506.091936 Dystrophic muscle disease can occur at any age. Early- or childhood- onset muscular dystrophies may be associated with profound loss Copyright c 2007 by Annual Reviews. All rights reserved of muscle function, affecting ambulation, posture, and cardiac and respiratory function. Late-onset muscular dystrophies or myopathies 1553-4006/07/0228-0087$20.00 Annu. Rev. Pathol. Mech. Dis. 2007.2:87-109. Downloaded from www.annualreviews.org may be mild and associated with slight weakness and an inability to increase muscle mass. The phenotype of muscular dystrophy is an endpoint that arises from a diverse set of genetic pathways. Genes associated with muscular dystrophies encode proteins of the plasma membrane and extracellular matrix, and the sarcomere and Z band, as well as nuclear membrane components. Because muscle has such distinctive structural and regenerative properties, many of the genes implicated in these disorders target pathways unique to muscle or more highly expressed in muscle.
    [Show full text]
  • Table E-2. Distinguishing Features of Muscular Dystrophies Designation
    Table e-2. Distinguishing features of muscular dystrophies Designation Protein Chromosome Inheritance Common Distinguishing Distinguishing EMG Complications patterns of clinical features and muscle biopsy weakness features X-linked muscular dystrophies EDMD-X1 Emerin Xq28 XR HP Contractures Cardiac invariably present conduction early in the disease abnormalities course EDMD-X2 FHL1 Xq27.2 XR LG, HP or Foot drop; extremity Myofibrillar myopathy Severe SP, DM contractures and or reducing bodies respiratory neck contractures failure in many common patients Becker Dystrophin Xp21 XR LG Calf hypertrophy Mosaic appearance of Dilated muscular dystrophin on cardiomyopathy dystrophy immunohistochemistry in 4% to 70% in affected woman depending on disease duration Limb-girdle muscular dystrophies (LGMDs) LGMD1A Myotilin 5q22.3-31.3 AD LG, DM Onset > 40 years, Myofibrillar myopathy, Cardiomyopathy, foot drop, myotonic or respiratory asymmetric muscle pseudomyotonic muscle weakness weakness and discharges atrophy LGMD1B Lamin A/C 1q11-21 AD LG, HP, DM Early Cardiomyopathy humeroperoneal and conduction weakness, limb system disease contractures LGMD1C Caveolin-3 3p25 AD LG Rippling muscles, percussion-induced rapid contractions, prominent muscle cramps and calf hypertrophy LGMD1D DNAJB6 6q23 AD LG, DM Onset > 40 years, Myofibrillar myopathy, foot drop rimmed vacuoles, myotonic or pseudomyotonic discharges LGMD1E Desmin 2q35 AD LG, HP, DM Onset < 40 years, Myofibrillar myopathy, Cardiomyopathy, foot drop myotonic or respiratory pseudomyotonic muscle
    [Show full text]
  • The Congenital and Limb-Girdle Muscular Dystrophies Sharpening the Focus, Blurring the Boundaries
    NEUROLOGICAL REVIEW SECTION EDITOR: DAVID E. PLEASURE, MD The Congenital and Limb-Girdle Muscular Dystrophies Sharpening the Focus, Blurring the Boundaries Janbernd Kirschner, MD; Carsten G. Bo¨nnemann, MD uring the past decade, outstanding progress in the areas of congenital and limb- girdle muscular dystrophies has led to staggering clinical and genetic complexity. With the identification of an increasing number of genetic defects, individual enti- ties have come into sharper focus and new pathogenic mechanisms for muscular dys- Dtrophies, like defects of posttranslational O-linked glycosylation, have been discovered. At the same time, this progress blurs the traditional boundaries between the categories of congenital and limb- girdle muscular dystrophies, as well as between limb-girdle muscular dystrophies and other clini- cal entities, as mutations in genes such as fukutin-related protein, dysferlin, caveolin-3 and lamin A/C can cause a striking variety of phenotypes. We reviewed the different groups of proteins cur- rently recognized as being involved in congenital and limb-girdle muscular dystrophies, associ- ated them with the clinical phenotypes, and determined some clinical and molecular clues that are helpful in the diagnostic approach to these patients. Arch Neurol. 2004;61:189-199 Muscular dystrophies were first recog- phy. The age at onset may range from early nized as a disease entity with the detailed childhood to late adulthood.5 description of the clinical presentation of During the past decade, exciting Duchenne muscular dystrophy in 1852 and progress has been made in the field of CMD thereafter.1,2 About 50 years later, Batten3 and LGMD, emphasizing differences as published the first case reports of a con- well as commonalities between them.
    [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]
  • A Homozygous Nonsense Mutation in the Fukutin Gene Causes a Walker
    845 LETTER TO JMG J Med Genet: first published as 10.1136/jmg.40.11.845 on 19 November 2003. Downloaded from A homozygous nonsense mutation in the Fukutin gene causes a Walker-Warburg syndrome phenotype D Beltra´n-Valero de Bernabe´, H van Bokhoven, E van Beusekom, W Van den Akker, S Kant, W B Dobyns, B Cormand, S Currier, B Hamel, B Talim, H Topaloglu, H G Brunner ............................................................................................................................... J Med Genet 2003;40:845–848 euronal migration is a key process in the development basis of the severity of the phenotype and the presence of of the cerebral cortex. During neocortex lamination syndrome specific symptoms (table 1). WWS is the most Nnew sets of neurones proliferate at the subventricular severe syndrome of the group, especially with regard to the zone and migrate alongside specialised radial glial fibres to brain phenotype. The WWS brain manifests cobblestone occupy their final destinations in an ‘‘inside-out’’ fashion.1 lissencephaly with agenesis of the corpus callosum, fusion of More than 25 neuronal migration disorders resulting in death hemispheres, hydrocephalus, dilatation of the fourth ven- or improper positioning of the cortical neurones have been tricle, cerebellar hypoplasia, hydrocephalus, and sometimes described in humans.2 In the cobblestone neocortex the encephalocele.34 postmitotic neurones do not respond to their stop signals, Causative genes for WWS (POMT1)5, FCMD (Fukutin)6 and and, crossing through the neocortex, bypass the glia limitans MEB (POMGnT1)7 have been identified. WWS is genetically and invade the subarachnoid space. The resulting cortex is heterogeneous5, and approximately 20% of the patients show chaotically structured, consisting of an irregular lissencepha- POMT1 mutations.
    [Show full text]
  • Mitochondrial Dysfunction in Skeletal Muscle of Fukutin Deficient Mice Is Resistant to Exercise- and AICAR-Induced Rescue
    bioRxiv preprint doi: https://doi.org/10.1101/2020.05.27.118844; this version posted May 30, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Mitochondrial dysfunction in skeletal muscle of fukutin deficient mice is resistant to exercise- and AICAR-induced rescue W. Michael Southern1,2, Anna S. Nichenko1,2, Anita E. Qualls, Kensey Portman3, Ariel Gidon3, Aaron Beedle3,4, Jarrod A. Call1,2* 1 Department of Kinesiology, University of Georgia, Athens, GA 30602, USA 2 Regenerative Bioscience Center, University of Georgia, Athens, GA 30602, USA 3. Department of Pharmaceutical Sciences, SUNY at Binghamton, Binghamton NY 13902, USA 4. Department of Pharmaceutical and Biomedical Sciences, University of Georgia, Athens, GA 30602 USA. Running title: Fukutin and mitochondrial dysfunction *Author to whom correspondence should be addressed: Jarrod A. Call, PhD, 330 River Road, Ramsey Center, Department of Kinesiology, University of Georgia, Athens, GA 30602, USA; E-mail: [email protected], Tel: +1-706-542-0636, Fax: +1-706- 542-3148 Word count: Number of figures bioRxiv preprint doi: https://doi.org/10.1101/2020.05.27.118844; this version posted May 30, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Abstract Disruptions in the dystrophin-glycoprotein complex (DGC) are clearly the primary basis underlying various forms of muscular dystrophies and dystroglycanopathies, but the cellular consequences of DGC disruption are still being investigated.
    [Show full text]
  • The Role of Delta Sarcoglycan in Dystrophin-Glycoprotein Complex Function in Cardiac Muscle
    The Role of Delta Sarcoglycan in Dystrophin-Glycoprotein Complex Function in Cardiac Muscle by Matthew D. Campbell A dissertation submitted in partial fulfillment Of the requirements for the degree of Doctor of Philosophy (Molecular and Integrative Physiology) in The University of Michigan 2013 Doctoral Committee: Associate Professor Daniel E. Michele, Chair Professor Mark W. Russell Associate Professor Yangzhuang Wang Associate Professor Margaret V. Westfall To Shelby my strength, my life, my love ! ii! Acknowledgements The work contained in this dissertation was made possible due to the tireless efforts and support by Dan Michele. He took me into his lab when I had very little first-hand scientific experience and tried to foster the growth of an independent scientist. He supported me through all my struggles, both professional and personal. There is no way to thank him enough for what he has done for me and for pushing me beyond what I ever thought myself capable. I would like to thank my committee members Margaret, Wang, and Mark for their input and critiques. I have been blessed to work in a lab that is filled with outstanding scientists and wonderful people. I want to thank Zhyldyz, Joel, and Jessica for all the scientific input they have given, but also for being friends on whom I may lean. Former members Abbie, Sonya, and Anoop all made work much easier and helped ease all the headaches that I created. Many members of Molecular & Integrative Physiology have also been great assets especially current and former graduate chairs Fred Karsch, Ormond MacDougald, and Scott Pletcher.
    [Show full text]
  • Altered Α1-Syntrophin Expression in Myofibers With
    Histol Histopathol (2006) 21: 23-34 DOI: 10.14670/HH-21.23 Histology and Histopathology http://www.hh.um.es Cellular and Molecular Biology α Altered 1-syntrophin expression in myofibers with Duchenne and Fukuyama muscular dystrophies Y. Wakayama 1, M. Inoue 1, H. Kojima 1, T. Jimi 1, S. Yamashita 2, T. Kumagai 3, S. Shibuya 1, H. Hara 1 and H. Oniki 4 1Department of Neurology and 4Electron Microscope Laboratory, Showa University Fujigaoka Hospital, Yokohama, Japan, 2Department of Neurology, Kanagawa Children’s Medical Center, Yokohama, Japan and 3Department of Pediatric Neurology, Aichi Prefectural Colony, Kasugai, Japan α Summary. 1-Syntrophin, a scaffolding adapter and Introduction modular protein, is a cytoplasmic component of the dystrophin glycoprotein complex. This study Duchenne muscular dystrophy (DMD) is the most iαnvestigated immunohistochemically the expression of common form of human muscular dystrophy and is 1-syntrophin in Duchenne and Fukuyama muscular caused by mutations in the megagene encoding a large dystrophies (DMD and FCMD, respectively). Biopsied sarcolemmal protein dystrophin (Hoffman et al., 1987; muscles of five DMD, five FCMD, five normal controls Koenig et al., 1988). Syntrophins are a family of and five disease controls (three myotonic and two scaffolding proteins of the dystroαphin glycoprotein facioscapulohumeral dystrophies) weαre analyzed. complex that contains extracellαular -dγystδroglycan, the Immunoblot analysis showed that anti- 1-syntrophin transmembrane ß-dystroglycan, -, ß‚-, -, -sarcoglycan antibody had a decreased reaction in both DMD and and sarcospan, and cytoplasmic syntrαophins anγd FCMD muscle extracts. Biopsied muscle sections and dystrγobrevins (Ozawa, 2004). Syntrophins , ß1, ß2, 1 theirα serial sections were immunostained with rabbit and 2 have been characterized so far (Ahn et al., 1996; anti- 1-syntrophin and rabbit anti-muscle-specific ß- Piluso et al., 2000).
    [Show full text]
  • Evolving Approaches to Genetic Evaluation of Specific Cardiomyopathies
    Curr Heart Fail Rep DOI 10.1007/s11897-015-0271-7 BIOMARKERS OF HEART FAILURE (W H W TANG, SECTION EDITOR) Evolving Approaches to Genetic Evaluation of Specific Cardiomyopathies Loon Yee Louis Teo1 & Rocio T. Moran 2 & W. H. Wilson Tang 1,3 # Springer Science+Business Media New York 2015 Abstract The understanding of the genetic basis of cardiomy- Keywords Genetic evaluation . Cardiomyopathy . opathy has expanded significantly over the past 2 decades. Hereditary . Genetic testing . Evolving approaches The increasing availability, shortening diagnostic time, and lowering costs of genetic testing have provided researchers and physicians with the opportunity to identify the underlying Introduction genetic determinants for thousands of genetic disorders, in- cluding inherited cardiomyopathies, in effort to improve pa- In the contemporary definitions and classification of cardio- tient morbidities and mortality. As such, genetic testing has myopathies by the American Heart Association in 2006, car- advanced from basic scientific research to clinical application diomyopathies represent a heterogeneous group of diseases of and has been incorporated as part of patient evaluations for the myocardium associated with mechanical and/or electrical suspected inherited cardiomyopathies. Genetic evaluation dysfunction, and can be divided into primary cardiomyopa- framework of inherited cardiomyopathies typically encom- thies which predominantly involve the heart muscle, or sec- passes careful evaluation of family history, genetic counseling, ondary cardiomyopathies
    [Show full text]