DOCSLIB.ORG

Svcv) in Ontario

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Svcv) in Ontario PATHOGENESIS AND DISTRIBUTION OF SPRING VIREMIA OF CARP VIRUS (SVCV) IN ONTARIO by Ehab Misk A Thesis Presented to The University of Guelph In partial fulfillment of requirements for the degree of Doctor of Philosophy in Pathobiology Guelph, Ontario, Canada © Ehab Misk, May 2016 ABSTRACT PATHOGENESIS AND DISTRIBUTION OF SPRING VIREMIA OF CARP VIRUS (SVCV) IN ONTARIO Ehab Misk Advisor: University of Guelph, 2016 Professor John S. Lumsden Spring viremia of carp virus (SVCV), an OIE reportable rhabdovirus and fish pathogen, was identified in Canada in 2006 in Hamilton Harbour but has not been reported subsequently. SVCV can have a significant impact on cyprinids, and so the susceptibility of three baitfish species (emerald shiner Notropis atherinoides, fathead minnow Pimpheles promelas, white sucker Catostomus commersonii from the order Cypriniformes was assessed using experimental infection. Millions of baitfish are moved around the province to support the sports fishing industry, and this represents a risk for spread of the virus. Emerald shiner, fathead minnow and koi (43, 53, and 33% mortality, respectively) were highly susceptible, but white sucker or rainbow trout (12.5 and 0% mortality), were not. Infection was confirmed by virus isolation and RT-qPCR and SVCV was immunolocalized in association with histological lesions using immunohistochemistry. A one-step reverse transcription quantitative PCR (RT-qPCR) was adapted and used retrospectively to test samples collected by the Ontario Ministry of Natural Resources for surveillance from 2008 to 2012. A total of 1432 fish from 35 water bodies in Ontario were examined using RT-qPCR, but no additional fish were identified with SVCV. Finally, the pathogenesis of the Rhabdovirus-host interaction at the gill epithelium was investigated using the RTgill-W1 cells. The cell line was pretreated with UV-inactivated (killed) VHSV and recombinant FliC and then infected with viral hemorrhagic septicemia virus (VHSV). The viral load and gene expression was investigated 1, 3, and 6 d post infection (PI) with qPCR. In addition, the transcriptome response of RTgill-W1 cells at 36 h post-treatment with SVCV, VHSV, UV-inactivated VHSV and FliC were tested using microarray and RT-qPCR. Pretreatment of RTgill-W1 cells with killed VSHV induced a reduction in viral load (nucleoprotein copy number by RT-qPCR) after infection. Transcription profiles in VHSV- and SVCV-infected or killed VHSV and FliC pretreated RTgill-W1 cells 36 h post-exposure detected 24, 22, 123 and 190 differentially expressed probes, which contained several important gene candidates with a potentially key role in innate immunity to rhabdovirus infection in gill epithelium. ACKNOWLEDGEMENTS I would like to express my sincere appreciation to my advisor Dr. John Lumsden for his unlimited high standard professional academic support, career orientation, and personal support for me throughout my Ph.D. program. I would also like to extend my gratitude to my advisory committee Dr. Eva Nagy and Dr. Kyle Graver for their marvelous contribution in shaping my work in its best form. I owe a special acknowledgment to Paul Huber his professional technical support, refinement of my protocols and troubleshooting especially with the virus purification using sucrose gradients. I would also like to acknowledge to my current graduate student colleagues, Maureen Jarau, Ryan Horricks, Doran Kirkbright, Juan-Ting Liu, Paige Vroom and Jaramar Balmori-Cedeno for their support, fruitful discussions, and maintaining social interaction. I would also like to extend my gratitude to my former graduate student colleagues Dr. Lowia Al-Hussinee, Dr. Veronique Lepage, Dr. Alex Reid, Dr. Elena Contador, and Jessica Grice for providing a supportive and collaborative environment during the experimental trials portion of my thesis. I am indebted to Mr. R. Frank and M. Cornish from the Hagan Aqualab for setting up the biosecurity room for the experimental infection trials. I would also like to extend my thanks to the administrative and technical staff members of the department: Donna Kangas, Cathy Bernardi, Karla De Uslar, Marni Struyk, and Elizabeth Gilbertson, who have been very kind and helpful in their respective roles. This thesis research would not have been possible without the financial support provided by the Egyptian Government, The Ontario Ministry of Agriculture Food and Rural Affairs, the iv Ontario Ministry of Natural Resources and the National Engineering and Research Council of Canada. Finally, and most importantly I would like to thank Fatma Misk, my wife, who gave so much love, support, trust and time for me to be able to finish my studies and to my adorable daughters who make the world a better place. v DECLARATION OF WORK PERFORMED All work reported in this thesis was performed by myself under supervision of Dr. John Lumsden and my advisory committee; Dr. Eva Nagy and Dr. Kyle Garver, with following exceptions: The polyclonal antibody to SVCV and VHSV IVb used for immunohistochemistry in Chapter 2 was prepared by Paul Huber and Dr. Alex Reid. The VHSV one step RT-qPCR for VHSV and SVCV protocols used in Chapter 2, 3 and 4 was developed by Dr. Lincoln Tubbs but adapted and optimized by myself. Steve Lord from the Fish Health Laboratory, University of Guelph, performed the SVCV isolation in Chapter 2 from experimentally infected fish. The Princess Margaret Genomics Center, University Health Network, Ontario performed the labeling and processing of the microarray chips. vi TABLE OF CONTENTS 1 LITERATURE REVIEW ............................................................................................................ 1 1.1 INTRODUCTION ................................................................................................................... 1 1.2 SVC IN THE WORLD AND NORTH AMERICA ........................................................................ 3 1.3 TAXONOMY ......................................................................................................................... 4 1.4 FISH SPECIES AND LESIONS PRODUCED ............................................................................... 6 1.5 PATHOGENESIS OF THE DISEASE ......................................................................................... 8 1.5.1 Strains of the virus .......................................................................................................... 8 1.5.2 Mechanistic basis of lesion development. ....................................................................... 9 1.5.3 Innate immunity to SVCV ............................................................................................. 10 1.5.4 Transmission................................................................................................................. 12 1.5.5 Infection temperature ................................................................................................... 12 1.6 DIAGNOSIS ........................................................................................................................ 13 1.6.1 History, clinical signs, and pathological lesions .......................................................... 13 1.6.2 Virus isolation. ............................................................................................................. 15 1.6.3 Virus identification ....................................................................................................... 16 2 PATHOGENESIS OF SPRING VIREMIA OF CARP IN EMERALD SHINER NOTROPIS ATHERINOIDES RAFINESQUE, FATHEAD MINNOW PIMEPHALES PROMELAS RAFINESQUE AND WHITE SUCKER CATOSTOMUS COMMERSONII (LACEPEDE) ............................................... 22 2.1 INTRODUCTION ................................................................................................................. 24 2.2 MATERIALS AND METHODS .............................................................................................. 25 2.2.1 Virus propagation and plaque assay ............................................................................ 25 2.2.2 Fish ............................................................................................................................... 27 2.2.3 Experimental Infection and sampling ........................................................................... 27 vii 2.2.4 Total RNA Extraction ................................................................................................... 28 2.2.5 Reverse transcription polymerase chain reaction ........................................................ 29 2.2.6 Virus Isolation .............................................................................................................. 30 2.2.7 Gross pathology, histopathology, and IHC .................................................................. 30 2.2.8 Virus purification .......................................................................................................... 31 2.2.9 Rabbit antibody ............................................................................................................ 31 2.2.10 Statistical analysis .................................................................................................... 32 2.3 RESULTS ............................................................................................................................ 33 2.3.1 Experimental trial ......................................................................................................... 33 2.3.2 Detection of SVCV by RT-qPCR
Load more

Recommended publications
  • Ichthyophthirius Multifiliis As a Potential Vector of Edwardsiella
    RESEARCH LETTER Ichthyophthirius multifiliis as a potential vector of Edwardsiella ictaluri in channel catfish De-Hai Xu, Craig A. Shoemaker & Phillip H. Klesius U.S. Department of Agriculture, Agricultural Research Service, Aquatic Animal Health Research Unit, Auburn, AL, USA Correspondence: De-Hai Xu, U.S. Abstract Department of Agriculture, Agricultural Research Service, Aquatic Animal Health There is limited information on whether parasites act as vectors to transmit Research Unit, 990 Wire Road, Auburn, bacteria in fish. In this trial, we used Ichthyophthirius multifiliis and fluorescent AL 36832, USA. Tel.: +1 334 887 3741; Edwardsiella ictaluri as a model to study the interaction between parasite, bac- fax: +1 334 887 2983; terium, and fish. The percentage (23–39%) of theronts fluorescing after expo- e-mail: [email protected] sure to E. ictaluri was significantly higher than control theronts (~ 6%) using À flow cytometry. Theronts exposed to E. ictaluri at 4 9 107 CFU mL 1 showed Received 4 January 2012; accepted 30 ~ January 2012. a higher percentage ( 60%) of fluorescent theronts compared to those (42%) 9 3 À1 Final version published online 23 February exposed to 4 10 CFU mL at 4 h. All tomonts (100%) carried the bacte- 2012. rium after exposure to E. ictaluri. Edwardsiella ictaluri survived and replicated during tomont division. Confocal microscopy demonstrated that E. ictaluri was DOI: 10.1111/j.1574-6968.2012.02518.x associated with the tomont surface. Among theronts released from tomonts exposed to E. ictaluri,31–66% were observed with attached E. ictaluri. Sixty À Editor: Jeff Cole percent of fish exposed to theronts treated with 5 9 107 E.
    [Show full text]
  • FIELD GUIDE to WARMWATER FISH DISEASES in CENTRAL and EASTERN EUROPE, the CAUCASUS and CENTRAL ASIA Cover Photographs: Courtesy of Kálmán Molnár and Csaba Székely
    SEC/C1182 (En) FAO Fisheries and Aquaculture Circular I SSN 2070-6065 FIELD GUIDE TO WARMWATER FISH DISEASES IN CENTRAL AND EASTERN EUROPE, THE CAUCASUS AND CENTRAL ASIA Cover photographs: Courtesy of Kálmán Molnár and Csaba Székely. FAO Fisheries and Aquaculture Circular No. 1182 SEC/C1182 (En) FIELD GUIDE TO WARMWATER FISH DISEASES IN CENTRAL AND EASTERN EUROPE, THE CAUCASUS AND CENTRAL ASIA By Kálmán Molnár1, Csaba Székely1 and Mária Láng2 1Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary 2 National Food Chain Safety Office – Veterinary Diagnostic Directorate, Budapest, Hungary FOOD AND AGRICULTURE ORGANIZATION OF THE UNITED NATIONS Ankara, 2019 Required citation: Molnár, K., Székely, C. and Láng, M. 2019. Field guide to the control of warmwater fish diseases in Central and Eastern Europe, the Caucasus and Central Asia. FAO Fisheries and Aquaculture Circular No.1182. Ankara, FAO. 124 pp. Licence: CC BY-NC-SA 3.0 IGO The designations employed and the presentation of material in this information product do not imply the expression of any opinion whatsoever on the part of the Food and Agriculture Organization of the United Nations (FAO) concerning the legal or development status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. The mention of specific companies or products of manufacturers, whether or not these have been patented, does not imply that these have been endorsed or recommended by FAO in preference to others of a similar nature that are not mentioned. The views expressed in this information product are those of the author(s) and do not necessarily reflect the views or policies of FAO.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • A Multistep Bioinformatic Approach Detects Putative Regulatory
    BMC Bioinformatics BioMed Central Research article Open Access A multistep bioinformatic approach detects putative regulatory elements in gene promoters Stefania Bortoluzzi1, Alessandro Coppe1, Andrea Bisognin1, Cinzia Pizzi2 and Gian Antonio Danieli*1 Address: 1Department of Biology, University of Padova – Via Bassi 58/B, 35131, Padova, Italy and 2Department of Information Engineering, University of Padova – Via Gradenigo 6/B, 35131, Padova, Italy Email: Stefania Bortoluzzi - [email protected]; Alessandro Coppe - [email protected]; Andrea Bisognin - [email protected]; Cinzia Pizzi - [email protected]; Gian Antonio Danieli* - [email protected] * Corresponding author Published: 18 May 2005 Received: 12 November 2004 Accepted: 18 May 2005 BMC Bioinformatics 2005, 6:121 doi:10.1186/1471-2105-6-121 This article is available from: http://www.biomedcentral.com/1471-2105/6/121 © 2005 Bortoluzzi et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: Searching for approximate patterns in large promoter sequences frequently produces an exceedingly high numbers of results. Our aim was to exploit biological knowledge for definition of a sheltered search space and of appropriate search parameters, in order to develop a method for identification of a tractable number of sequence motifs. Results: Novel software (COOP) was developed for extraction of sequence motifs, based on clustering of exact or approximate patterns according to the frequency of their overlapping occurrences.
    [Show full text]
  • ELOVL4 Gene ELOVL Fatty Acid Elongase 4
    ELOVL4 gene ELOVL fatty acid elongase 4 Normal Function The ELOVL4 gene provides instructions for making a protein that is found primarily in the retina, the specialized light-sensitive tissue that lines the back of the eye. Within the retina, the ELOVL4 protein is produced in specialized light receptor cells ( photoreceptors). The ELOVL4 protein is also found in the brain and skin, but less is known about its activity (expression) in these structures. Inside photoreceptor cells, this protein is located in a cell structure called the endoplasmic reticulum that is involved in protein production, processing, and transport. The ELOVL4 protein plays a role in making a group of fats called very long-chain fatty acids. The protein helps add carbon molecules to long-chain fatty acids, making them very long-chain fatty acids. The function of the very long-chain fatty acids produced by the ELOVL4 protein is unknown. Health Conditions Related to Genetic Changes Stargardt macular degeneration At least three mutations in the ELOVL4 gene have been found to cause Stargardt macular degeneration. These mutations create a premature stop signal in the instructions used to make the ELOVL4 protein. As a result, the protein cannot be retained in the endoplasmic reticulum of photoreceptor cells. Instead, the ELOVL4 protein forms clumps (aggregates). These aggregates cannot make very long-chain fatty acids and may interfere with cell functions, ultimately leading to cell death. The loss of photoreceptor cells causes progressive vision loss in people with Stargardt
    [Show full text]
  • Review and Meta-Analysis of the Environmental Biology and Potential Invasiveness of a Poorly-Studied Cyprinid, the Ide Leuciscus Idus
    REVIEWS IN FISHERIES SCIENCE & AQUACULTURE https://doi.org/10.1080/23308249.2020.1822280 REVIEW Review and Meta-Analysis of the Environmental Biology and Potential Invasiveness of a Poorly-Studied Cyprinid, the Ide Leuciscus idus Mehis Rohtlaa,b, Lorenzo Vilizzic, Vladimır Kovacd, David Almeidae, Bernice Brewsterf, J. Robert Brittong, Łukasz Głowackic, Michael J. Godardh,i, Ruth Kirkf, Sarah Nienhuisj, Karin H. Olssonh,k, Jan Simonsenl, Michał E. Skora m, Saulius Stakenas_ n, Ali Serhan Tarkanc,o, Nildeniz Topo, Hugo Verreyckenp, Grzegorz ZieRbac, and Gordon H. Coppc,h,q aEstonian Marine Institute, University of Tartu, Tartu, Estonia; bInstitute of Marine Research, Austevoll Research Station, Storebø, Norway; cDepartment of Ecology and Vertebrate Zoology, Faculty of Biology and Environmental Protection, University of Lodz, Łod z, Poland; dDepartment of Ecology, Faculty of Natural Sciences, Comenius University, Bratislava, Slovakia; eDepartment of Basic Medical Sciences, USP-CEU University, Madrid, Spain; fMolecular Parasitology Laboratory, School of Life Sciences, Pharmacy and Chemistry, Kingston University, Kingston-upon-Thames, Surrey, UK; gDepartment of Life and Environmental Sciences, Bournemouth University, Dorset, UK; hCentre for Environment, Fisheries & Aquaculture Science, Lowestoft, Suffolk, UK; iAECOM, Kitchener, Ontario, Canada; jOntario Ministry of Natural Resources and Forestry, Peterborough, Ontario, Canada; kDepartment of Zoology, Tel Aviv University and Inter-University Institute for Marine Sciences in Eilat, Tel Aviv,
    [Show full text]
  • Renal Cell Neoplasms Contain Shared Tumor Type–Specific Copy Number Variations
    The American Journal of Pathology, Vol. 180, No. 6, June 2012 Copyright © 2012 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.ajpath.2012.01.044 Tumorigenesis and Neoplastic Progression Renal Cell Neoplasms Contain Shared Tumor Type–Specific Copy Number Variations John M. Krill-Burger,* Maureen A. Lyons,*† The annual incidence of renal cell carcinoma (RCC) has Lori A. Kelly,*† Christin M. Sciulli,*† increased steadily in the United States for the past three Patricia Petrosko,*† Uma R. Chandran,†‡ decades, with approximately 58,000 new cases diag- 1,2 Michael D. Kubal,§ Sheldon I. Bastacky,*† nosed in 2010, representing 3% of all malignancies. Anil V. Parwani,*†‡ Rajiv Dhir,*†‡ and Treatment of RCC is complicated by the fact that it is not a single disease but composes multiple tumor types with William A. LaFramboise*†‡ different morphological characteristics, clinical courses, From the Departments of Pathology* and Biomedical and outcomes (ie, clear-cell carcinoma, 82% of RCC ‡ Informatics, University of Pittsburgh, Pittsburgh, Pennsylvania; cases; type 1 or 2 papillary tumors, 11% of RCC cases; † the University of Pittsburgh Cancer Institute, Pittsburgh, chromophobe tumors, 5% of RCC cases; and collecting § Pennsylvania; and Life Technologies, Carlsbad, California duct carcinoma, approximately 1% of RCC cases).2,3 Benign renal neoplasms are subdivided into papillary adenoma, renal oncocytoma, and metanephric ade- Copy number variant (CNV) analysis was performed on noma.2,3 Treatment of RCC often involves surgical resec- renal cell carcinoma (RCC) specimens (chromophobe, tion of a large renal tissue component or removal of the clear cell, oncocytoma, papillary type 1, and papillary entire affected kidney because of the relatively large size of type 2) using high-resolution arrays (1.85 million renal tumors on discovery and the availability of a life-sus- probes).
    [Show full text]
  • Downloaded from [266]
    Patterns of DNA methylation on the human X chromosome and use in analyzing X-chromosome inactivation by Allison Marie Cotton B.Sc., The University of Guelph, 2005 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in The Faculty of Graduate Studies (Medical Genetics) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) January 2012 © Allison Marie Cotton, 2012 Abstract The process of X-chromosome inactivation achieves dosage compensation between mammalian males and females. In females one X chromosome is transcriptionally silenced through a variety of epigenetic modifications including DNA methylation. Most X-linked genes are subject to X-chromosome inactivation and only expressed from the active X chromosome. On the inactive X chromosome, the CpG island promoters of genes subject to X-chromosome inactivation are methylated in their promoter regions, while genes which escape from X- chromosome inactivation have unmethylated CpG island promoters on both the active and inactive X chromosomes. The first objective of this thesis was to determine if the DNA methylation of CpG island promoters could be used to accurately predict X chromosome inactivation status. The second objective was to use DNA methylation to predict X-chromosome inactivation status in a variety of tissues. A comparison of blood, muscle, kidney and neural tissues revealed tissue-specific X-chromosome inactivation, in which 12% of genes escaped from X-chromosome inactivation in some, but not all, tissues. X-linked DNA methylation analysis of placental tissues predicted four times higher escape from X-chromosome inactivation than in any other tissue. Despite the hypomethylation of repetitive elements on both the X chromosome and the autosomes, no changes were detected in the frequency or intensity of placental Cot-1 holes.
    [Show full text]
  • Cellular and Molecular Signatures in the Disease Tissue of Early
    Cellular and Molecular Signatures in the Disease Tissue of Early Rheumatoid Arthritis Stratify Clinical Response to csDMARD-Therapy and Predict Radiographic Progression Frances Humby1,* Myles Lewis1,* Nandhini Ramamoorthi2, Jason Hackney3, Michael Barnes1, Michele Bombardieri1, Francesca Setiadi2, Stephen Kelly1, Fabiola Bene1, Maria di Cicco1, Sudeh Riahi1, Vidalba Rocher-Ros1, Nora Ng1, Ilias Lazorou1, Rebecca E. Hands1, Desiree van der Heijde4, Robert Landewé5, Annette van der Helm-van Mil4, Alberto Cauli6, Iain B. McInnes7, Christopher D. Buckley8, Ernest Choy9, Peter Taylor10, Michael J. Townsend2 & Costantino Pitzalis1 1Centre for Experimental Medicine and Rheumatology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK. Departments of 2Biomarker Discovery OMNI, 3Bioinformatics and Computational Biology, Genentech Research and Early Development, South San Francisco, California 94080 USA 4Department of Rheumatology, Leiden University Medical Center, The Netherlands 5Department of Clinical Immunology & Rheumatology, Amsterdam Rheumatology & Immunology Center, Amsterdam, The Netherlands 6Rheumatology Unit, Department of Medical Sciences, Policlinico of the University of Cagliari, Cagliari, Italy 7Institute of Infection, Immunity and Inflammation, University of Glasgow, Glasgow G12 8TA, UK 8Rheumatology Research Group, Institute of Inflammation and Ageing (IIA), University of Birmingham, Birmingham B15 2WB, UK 9Institute of
    [Show full text]
  • A Novel Gene for Autosomal Dominant Stargardt-Like Macular Dystrophy with Homology to the SUR4 Protein Family
    A Novel Gene for Autosomal Dominant Stargardt-like Macular Dystrophy with Homology to the SUR4 Protein Family Albert O. Edwards,1 Larry A. Donoso,2 and Robert Ritter, III1 PURPOSE. To describe a novel gene causing a Stargardt-like fatty acid biosynthesis in the pathogenesis of macular degen- phenotype in a family with dominant macular dystrophy and eration. The PCR-based assay for the 5-bp deletion will facilitate the exclusion of all known genes within the disease locus. more accurate genetic counseling and identification of other METHODS. Meiotic breakpoint mapping in a family of 2314 branches of the family. (Invest Ophthalmol Vis Sci. 2001;42: individuals enabled refinement of the location of the disease 2652–2663) gene. The genomic organization and expression profile of known and putative genes within the critical region were determined using bioinformatics, cDNA cloning, and RT-PCR. acular dystrophies with subretinal flecks may arise from The coding sequence of genes expressed within the retina was Mmutations at multiple disease loci,1,2 the most common scanned for mutations, by using DNA sequencing. of which is the ABCA4 gene on chromosome 1 that gives rise 3,4 RESULTS. The disease-causing gene (STGD3) was further local- to Stargardt disease (STGD1) or fundus flavimaculatus. We 2,5 ized to 562 kb on chromosome 6 between D6S460 and a new previously reported a founder effect for a dominant condi- polymorphic marker centromeric to D6S1707. Of the four tion (STGD3) phenotypically similar to Stargardt disease local- genes identified within this region, all were expressed in the ized to chromosome 6q.2,6 A genealogical and molecular inves- retina or retinal pigment epithelium.
    [Show full text]
  • Supplementary Figure 1
    Zurich Open Repository and Archive University of Zurich Main Library Strickhofstrasse 39 CH-8057 Zurich www.zora.uzh.ch Year: 2012 Mutations in ABCD4 cause a new inborn error of vitamin B(12) metabolism Coelho, David ; Kim, Jaeseung C ; Miousse, Isabelle R ; Fung, Stephen ; du Moulin, Marcel ; Buers, Insa ; Suormala, Terttu ; Burda, Patricie ; Frapolli, Michele ; Stucki, Martin ; Nürnberg, Peter ; Thiele, Holger ; Robenek, Horst ; Höhne, Wolfgang ; Longo, Nicola ; Pasquali, Marzia ; Mengel, Eugen ; Watkins, David ; Shoubridge, Eric A ; Majewski, Jacek ; Rosenblatt, David S ; Fowler, Brian ; Rutsch, Frank ; Baumgartner, Matthias R Abstract: Inherited disorders of vitamin B(12) (cobalamin) have provided important clues to how this vitamin, which is essential for hematological and neurological function, is transported and metabolized. We describe a new disease that results in failure to release vitamin B(12) from lysosomes, which mimics the cblF defect caused by LMBRD1 mutations. Using microcell-mediated chromosome transfer and exome sequencing, we identified causal mutations in ABCD4, a gene that codes for an ABC transporter, which was previously thought to have peroxisomal localization and function. Our results show that ABCD4 colocalizes with the lysosomal proteins LAMP1 and LMBD1, the latter of which is deficient in the cblF defect. Furthermore, we show that mutations altering the putative ATPase domain of ABCD4 affect its function, suggesting that the ATPase activity of ABCD4 may be involved in intracellular processing of vitamin B(12).
    [Show full text]
  • Germline Mutations Causing Familial Lung Cancer
    Journal of Human Genetics (2015) 60, 597–603 & 2015 The Japan Society of Human Genetics All rights reserved 1434-5161/15 www.nature.com/jhg ORIGINAL ARTICLE Germline mutations causing familial lung cancer Koichi Tomoshige1,2, Keitaro Matsumoto1, Tomoshi Tsuchiya1, Masahiro Oikawa1, Takuro Miyazaki1, Naoya Yamasaki1, Hiroyuki Mishima2, Akira Kinoshita2, Toru Kubo3, Kiyoyasu Fukushima3, Koh-ichiro Yoshiura2 and Takeshi Nagayasu1 Genetic factors are important in lung cancer, but as most lung cancers are sporadic, little is known about inherited genetic factors. We identified a three-generation family with suspected autosomal dominant inherited lung cancer susceptibility. Sixteen individuals in the family had lung cancer. To identify the gene(s) that cause lung cancer in this pedigree, we extracted DNA from the peripheral blood of three individuals and from the blood of one cancer-free control family member and performed whole-exome sequencing. We identified 41 alterations in 40 genes in all affected family members but not in the unaffected member. These were considered candidate mutations for familial lung cancer. Next, to identify somatic mutations and/or inherited alterations in these 40 genes among sporadic lung cancers, we performed exon target enrichment sequencing using 192 samples from sporadic lung cancer patients. We detected somatic ‘candidate’ mutations in multiple sporadic lung cancer samples; MAST1, CENPE, CACNB2 and LCT were the most promising candidate genes. In addition, the MAST1 gene was located in a putative cancer-linked locus in the pedigree. Our data suggest that several genes act as oncogenic drivers in this family, and that MAST1 is most likely to cause lung cancer.
    [Show full text]