title: A transgenic minipig model of Huntington's Disease.
abstract: BACKGROUND
  Some promising treatments for Huntington's disease (HD) may require pre-clinical testing in large animals. Minipig is a suitable species because of its large gyrencephalic brain and long lifespan.
  
  
  OBJECTIVE
  To generate HD transgenic (TgHD) minipigs encoding huntingtin (HTT)1-548 under the control of human HTT promoter.
  
  
  METHODS
  Transgenesis was achieved by lentiviral infection of porcine embryos. PCR assessment of gene transfer, observations of behavior, and postmortem biochemical and immunohistochemical studies were conducted.
  
  
  RESULTS
  One copy of the human HTT transgene encoding 124 glutamines integrated into chromosome 1 q24-q25 and successful germ line transmission occurred through successive generations (F0, F1, F2 and F3 generations). No developmental or gross motor deficits were noted up to 40 months of age. Mutant HTT mRNA and protein fragment were detected in brain and peripheral tissues. No aggregate formation in brain up to 16 months was seen by AGERA and filter retardation or by immunostaining. DARPP32 labeling in WT and TgHD minipig neostriatum was patchy. Analysis of 16 month old sibling pairs showed reduced intensity of DARPP32 immunoreactivity in neostriatal TgHD neurons compared to those of WT. Compared to WT, TgHD boars by one year had reduced fertility and fewer spermatozoa per ejaculate. In vitro analysis revealed a significant decline in the number of WT minipig oocytes penetrated by TgHD spermatozoa.
  
  
  CONCLUSIONS
  The findings demonstrate successful establishment of a transgenic model of HD in minipig that should be valuable for testing long term safety of HD therapeutics. The emergence of HD-like phenotypes in the TgHD minipigs will require more study.
authors: Laboratory of Cell Regeneration and Plasticity, Institute of Animal Physiology and Genetics, v.v.i., AS CR, Libechov, Czech Republic Faculty of Science, Department of Cell Biology, Charles University in Prague, Prague, Czech Republic.
authors: Monika
authors: M
authors: Baxa
authors: Laboratory of Cell Regeneration and Plasticity, Institute of Animal Physiology and Genetics, v.v.i., AS CR, Libechov, Czech Republic Faculty of Science, Department of Cell Biology, Charles University in Prague, Prague, Czech Republic Neurodegeneration Laboratory, Department of Anesthesiology, University of California, San Diego, La Jolla, CA, USA Sanford Consortium for Regenerative Medicine, San Diego, La Jolla, CA, USA.
authors: Marian
authors: M
authors: Hruska-Plochan
authors: Laboratory of Cell Regeneration and Plasticity, Institute of Animal Physiology and Genetics, v.v.i., AS CR, Libechov, Czech Republic.
authors: Stefan
authors: S
authors: Juhas
authors: Laboratory of Cell Regeneration and Plasticity, Institute of Animal Physiology and Genetics, v.v.i., AS CR, Libechov, Czech Republic Department of Neurology, Massachusetts General Hospital, Boston, MA, USA.
authors: Petr
authors: P
authors: Vodicka
authors: Laboratory of Cell Regeneration and Plasticity, Institute of Animal Physiology and Genetics, v.v.i., AS CR, Libechov, Czech Republic.
authors: Antonin
authors: A
authors: Pavlok
authors: Laboratory of Cell Regeneration and Plasticity, Institute of Animal Physiology and Genetics, v.v.i., AS CR, Libechov, Czech Republic.
authors: Jana
authors: J
authors: Juhasova
authors: Vector Development Laboratory, Human Gene Therapy Program, Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.
authors: Atsushi
authors: A
authors: Miyanohara
authors: Neurodegeneration Laboratory, Department of Anesthesiology, University of California, San Diego, La Jolla, CA, USA.
authors: Tetsuya
authors: T
authors: Nejime
authors: Laboratory of Cell Regeneration and Plasticity, Institute of Animal Physiology and Genetics, v.v.i., AS CR, Libechov, Czech Republic.
authors: Jiri
authors: J
authors: Klima
authors: Laboratory of Cell Regeneration and Plasticity, Institute of Animal Physiology and Genetics, v.v.i., AS CR, Libechov, Czech Republic Faculty of Science, Department of Cell Biology, Charles University in Prague, Prague, Czech Republic.
authors: Monika
authors: M
authors: Macakova
authors: Neurodegeneration Laboratory, Department of Anesthesiology, University of California, San Diego, La Jolla, CA, USA Sanford Consortium for Regenerative Medicine, San Diego, La Jolla, CA, USA.
authors: Silvia
authors: S
authors: Marsala
authors: Novartis Institutes for Biomedical Research, Neuroscience Discovery, Basel, Switzerland IRBM Promidis, Pomezia, Italy.
authors: Andreas
authors: A
authors: Weiss
authors: Department of Genetics and Reproduction, Veterinary Research Institute, Brno, Czech Republic.
authors: Svatava
authors: S
authors: Kubickova
authors: Department of Genetics and Reproduction, Veterinary Research Institute, Brno, Czech Republic.
authors: Petra
authors: P
authors: Musilova
authors: Department of Clinical Genetics and Fetal Medicine, Palacky University, University Hospital Olomouc, Olomouc, Czech Republic.
authors: Radek
authors: R
authors: Vrtel
authors: Department of Biological Chemistry University of California, Irvine, CA, USA Department of Psychiatry and Human Behavior, University of California, Irvine, CA, USA.
authors: Emily M
authors: EM
authors: Sontag
authors: Department of Biological Chemistry University of California, Irvine, CA, USA Department of Psychiatry and Human Behavior, University of California, Irvine, CA, USA Department of Neurobiology and Behavior University of California, Irvine, CA, USA.
authors: Leslie M
authors: LM
authors: Thompson
authors: Institute of Information Theory and Automation v.v.i., AS CR, Prague, Czech Republic.
authors: Jan
authors: J
authors: Schier
authors: Laboratory for Study of Mitochondrial Disorders, First Faculty of Medicine, Department of Pediatrics and Adolescent Medicine, Charles University and General University Hospital in Prague, Prague, Czech Republic.
authors: Hana
authors: H
authors: Hansikova
authors: CHDI Foundation, Princeton, NY, USA.
authors: David S
authors: DS
authors: Howland
authors: Department of Pharmacological Sciences and Centre for Stem Cell Research, Università degli Studi di Milano, Milan, Italy.
authors: Elena
authors: E
authors: Cattaneo
authors: Department of Neurology, Massachusetts General Hospital, Boston, MA, USA.
authors: Marian
authors: M
authors: DiFiglia
authors: Neurodegeneration Laboratory, Department of Anesthesiology, University of California, San Diego, La Jolla, CA, USA Sanford Consortium for Regenerative Medicine, San Diego, La Jolla, CA, USA Institute of Neurobiology, Slovak Academy of Sciences, Kosice, Slovak Republic.
authors: Martin
authors: M
authors: Marsala
authors: Laboratory of Cell Regeneration and Plasticity, Institute of Animal Physiology and Genetics, v.v.i., AS CR, Libechov, Czech Republic.
authors: Jan
authors: J
authors: Motlik
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D015153:Blotting, Western; D004195:Disease Models, Animal; D005260:Female; D005822:Genetic Vectors; D000071058:Huntingtin Protein; D006816:Huntington Disease; D017403:In Situ Hybridization; D016086:Lentivirus; D008297:Male; D009419:Nerve Tissue Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013552:Swine; D013556:Swine, Miniature; D014161:Transduction, Genetic; D019076:Transgenes
chemical_list: C086055:HTT protein, human; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins
keywords: AGERA assay; DARPP32; FISH analysis; Huntington's disease; TR-FRET assay; immunohistochemistry; large animal model; lentiviral transgenesis; mRNA and protein expression; minipigs; mutant huntingtin; spermatozoa