title: Mutated Huntingtin Causes Testicular Pathology in Transgenic Minipig Boars.
abstract: BACKGROUND
  Huntington's disease is induced by CAG expansion in a single gene coding the huntingtin protein. The mutated huntingtin (mtHtt) primarily causes degeneration of neurons in the brain, but it also affects peripheral tissues, including testes.
  
  
  OBJECTIVE
  We studied sperm and testes of transgenic boars expressing the N-terminal region of human mtHtt.
  
  
  METHODS
  In this study, measures of reproductive parameters and electron microscopy (EM) images of spermatozoa and testes of transgenic (TgHD) and wild-type (WT) boars of F1 (24-48 months old) and F2 (12-36 months old) generations were compared. In addition, immunofluorescence, immunohistochemistry, Western blot, hormonal analysis and whole-genome sequencing were done in order to elucidate the effects of mtHtt.
  
  
  RESULTS
  Evidence for fertility failure of both TgHD generations was observed at the age of 13 months. Reproductive parameters declined and progressively worsened with age. EM revealed numerous pathological features in sperm tails and in testicular epithelium from 24- and 36-month-old TgHD boars. Moreover, immunohistochemistry confirmed significantly lower proliferation activity of spermatogonia in transgenic testes. mtHtt was highly expressed in spermatozoa and testes of TgHD boars and localized in all cells of seminiferous tubules. Levels of fertility-related hormones did not differ in TgHD and WT siblings. Genome analysis confirmed that insertion of the lentiviral construct did not interrupt any coding sequence in the pig genome.
  
  
  CONCLUSIONS
  The sperm and testicular degeneration of TgHD boars is caused by gain-of-function of the highly expressed mtHtt.
authors.affiliation: Laboratory of Cell Regeneration and Plasticity, Institute of Animal Physiology and Genetics, Czech Academy of Science, Libechov, Czech Republic.
authors.forename: Monika
authors.initials: M
authors.lastname: Macakova
authors.forename: Bozena
authors.initials: B
authors.lastname: Bohuslavova
authors.forename: Petra
authors.initials: P
authors.lastname: Vochozkova
authors.forename: Antonin
authors.initials: A
authors.lastname: Pavlok
authors.forename: Miroslava
authors.initials: M
authors.lastname: Sedlackova
authors.forename: Daniela
authors.initials: D
authors.lastname: Vidinska
authors.forename: Klara
authors.initials: K
authors.lastname: Vochyanova
authors.forename: Irena
authors.initials: I
authors.lastname: Liskova
authors.forename: Ivona
authors.initials: I
authors.lastname: Valekova
authors.forename: Monika
authors.initials: M
authors.lastname: Baxa
authors.forename: Zdenka
authors.initials: Z
authors.lastname: Ellederova
authors.forename: Jiri
authors.initials: J
authors.lastname: Klima
authors.forename: Stefan
authors.initials: S
authors.lastname: Juhas
authors.forename: Jana
authors.initials: J
authors.lastname: Juhasova
authors.forename: Jana
authors.initials: J
authors.lastname: Klouckova
authors.forename: Martin
authors.initials: M
authors.lastname: Haluzik
authors.forename: Jiri
authors.initials: J
authors.lastname: Klempir
authors.forename: Hana
authors.initials: H
authors.lastname: Hansikova
authors.forename: Jana
authors.initials: J
authors.lastname: Spacilova
authors.forename: Ryan
authors.initials: R
authors.lastname: Collins
authors.forename: Ian
authors.initials: I
authors.lastname: Blumenthal
authors.forename: Michael
authors.initials: M
authors.lastname: Talkowski
authors.forename: James F
authors.initials: JF
authors.lastname: Gusella
authors.forename: David S
authors.initials: DS
authors.lastname: Howland
authors.forename: Marian
authors.initials: M
authors.lastname: DiFiglia
authors.forename: Jan
authors.initials: J
authors.lastname: Motlik
mesh_terms: D000375:Aging / Q000378:metabolism / Q000473:pathology; D000818:Animals; D030801:Animals, Genetically Modified; D049109:Cell Proliferation / Q000502:physiology; D004195:Disease Models, Animal; D005822:Genetic Vectors; D006801:Humans; D000071058:Huntingtin Protein / Q000235:genetics / Q000378:metabolism*; D006816:Huntington Disease / Q000378:metabolism / Q000473:pathology; D016086:Lentivirus / Q000235:genetics; D008297:Male; D009154:Mutation; D013076:Sperm Count; D013094:Spermatozoa / Q000378:metabolism* / Q000473:pathology*; D013552:Swine; D013556:Swine, Miniature; D013737:Testis / Q000378:metabolism* / Q000473:pathology*
chemical_list: C086055:HTT protein, human; D000071058:Huntingtin Protein