Data di Pubblicazione:
2013
Citazione:
A transgenic minipig models of Huntington’s disease / M. Baxa, M. Hruska Plochan, S. Juhas, P. Vodicka, A. Pavlok, J. Juhasova, A. Miyanohara, T. Nejime, J. Klima, M. Macakova, S. Marsala, A. Weiss, S. Kubickova, P. Musilova, R. Vrtel, E.M. Sontag, L.M. Thompson, J. Schier, H. Hansikova, D.S. Howland, E. Cattaneo, M. Difiglia, M. Marsala, J. Motlik. - In: JOURNAL OF HUNTINGTON’S DISEASE. - ISSN 1879-6397. - 2:1(2013), pp. 47-78. [10.3233/JHD-130001]
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 HTTmRNAand 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 siblings 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.
Tipologia IRIS:
01 - Articolo su periodico
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
Elenco autori:
M. Baxa, M. Hruska Plochan, S. Juhas, P. Vodicka, A. Pavlok, J. Juhasova, A. Miyanohara, T. Nejime, J. Klima, M. Macakova, S. Marsala, A. Weiss, S. Kubickova, P. Musilova, R. Vrtel, E.M. Sontag, L.M. Thompson, J. Schier, H. Hansikova, D.S. Howland, E. Cattaneo, M. Difiglia, M. Marsala, J. Motlik
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