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Effects of trichostatin A on in vitro development and transgene function in somatic cell nuclear transfer embryos derived from transgenic Clawn miniature pig cells
Authors:Takehiro HIMAKI  Taka‐aki YOKOMINE  Masahiro SATO  Sonshin TAKAO  Kazuchika MIYOSHI  Mitsutoshi YOSHIDA
Institution:1. Laboratory of Animal Reproduction, The United Graduate School of Agricultural Sciences,;2. Laboratory of Frontier Medicine, and;3. Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Kagoshima, Japan
Abstract:The present study was carried out to examine the effects of post‐activation treatment of trichostatin A (TSA), a histone deacetylase inhibitor, on in vitro development and transgene function of somatic cell nuclear transfer (SCNT) embryos derived from Clawn miniature pig embryonic fibroblast (PEF) transfected with a bacterial endo‐β‐galactosidase C gene (removal of the α‐galactosyl (Gal) epitope). SCNT embryos were incubated with or without TSA (50 or 100 nmol/L) after activation, cultured in vitro and assessed for cleavage, blastocyst formation and transgene function. The rate of blastocyst formation was significantly higher in SCNT embryos treated with 50 nmol/L TSA than that in control (P < 0.05), whereas the rate of cleavage and cell number of blastocyst did not differ. Following labelling with fluorescein isothiocyanate‐labelled BS‐I‐B4 isolectin, the intensity of fluorescence observed on cell‐surface was dramatically reduced in transgenic SCNT blastocyst in comparison with non‐transgenic SCNT blastocyst. However, the reduction of α‐Gal epitope expression in transgenic SCNT blastocyst was not affected by TSA treatment. The results of this study showed that post‐activation treatment with 50 nmol/L TSA is effective to improve in vitro developmental capacity of transgenic SCNT miniature pig embryos without the modification of transgene function.
Keywords:Clawn miniature pig  endo‐β  ‐galactosidase C  gene‐modification  somatic cell nuclear transfer  trichostatin A
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