Effects of trichostatin A on in vitro development and transgene function in somatic cell nuclear transfer embryos derived from transgenic Clawn miniature pig cells.
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-B(4) 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.
Himaki
Takehiro
T
Laboratory of Animal Reproduction, The United Graduate School of Agricultural Sciences, Frontier Science Research Center, Kagoshima University, Kagoshima, Japan.
Yokomine
Taka-Aki
TA
Sato
Masahiro
M
Takao
Sonshin
S
Miyoshi
Kazuchika
K
Yoshida
Mitsutoshi
M
D000818:Animals; D004622:Embryo, Mammalian / Q000187:drug effects; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006877:Hydroxamic Acids / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D013552:Swine; D013556:Swine, Miniature / Q000196:embryology*; D019076:Transgenes / Q000187:drug effects*
D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; C012589:trichostatin A