Effects of trichostatin A on in vitro development and transgene function in somatic cell nuclear transfer embryos derived from transgenic Clawn miniature pig cells
Effects of trichostatin A on in vitro development and transgene function in somatic cell nuclear transfer embryos derived from transgenic Clawn miniature pig cells
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DOI：
10.1111/j.1740-0929.2010.00772.x
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发表时间：
2010-01-01
影响因子：
2
通讯作者：
Yoshida, Mitsutoshi
中科院分区：
文献类型：
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作者：
Himaki, Takehiro;Yokomine, Taka-aki;Yoshida, Mitsutoshi
本研究旨在研究组蛋白去乙酰化酶抑制剂trichostatin A （TSA）活化后处理对小爪猪胚胎成纤维细胞（PEF）体外发育和体细胞核移植（SCNT）胚胎的影响，该胚胎来源于细菌内切-半乳糖苷酶C基因（去除α -半乳糖（Gal）表位）。SCNT胚胎活化后分别加入或不加入TSA（50或100 nmol/L）孵育，体外培养，评估卵裂、囊胚形成和转基因功能。50 nmol/L TSA处理的SCNT胚胎囊胚形成率显著高于对照组（P < 0.05），而囊胚的卵裂率和细胞数无显著差异。用异硫氰酸荧光素标记的BS-I-B(4)隔离素标记后，转基因SCNT囊胚的细胞表面荧光强度明显低于非转基因SCNT囊胚。然而，转基因SCNT胚泡α - gal表位表达的降低不受TSA处理的影响。本研究结果表明，50 nmol/L TSA激活后处理可在不改变转基因功能的情况下，有效提高转基因SCNT微型猪胚胎的体外发育能力。
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-beta-galactosidase C gene (removal of the alpha-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 alpha-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.