Vichera G, Alfonso J, Duque C C, Silvestre M A, Pereyra-Bonnet F, Fernández-Martín R, Salamone D
Laboratorio de Biotecnología Animal, Facultad de Agronomía, Universidad de Buenos Aires, Buenos Aires, Argentina.
Reprod Domest Anim. 2010 Dec;45(6):e306-12. doi: 10.1111/j.1439-0531.2009.01563.x.
The aim of this study was to evaluate the potential of dehydroleucodine (DhL), a new drug isolated from a medicinal herb used in Argentina, for activation of bovine oocyte. Several DhL concentrations and exposure times after ionomycin (Io) treatment were tested. The optimal DhL treatment, found for parthenogenetic development, was employed to produce bovine embryos by intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT). The best parthenogenic embryo developments were observed with 5 μM Io for 4 min followed by 5 μM DhL concentration and after 3-h exposure time (52.3% cleavage; 17.4% morulae; 7.3% blastocyst; n = 109). This treatment generated no significant differences with standard Io plus 6-dimethylaminopurine (DMAP) treatment in preimplantation embryo development. In our conditions, the embryo development reached after ICSI and SCNT assisted by the DhL treatment did not differ in terms of cleavage and blastocyst development from activation with standard Io plus DMAP treatment (p > 0.05). In conclusion, DhL utilization to activate oocytes and induce development of parthenogenotes, ICSI-embryos or SCNT-embryos is reported here for first time.
本研究的目的是评估从阿根廷一种药用植物中分离出的新药脱氢白屈菜碱（DhL）激活牛卵母细胞的潜力。测试了离子霉素（Io）处理后几种DhL浓度和暴露时间。用于孤雌生殖发育的最佳DhL处理方法被用于通过胞浆内精子注射（ICSI）和体细胞核移植（SCNT）生产牛胚胎。在5 μM Io处理4分钟，随后5 μM DhL浓度并暴露3小时后观察到最佳孤雌胚胎发育（52.3%卵裂；17.4%桑葚胚；7.3%囊胚；n = 109）。在植入前胚胎发育方面，这种处理与标准Io加6 - 二甲基氨基嘌呤（DMAP）处理没有显著差异。在我们的条件下，DhL处理辅助的ICSI和SCNT后达到的胚胎发育在卵裂和囊胚发育方面与标准Io加DMAP处理激活没有差异（p>0.05）。总之，本文首次报道了利用DhL激活卵母细胞并诱导孤雌胚胎、ICSI胚胎或SCNT胚胎发育。
