Himaki Takehiro, Mizobe Yamato, Tsuda Kenichirou, Suetomo Masashi, Yamakuchi Hiroshi, Miyoshi Kazuchika, Takao Sonshin, Yoshida Mitsutoshi
Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University, Kagoshima 890-0065, Japan.
J Reprod Dev. 2012;58(4):398-403. doi: 10.1262/jrd.11-083a. Epub 2012 Apr 13.
The objective of this study was to examine the effect of postactivation treatment with latrunculin A (LatA), an actin polymerization inhibitor, on in vitro and in vivo development of somatic cell nuclear transfer (SCNT) embryos derived from kidney fibroblasts of an aged Clawn miniature boar (12 years old). After electric activation, SCNT embryos were treated with 0, 0.5 or 1 μM LatA and cultured in vitro. The rate of blastocyst formation was significantly higher (P<0.05) in SCNT embryos treated with 0.5 μM LatA (38%) than those in control (14%). When cloned embryos treated with 0.5 μM LatA were transferred into the oviducts of two recipient miniature gilts to assess their development in vivo, both recipients became pregnant; one maintained pregnancy to term, and a live piglet (weighing 220 g) was delivered by Caesarean section. The results of this study indicated that the postactivation treatment with LatA was effective in improving in vitro developmental capacity of SCNT miniature pig embryos derived from kidney fibroblasts of an aged animal and that miniature pig cloned embryos treated with LatA had the ability to develop to term.
本研究的目的是检测用肌动蛋白聚合抑制剂拉春库林A(LatA)进行激活后处理,对来自一头12岁高龄的克劳恩小型猪肾脏成纤维细胞的体细胞核移植(SCNT)胚胎体外和体内发育的影响。电激活后,SCNT胚胎用0、0.5或1 μM的LatA处理并进行体外培养。用0.5 μM LatA处理的SCNT胚胎的囊胚形成率(38%)显著高于对照组(14%)(P<0.05)。当用0.5 μM LatA处理的克隆胚胎被移植到两头受体小型后备母猪的输卵管中以评估其体内发育情况时,两头受体均怀孕;其中一头维持妊娠至足月,并通过剖腹产分娩出一头活仔猪(体重220 g)。本研究结果表明,用LatA进行激活后处理可有效提高来自老龄动物肾脏成纤维细胞的SCNT小型猪胚胎的体外发育能力,且经LatA处理的小型猪克隆胚胎具有发育至足月的能力。